WorldWideScience

Sample records for act intravital multiphoton

  1. Video-rate resonant scanning multiphoton microscopy: An emerging technique for intravital imaging of the tumor microenvironment

    OpenAIRE

    Kirkpatrick, Nathaniel D.; Chung, Euiheon; Cook, Daniel C.; Han, Xiaoxing; Gruionu, Gabriel; Liao, Shan; Munn, Lance L.; Padera, Timothy P.; Fukumura, Dai; Jain, Rakesh K.

    2012-01-01

    The abnormal tumor microenvironment fuels tumor progression, metastasis, immune suppression, and treatment resistance. Over last several decades, developments in and applications of intravital microscopy have provided unprecedented insights into the dynamics of the tumor microenvironment. In particular, intravital multiphoton microscopy has revealed the abnormal structure and function of tumor-associated blood and lymphatic vessels, the role of aberrant tumor matrix in drug delivery, invasion...

  2. Usefulness of Intravital Multiphoton Microscopy in Visualizing Study of Mouse Cochlea and Volume Changes in the Scala Media.

    Science.gov (United States)

    Ju, Hyun Mi; Lee, Sun Hee; Kong, Tae Hoon; Kwon, Seung-Hae; Choi, Jin Sil; Seo, Young Joon

    2017-01-01

    Conventional microscopy has limitations in viewing the cochlear microstructures due to three-dimensional spiral structure and the overlying bone. But these issues can be overcome by imaging the cochlea in vitro with intravital multiphoton microscopy (MPM). By using near-infrared lasers for multiphoton excitation, intravital MPM can detect endogenous fluorescence and second harmonic generation of tissues. In this study, we used intravital MPM to visualize various cochlear microstructures without any staining and non-invasively analyze the volume changes of the scala media (SM) without removing the overlying cochlear bone. The intravital MPM images revealed various tissue types, ranging from thin membranes to dense bone, as well as the spiral ganglion beneath the cochlear bone. The two-dimensional, cross-sectional, and serial z-stack intravital MPM images also revealed the spatial dilation of the SM in the temporal bone of pendrin-deficient mice. These findings suggest that intravital MPM might serve as a new method for obtaining microanatomical information regarding the cochlea, similar to standard histopathological analyses in the animal study for the cochlea. Given the capability of intravital MPM for detecting an increase in the volume of the SM in pendrin-deficient mice, it might be a promising new tool for assessing the pathophysiology of hearing loss in the future.

  3. Usefulness of Intravital Multiphoton Microscopy in Visualizing Study of Mouse Cochlea and Volume Changes in the Scala Media

    Directory of Open Access Journals (Sweden)

    Hyun Mi Ju

    2017-07-01

    Full Text Available Conventional microscopy has limitations in viewing the cochlear microstructures due to three-dimensional spiral structure and the overlying bone. But these issues can be overcome by imaging the cochlea in vitro with intravital multiphoton microscopy (MPM. By using near-infrared lasers for multiphoton excitation, intravital MPM can detect endogenous fluorescence and second harmonic generation of tissues. In this study, we used intravital MPM to visualize various cochlear microstructures without any staining and non-invasively analyze the volume changes of the scala media (SM without removing the overlying cochlear bone. The intravital MPM images revealed various tissue types, ranging from thin membranes to dense bone, as well as the spiral ganglion beneath the cochlear bone. The two-dimensional, cross-sectional, and serial z-stack intravital MPM images also revealed the spatial dilation of the SM in the temporal bone of pendrin-deficient mice. These findings suggest that intravital MPM might serve as a new method for obtaining microanatomical information regarding the cochlea, similar to standard histopathological analyses in the animal study for the cochlea. Given the capability of intravital MPM for detecting an increase in the volume of the SM in pendrin-deficient mice, it might be a promising new tool for assessing the pathophysiology of hearing loss in the future.

  4. Video-rate resonant scanning multiphoton microscopy: An emerging technique for intravital imaging of the tumor microenvironment.

    Science.gov (United States)

    Kirkpatrick, Nathaniel D; Chung, Euiheon; Cook, Daniel C; Han, Xiaoxing; Gruionu, Gabriel; Liao, Shan; Munn, Lance L; Padera, Timothy P; Fukumura, Dai; Jain, Rakesh K

    2012-01-01

    The abnormal tumor microenvironment fuels tumor progression, metastasis, immune suppression, and treatment resistance. Over last several decades, developments in and applications of intravital microscopy have provided unprecedented insights into the dynamics of the tumor microenvironment. In particular, intravital multiphoton microscopy has revealed the abnormal structure and function of tumor-associated blood and lymphatic vessels, the role of aberrant tumor matrix in drug delivery, invasion and metastasis of tumor cells, the dynamics of immune cell trafficking to and within tumors, and gene expression in tumors. However, traditional multiphoton microscopy suffers from inherently slow imaging rates-only a few frames per second, thus unable to capture more rapid events such as blood flow, lymphatic flow, and cell movement within vessels. Here, we report the development and implementation of a video-rate multiphoton microscope (VR-MPLSM) based on resonant galvanometer mirror scanning that is capable of recording at 30 frames per second and acquiring intravital multispectral images. We show that the design of the system can be readily implemented and is adaptable to various experimental models. As examples, we demonstrate the utility of the system to directly measure flow within tumors, capture metastatic cancer cells moving within the brain vasculature and cells in lymphatic vessels, and image acute responses to changes in a vascular network. VR-MPLSM thus has the potential to further advance intravital imaging and provide new insight into the biology of the tumor microenvironment.

  5. From morphology to biochemical state - intravital multiphoton fluorescence lifetime imaging of inflamed human skin

    Science.gov (United States)

    Huck, Volker; Gorzelanny, Christian; Thomas, Kai; Getova, Valentina; Niemeyer, Verena; Zens, Katharina; Unnerstall, Tim R.; Feger, Julia S.; Fallah, Mohammad A.; Metze, Dieter; Ständer, Sonja; Luger, Thomas A.; Koenig, Karsten; Mess, Christian; Schneider, Stefan W.

    2016-03-01

    The application of multiphoton microscopy in the field of biomedical research and advanced diagnostics promises unique insights into the pathophysiology of inflammatory skin diseases. In the present study, we combined multiphoton-based intravital tomography (MPT) and fluorescence lifetime imaging (MPT-FLIM) within the scope of a clinical trial of atopic dermatitis with the aim of providing personalised data on the aetiopathology of inflammation in a non-invasive manner at patients’ bedsides. These ‘optical biopsies’ generated via MPT were morphologically analysed and aligned with classical skin histology. Because of its subcellular resolution, MPT provided evidence of a redistribution of mitochondria in keratinocytes, indicating an altered cellular metabolism. Two independent morphometric algorithms reliably showed an even distribution in healthy skin and a perinuclear accumulation in inflamed skin. Moreover, using MPT-FLIM, detection of the onset and progression of inflammatory processes could be achieved. In conclusion, the change in the distribution of mitochondria upon inflammation and the verification of an altered cellular metabolism facilitate a better understanding of inflammatory skin diseases and may permit early diagnosis and therapy.

  6. Intravital multiphoton imaging reveals multicellular streaming as a crucial component of in vivo cell migration in human breast tumors

    Science.gov (United States)

    Patsialou, Antonia; Bravo-Cordero, Jose Javier; Wang, Yarong; Entenberg, David; Liu, Huiping; Clarke, Michael; Condeelis, John S.

    2014-01-01

    Metastasis is the main cause of death in breast cancer patients. Cell migration is an essential component of almost every step of the metastatic cascade, especially the early step of invasion inside the primary tumor. In this report, we have used intravital multiphoton microscopy to visualize the different migration patterns of human breast tumor cells in live primary tumors. We used xenograft tumors of MDA-MB-231 cells as well as a low passage xenograft tumor from orthotopically injected patient-derived breast tumor cells. Direct visualization of human tumor cells in vivo shows two patterns of high-speed migration inside primary tumors: a. single cells and b. multicellular streams (i.e., cells following each other in a single file but without cohesive cell junctions). Critically, we found that only streaming and not random migration of single cells was significantly correlated with proximity to vessels, with intravasation and with numbers of elevated circulating tumor cells in the bloodstream. Finally, although the two human tumors were derived from diverse genetic backgrounds, we found that their migratory tumor cells exhibited coordinated gene expression changes that led to the same end-phenotype of enhanced migration involving activating actin polymerization and myosin contraction. Our data are the first direct visualization and assessment of in vivo migration within a live patient-derived breast xenograft tumor. PMID:25013744

  7. If you don't look, you won't see: intravital multiphoton imaging of primary and metastatic breast cancer

    NARCIS (Netherlands)

    Bonapace, L.; Wyckoff, J.; Oertner, T.; van Rheenen, J.; Junt, T.; Bentires-Alj, M.

    2012-01-01

    A fundamental hallmark of cancer is progression to metastasis and the growth of breast cancer metastases in lung, bone, liver and/or brain causes fatal complications. Unfortunately, the cellular and biochemical mechanisms of the metastatic process remain ill-defined. Recent application of intravital

  8. Correlating Intravital Multi-Photon Microscopy to 3D Electron Microscopy of Invading Tumor Cells Using Anatomical Reference Points

    Science.gov (United States)

    Karreman, Matthia A.; Mercier, Luc; Schieber, Nicole L.; Shibue, Tsukasa; Schwab, Yannick; Goetz, Jacky G.

    2014-01-01

    Correlative microscopy combines the advantages of both light and electron microscopy to enable imaging of rare and transient events at high resolution. Performing correlative microscopy in complex and bulky samples such as an entire living organism is a time-consuming and error-prone task. Here, we investigate correlative methods that rely on the use of artificial and endogenous structural features of the sample as reference points for correlating intravital fluorescence microscopy and electron microscopy. To investigate tumor cell behavior in vivo with ultrastructural accuracy, a reliable approach is needed to retrieve single tumor cells imaged deep within the tissue. For this purpose, fluorescently labeled tumor cells were subcutaneously injected into a mouse ear and imaged using two-photon-excitation microscopy. Using near-infrared branding, the position of the imaged area within the sample was labeled at the skin level, allowing for its precise recollection. Following sample preparation for electron microscopy, concerted usage of the artificial branding and anatomical landmarks enables targeting and approaching the cells of interest while serial sectioning through the specimen. We describe here three procedures showing how three-dimensional (3D) mapping of structural features in the tissue can be exploited to accurately correlate between the two imaging modalities, without having to rely on the use of artificially introduced markers of the region of interest. The methods employed here facilitate the link between intravital and nanoscale imaging of invasive tumor cells, enabling correlating function to structure in the study of tumor invasion and metastasis. PMID:25479106

  9. Intravital multiphoton tomography as a novel tool for non-invasive in vivo analysis of human skin affected with atopic dermatitis

    Science.gov (United States)

    Huck, Volker; Gorzelanny, Christian; Thomas, Kai; Niemeyer, Verena; Luger, Thomas A.; König, Karsten; Schneider, Stefan W.

    2010-02-01

    Atopic Dermatitis (AD) is an inflammatory disease of human skin. Its pathogenesis is still unknown; however, dysfunctions of the epidermal barrier and the immune response are regarded as key factors for the development of AD. In our study we applied intravital multiphoton tomography (5D-IVT), equipped with a spectral-FLIM module for in-vivo and ex-vivo analysis of human skin affected with AD. In addition to the morphologic skin analysis, FLIM technology gain access to the metabolic status of the epidermal cells referring to the NADH specific fluorescence lifetime. We evaluated a characteristic 5D-IVT skin pattern of AD in comparison to histological sections and detected a correlation with the disease activity measured by SCORAD. FLIM analysis revealed a shift of the mean fluorescence lifetime (taum) of NADH, indicating an altered metabolic activity. Within an ex-vivo approach we have investigated cryo-sections of human skin with or without barrier defects. Spectral-FLIM allows the detection of autofluorescent signals that reflect the pathophysiological conditions of the defect skin barrier. In our study the taum value was shown to be different between healthy and affected skin. Application of the 5D-IVT allows non-invasive in-vivo imaging of human skin with a penetration depth of 150 μm. We could show that affected skin could be distinguished from healthy skin by morphological criteria, by FLIM and by spectral-FLIM. Further studies will evaluate the application of the 5D-IVT technology as a diagnostic tool and to monitor the therapeutic efficacy.

  10. Intravital Imaging

    OpenAIRE

    Pittet, Mikael J.; Weissleder, Ralph

    2011-01-01

    Until recently, the idea of observing life deep within the tissues of a living mouse, at a resolution sufficient to pick out cellular behaviors and molecular signals underlying them, remained a much-coveted dream. Now, a new era of intravital fluorescence microscopy has dawned. In this Primer, we review the technologies that made this revolution possible, and demonstrate how intravital imaging is beginning to provide quantitative and dynamic insights into cell biology, immunology, tumor biolo...

  11. Intravital imaging.

    Science.gov (United States)

    Pittet, Mikael J; Weissleder, Ralph

    2011-11-23

    Until recently, the idea of observing life deep within the tissues of a living mouse, at a resolution sufficient to pick out cellular behaviors and molecular signals underlying them, remained a much-coveted dream. Now, a new era of intravital fluorescence microscopy has dawned. In this Primer, we review the technologies that made this revolution possible and demonstrate how intravital imaging is beginning to provide quantitative and dynamic insights into cell biology, immunology, tumor biology, and neurobiology. Copyright © 2011 Elsevier Inc. All rights reserved.

  12. Intravital multiphoton tomography as an appropriate tool for non-invasive in vivo analysis of human skin affected with atopic dermatitis

    Science.gov (United States)

    Huck, Volker; Gorzelanny, Christian; Thomas, Kai; Mess, Christian; Dimitrova, Valentina; Schwarz, Martin; Riemann, Iris; Niemeyer, Verena; Luger, Thomas A.; König, Karsten; Schneider, Stefan W.

    2011-03-01

    Increasing incidence of inflammatory skin diseases such as Atopic Dermatitis (AD) has been noted in the past years. According to recent estimations around 15% of newborn subjects are affected with a disease severity that requires medical treatment. Although its pathogenesis is multifactorial, recent reports indicate that an impaired physical skin barrier predispose for the development of AD. The major part of this barrier is formed by the stratum corneum (SC) wherein corneocytes are embedded in a complex matrix of proteins and lipids. Its components were synthesized in the stratum granulosum (SG) and secreted via lamellar bodies at the SC/SG interface. Within a clinical in vivo study we focused on the skin metabolism at the SC/SG interface in AD affected patients in comparison to healthy subjects. Measurement of fluorescence life-time of NADH provides access to the metabolic state of skin. Due to the application of a 5D intravital tomographic skin analysis we facilitate the non-invasive investigation of human epidermis in the longitudinal course of AD therapy. We could ascertain by blinded analysis of 40 skin areas of 20 patients in a three month follow-up that the metabolic status at the SC/SG interface was altered in AD compromised skin even in non-lesional, apparent healthy skin regions. This illustrates an impaired skin barrier formation even at non-affected skin of AD subjects appearing promotive for the development of acute skin inflammation. Therefore, our findings allow a deeper understanding of the individual disease development and the improved management of the therapeutic intervention in clinical application.

  13. Preparation of Murine Submandibular Salivary Gland for Upright Intravital Microscopy.

    Science.gov (United States)

    Ficht, Xenia; Thelen, Flavian; Stolp, Bettina; Stein, Jens V

    2018-05-07

    The submandibular salivary gland (SMG) is one of the three major salivary glands, and is of interest for many different fields of biological research, including cell biology, oncology, dentistry, and immunology. The SMG is an exocrine gland comprised of secretory epithelial cells, myofibroblasts, endothelial cells, nerves, and extracellular matrix. Dynamic cellular processes in the rat and mouse SMG have previously been imaged, mostly using inverted multi-photon microscope systems. Here, we describe a straightforward protocol for the surgical preparation and stabilization of the murine SMG in anesthetized mice for in vivo imaging with upright multi-photon microscope systems. We present representative intravital image sets of endogenous and adoptively transferred fluorescent cells, including the labeling of blood vessels or salivary ducts and second harmonic generation to visualize fibrillar collagen. In sum, our protocol allows for surgical preparation of mouse salivary glands in upright microscopy systems, which are commonly used for intravital imaging in the field of immunology.

  14. Video-rate resonant scanning multiphoton microscopy

    Science.gov (United States)

    Kirkpatrick, Nathaniel D.; Chung, Euiheon; Cook, Daniel C.; Han, Xiaoxing; Gruionu, Gabriel; Liao, Shan; Munn, Lance L.; Padera, Timothy P.; Fukumura, Dai; Jain, Rakesh K.

    2013-01-01

    The abnormal tumor microenvironment fuels tumor progression, metastasis, immune suppression, and treatment resistance. Over last several decades, developments in and applications of intravital microscopy have provided unprecedented insights into the dynamics of the tumor microenvironment. In particular, intravital multiphoton microscopy has revealed the abnormal structure and function of tumor-associated blood and lymphatic vessels, the role of aberrant tumor matrix in drug delivery, invasion and metastasis of tumor cells, the dynamics of immune cell trafficking to and within tumors, and gene expression in tumors. However, traditional multiphoton microscopy suffers from inherently slow imaging rates—only a few frames per second, thus unable to capture more rapid events such as blood flow, lymphatic flow, and cell movement within vessels. Here, we report the development and implementation of a video-rate multiphoton microscope (VR-MPLSM) based on resonant galvanometer mirror scanning that is capable of recording at 30 frames per second and acquiring intravital multispectral images. We show that the design of the system can be readily implemented and is adaptable to various experimental models. As examples, we demonstrate the utility of the system to directly measure flow within tumors, capture metastatic cancer cells moving within the brain vasculature and cells in lymphatic vessels, and image acute responses to changes in a vascular network. VR-MPLSM thus has the potential to further advance intravital imaging and provide new insight into the biology of the tumor microenvironment. PMID:24353926

  15. Multiphoton processes

    International Nuclear Information System (INIS)

    Manus, C.; Mainfray, G.

    1980-01-01

    The main features of multiphoton processes are described on a somewhat elementary basis. The emphasis is put on multiphoton ionization of atoms where the influence of resonance effects is given through typical examples. The important role played by the coherence of light is shown to produce a very dramatic influence on multiphoton absorption. Different observations concerning molecules, electrons, as well as solid surfaces illustrate the generality of these very non linear interaction between light and matter

  16. Multiphoton bibliography

    International Nuclear Information System (INIS)

    Eberly, J.H.; Gallagher, J.W.

    1981-12-01

    A bibliography is presented of approximately 275 references from literature published since 1980 on multiphoton research. A subject list is provided which divides the references into four subdivisions, i.e., ionization, bound-bound transitions, dissociation, and free-free transitions. An author index is included

  17. Multiphoton resonances

    International Nuclear Information System (INIS)

    Shore, B.W.

    1977-01-01

    The long-time average of level populations in a coherently-excited anharmonic sequence of energy levels (e.g., an anharmonic oscillator) exhibits sharp resonances as a function of laser frequency. For simple linearly-increasing anharmonicity, each resonance is a superposition of various multiphoton resonances (e.g., a superposition of 3, 5, 7, . . . photon resonances), each having its own characteristic width predictable from perturbation theory

  18. Multiphoton processes: conference proceedings

    International Nuclear Information System (INIS)

    Lambropoulos, P.; Smith, S.J.

    1984-01-01

    The chapters of this volume represent the invited papers delivered at the conference. They are arranged according to thermatic proximity beginning with atoms and continuing with molecules and surfaces. Section headings include multiphoton processes in atoms, field fluctuations and collisions in multiphoton process, and multiphoton processes in molecules and surfaces. Abstracts of individual items from the conference were prepared separately for the data base

  19. Multiphoton processes: conference proceedings

    Energy Technology Data Exchange (ETDEWEB)

    Lambropoulos, P.; Smith, S.J. (eds.)

    1984-01-01

    The chapters of this volume represent the invited papers delivered at the conference. They are arranged according to thermatic proximity beginning with atoms and continuing with molecules and surfaces. Section headings include multiphoton processes in atoms, field fluctuations and collisions in multiphoton process, and multiphoton processes in molecules and surfaces. Abstracts of individual items from the conference were prepared separately for the data base. (GHT)

  20. High-resolution intravital microscopy.

    Directory of Open Access Journals (Sweden)

    Volker Andresen

    Full Text Available Cellular communication constitutes a fundamental mechanism of life, for instance by permitting transfer of information through synapses in the nervous system and by leading to activation of cells during the course of immune responses. Monitoring cell-cell interactions within living adult organisms is crucial in order to draw conclusions on their behavior with respect to the fate of cells, tissues and organs. Until now, there is no technology available that enables dynamic imaging deep within the tissue of living adult organisms at sub-cellular resolution, i.e. detection at the level of few protein molecules. Here we present a novel approach called multi-beam striped-illumination which applies for the first time the principle and advantages of structured-illumination, spatial modulation of the excitation pattern, to laser-scanning-microscopy. We use this approach in two-photon-microscopy--the most adequate optical deep-tissue imaging-technique. As compared to standard two-photon-microscopy, it achieves significant contrast enhancement and up to 3-fold improved axial resolution (optical sectioning while photobleaching, photodamage and acquisition speed are similar. Its imaging depth is comparable to multifocal two-photon-microscopy and only slightly less than in standard single-beam two-photon-microscopy. Precisely, our studies within mouse lymph nodes demonstrated 216% improved axial and 23% improved lateral resolutions at a depth of 80 µm below the surface. Thus, we are for the first time able to visualize the dynamic interactions between B cells and immune complex deposits on follicular dendritic cells within germinal centers (GCs of live mice. These interactions play a decisive role in the process of clonal selection, leading to affinity maturation of the humoral immune response. This novel high-resolution intravital microscopy method has a huge potential for numerous applications in neurosciences, immunology, cancer research and

  1. High-Resolution Intravital Microscopy

    Science.gov (United States)

    Andresen, Volker; Pollok, Karolin; Rinnenthal, Jan-Leo; Oehme, Laura; Günther, Robert; Spiecker, Heinrich; Radbruch, Helena; Gerhard, Jenny; Sporbert, Anje; Cseresnyes, Zoltan; Hauser, Anja E.; Niesner, Raluca

    2012-01-01

    Cellular communication constitutes a fundamental mechanism of life, for instance by permitting transfer of information through synapses in the nervous system and by leading to activation of cells during the course of immune responses. Monitoring cell-cell interactions within living adult organisms is crucial in order to draw conclusions on their behavior with respect to the fate of cells, tissues and organs. Until now, there is no technology available that enables dynamic imaging deep within the tissue of living adult organisms at sub-cellular resolution, i.e. detection at the level of few protein molecules. Here we present a novel approach called multi-beam striped-illumination which applies for the first time the principle and advantages of structured-illumination, spatial modulation of the excitation pattern, to laser-scanning-microscopy. We use this approach in two-photon-microscopy - the most adequate optical deep-tissue imaging-technique. As compared to standard two-photon-microscopy, it achieves significant contrast enhancement and up to 3-fold improved axial resolution (optical sectioning) while photobleaching, photodamage and acquisition speed are similar. Its imaging depth is comparable to multifocal two-photon-microscopy and only slightly less than in standard single-beam two-photon-microscopy. Precisely, our studies within mouse lymph nodes demonstrated 216% improved axial and 23% improved lateral resolutions at a depth of 80 µm below the surface. Thus, we are for the first time able to visualize the dynamic interactions between B cells and immune complex deposits on follicular dendritic cells within germinal centers (GCs) of live mice. These interactions play a decisive role in the process of clonal selection, leading to affinity maturation of the humoral immune response. This novel high-resolution intravital microscopy method has a huge potential for numerous applications in neurosciences, immunology, cancer research and developmental biology

  2. Intravital live cell triggered imaging system reveals monocyte patrolling and macrophage migration in atherosclerotic arteries

    Science.gov (United States)

    McArdle, Sara; Chodaczek, Grzegorz; Ray, Nilanjan; Ley, Klaus

    2015-02-01

    Intravital multiphoton imaging of arteries is technically challenging because the artery expands with every heartbeat, causing severe motion artifacts. To study leukocyte activity in atherosclerosis, we developed the intravital live cell triggered imaging system (ILTIS). This system implements cardiac triggered acquisition as well as frame selection and image registration algorithms to produce stable movies of myeloid cell movement in atherosclerotic arteries in live mice. To minimize tissue damage, no mechanical stabilization is used and the artery is allowed to expand freely. ILTIS performs multicolor high frame-rate two-dimensional imaging and full-thickness three-dimensional imaging of beating arteries in live mice. The external carotid artery and its branches (superior thyroid and ascending pharyngeal arteries) were developed as a surgically accessible and reliable model of atherosclerosis. We use ILTIS to demonstrate Cx3cr1GFP monocytes patrolling the lumen of atherosclerotic arteries. Additionally, we developed a new reporter mouse (Apoe-/-Cx3cr1GFP/+Cd11cYFP) to image GFP+ and GFP+YFP+ macrophages "dancing on the spot" and YFP+ macrophages migrating within intimal plaque. ILTIS will be helpful to answer pertinent open questions in the field, including monocyte recruitment and transmigration, macrophage and dendritic cell activity, and motion of other immune cells.

  3. Correlative intravital imaging of cGMP signals and vasodilation in mice

    Directory of Open Access Journals (Sweden)

    Martin eThunemann

    2014-10-01

    Full Text Available Cyclic guanosine monophosphate (cGMP is an important signaling molecule and drug target in the cardiovascular system. It is well known that stimulation of the vascular nitric oxide (NO-cGMP pathway results in vasodilation. However, the spatiotemporal dynamics of cGMP signals themselves and the cGMP concentrations within specific cardiovascular cell types in health, disease, and during pharmacotherapy with cGMP-elevating drugs are largely unknown. To facilitate the analysis of cGMP signaling in vivo, we have generated transgenic mice that express fluorescence resonance energy transfer (FRET-based cGMP sensor proteins. Here, we describe two models of intravital FRET/cGMP imaging in the vasculature of cGMP sensor mice: (1 epifluorescence-based ratio imaging in resistance-type vessels of the cremaster muscle and (2 ratio imaging by multiphoton microscopy within the walls of subcutaneous blood vessels accessed through a dorsal skinfold chamber. Both methods allow simultaneous monitoring of NO-induced cGMP transients and vasodilation in living mice. Detailed protocols of all steps necessary to perform and evaluate intravital imaging experiments of the vasculature of anesthetized mice including surgery, imaging, and data evaluation are provided. An image segmentation approach is described to estimate FRET/cGMP changes within moving structures such as the vessel wall during vasodilation. The methods presented herein should be useful to visualize cGMP or other biochemical signals that are detectable with FRET-based biosensors, such as cyclic adenosine monophosphate or Ca2+, and to correlate them with respective vascular responses. With further refinement and combination of transgenic mouse models and intravital imaging technologies, we envision an exciting future, in which we are able to ‘watch’ biochemistry, (patho physiology, and pharmacotherapy in the context of a living mammalian organism.

  4. Fluorescein Derivatives in Intravital Fluorescence Imaging

    Directory of Open Access Journals (Sweden)

    Michael S. Roberts

    2013-08-01

    Full Text Available Intravital fluorescence microscopy enables the direct imaging of fluorophores in vivo and advanced techniques such as fluorescence lifetime imaging (FLIM enable the simultaneous detection of multiple fluorophores. Consequently, it is now possible to record distribution and metabolism of a chemical in vivo and to optimise the delivery of fluorophores in vivo. Recent clinical applications with fluorescein and other intravital fluorescent stains have occurred in neurosurgery, dermatology [including photodynamic therapy (PDT] and endomicroscopy. Potential uses have been identified in periodontal disease, skin graft and cancer surgery. Animal studies have demonstrated that diseased tissue can be specifically stained with fluorophore conjugates. This review focuses on the fluorescein derived fluorophores in common clinical use and provides examples of novel applications from studies in tissue samples.

  5. In vivo multiphoton imaging of bile duct ligation

    Science.gov (United States)

    Liu, Yuan; Li, Feng-Chieh; Chen, Hsiao-Chin; Chang, Po-shou; Yang, Shu-Mei; Lee, Hsuan-Shu; Dong, Chen-Yuan

    2008-02-01

    Bile is the exocrine secretion of liver and synthesized by hepatocytes. It is drained into duodenum for the function of digestion or drained into gallbladder for of storage. Bile duct obstruction is a blockage in the tubes that carry bile to the gallbladder and small intestine. However, Bile duct ligation results in the changes of bile acids in serum, liver, urine, and feces1, 2. In this work, we demonstrate a novel technique to image this pathological condition by using a newly developed in vivo imaging system, which includes multiphoton microscopy and intravital hepatic imaging chamber. The images we acquired demonstrate the uptake, processing of 6-CFDA in hepatocytes and excretion of CF in the bile canaliculi. In addition to imaging, we can also measure kinetics of the green fluorescence intensity.

  6. Multi-Photon Entanglement and Quantum Teleportation

    National Research Council Canada - National Science Library

    Shih, Yanhua

    1999-01-01

    The project 'Multi-Photon Entanglement and Quantum Teleportation' concerns a series of experimental and theoretical investigations on multi-photon entangled states and the applications, for example...

  7. Clinical multiphoton FLIM tomography

    Science.gov (United States)

    König, Karsten

    2012-03-01

    This paper gives an overview on current clinical high resolution multiphoton fluorescence lifetime imaging in volunteers and patients. Fluorescence lifetime imaging (FLIM) in Life Sciences was introduced in Jena/Germany in 1988/89 based on a ZEISS confocal picosecond dye laser scanning microscope equipped with a single photon counting unit. The porphyrin distribution in living cells and living tumor-bearing mice was studied with high spatial, temporal, and spectral resolution. Ten years later, time-gated cameras were employed to detect dental caries in volunteers based on one-photon excitation of autofluorescent bacteria with long fluorescence lifetimes. Nowadays, one-photon FLIM based on picosecond VIS laser diodes are used to study ocular diseases in humans. Already one decade ago, first clinical twophoton FLIM images in humans were taken with the certified clinical multiphoton femtosecond laser tomograph DermaInspectTM. Multiphoton tomographs with FLIM modules are now operating in hospitals at Brisbane, Tokyo, Berlin, Paris, London, Modena and other European cities. Multiple FLIM detectors allow spectral FLIM with a temporal resolution down to 20 ps (MCP) / 250 ps (PMT) and a spectral resolution of 10 nm. Major FLIM applications include the detection of intradermal sunscreen and tattoo nanoparticles, the detection of different melanin types, the early diagnosis of dermatitis and malignant melanoma, as well as the measurement of therapeutic effects in pateints suffering from dermatitis. So far, more than 1,000 patients and volunteers have been investigated with the clinical multiphoton FLIM tomographs DermaInspectTM and MPTflexTM.

  8. Generalized Multiphoton Quantum Interference

    Directory of Open Access Journals (Sweden)

    Max Tillmann

    2015-10-01

    Full Text Available Nonclassical interference of photons lies at the heart of optical quantum information processing. Here, we exploit tunable distinguishability to reveal the full spectrum of multiphoton nonclassical interference. We investigate this in theory and experiment by controlling the delay times of three photons injected into an integrated interferometric network. We derive the entire coincidence landscape and identify transition matrix immanants as ideally suited functions to describe the generalized case of input photons with arbitrary distinguishability. We introduce a compact description by utilizing a natural basis that decouples the input state from the interferometric network, thereby providing a useful tool for even larger photon numbers.

  9. Doubly resonant multiphoton ionization

    International Nuclear Information System (INIS)

    Crance, M.

    1978-01-01

    A particular case of doubly resonant multiphoton ionization is theoretically investigated. More precisely, two levels quasi-resonant with two successive harmonics of the field frequency are considered. The method used is based on the effective operator formalism first introduced for this problem by Armstrong, Beers and Feneuille. The main result is to show the possibility of observing large interference effects on the width of the resonances. Moreover this treatment allows us to make more precise the connection between effective operator formalism and standard perturbation theory

  10. Intravital Microscopic Interrogation of Peripheral Taste Sensation

    Science.gov (United States)

    Choi, Myunghwan; Lee, Woei Ming; Yun, Seok Hyun

    2015-03-01

    Intravital microscopy is a powerful tool in neuroscience but has not been adapted to the taste sensory organ due to anatomical constraint. Here we developed an imaging window to facilitate microscopic access to the murine tongue in vivo. Real-time two-photon microscopy allowed the visualization of three-dimensional microanatomy of the intact tongue mucosa and functional activity of taste cells in response to topically administered tastants in live mice. Video microscopy also showed the calcium activity of taste cells elicited by small-sized tastants in the blood circulation. Molecular kinetic analysis suggested that intravascular taste sensation takes place at the microvilli on the apical side of taste cells after diffusion of the molecules through the pericellular capillaries and tight junctions in the taste bud. Our results demonstrate the capabilities and utilities of the new tool for taste research in vivo.

  11. Transverse correlations in multiphoton entanglement

    International Nuclear Information System (INIS)

    Wen Jianming; Rubin, Morton H.; Shih Yanhua

    2007-01-01

    We have analyzed the transverse correlation in multiphoton entanglement. The generalization of quantum ghost imaging is extended to the N-photon state. The Klyshko's two-photon advanced-wave picture is generalized to the N-photon case

  12. Multiphoton Processes and Attosecond Physics

    CERN Document Server

    Midorikawa, Katsumi; 12th International Conference on Multiphoton Processes; 3rd International Conference on Attosecond Physics

    2012-01-01

    Recent advances in ultrashort pulsed laser technology have opened new frontiers in atomic, molecular and optical sciences. The 12th International Conference on Multiphoton Processes (ICOMP12) and the 3rd International Conference on Attosecond Physics (ATTO3), held jointly in Sapporo, Japan, during July 3-8, showcased studies at the forefront of research on multiphoton processes and attosecond physics. This book summarizes presentations and discussions from these two conferences.

  13. Laser-induced multiphoton transitions

    International Nuclear Information System (INIS)

    Stenholm, S.

    1978-06-01

    Laser induced multiphoton processes are reviewed. The effects of strong fields on atoms are discussed. The perturbation treatment is presented and also its generalization to treat intermediate resonances. The influence of atomic coherence is discussed heuristically and the relation between quantal and classical descriptions of the field is elucidated by reference to the dressed atom description. Atomic ionization experiments are reviewed and the present understanding of multiphoton dissociation of molecules is explained. Finally some prospects for the future are discussed. (author)

  14. Six-color intravital two-photon imaging of brain tumors and their dynamic microenvironment

    Directory of Open Access Journals (Sweden)

    Clément eRicard

    2014-02-01

    Full Text Available The majority of intravital studies on brain tumor in living animal so far rely on dual color imaging. We describe here a multiphoton imaging protocol to dynamically characterize the interactions between six cellular components in a living mouse. We applied this methodology to a clinically relevant glioblastoma multiforme (GBM model designed in reporter mice with targeted cell populations labeled by fluorescent proteins of different colors. This model permitted us to make non-invasive longitudinal and multi-scale observations of cell-to-cell interactions. We provide examples of such 5D (x,y,z,t,color images acquired on a daily basis from volumes of interest, covering most of the mouse parietal cortex at subcellular resolution. Spectral deconvolution allowed us to accurately separate of each cell population as well as some components of the extracellular matrix. The technique represents a powerful tool for investigating how tumor progression is influenced by the interactions of tumor cells with host cells and the extracellular matrix micro-environment. It will be especially valuable for evaluating neuro-oncological drug efficacy and target specificity. The imaging protocol provided here can be easily translated to other mouse models of neuropathologies, and should also be of fundamental interest for investigations in other areas of systems biology.

  15. Six-color intravital two-photon imaging of brain tumors and their dynamic microenvironment.

    Science.gov (United States)

    Ricard, Clément; Debarbieux, Franck Christian

    2014-01-01

    The majority of intravital studies on brain tumor in living animal so far rely on dual color imaging. We describe here a multiphoton imaging protocol to dynamically characterize the interactions between six cellular components in a living mouse. We applied this methodology to a clinically relevant glioblastoma multiforme (GBM) model designed in reporter mice with targeted cell populations labeled by fluorescent proteins of different colors. This model permitted us to make non-invasive longitudinal and multi-scale observations of cell-to-cell interactions. We provide examples of such 5D (x,y,z,t,color) images acquired on a daily basis from volumes of interest, covering most of the mouse parietal cortex at subcellular resolution. Spectral deconvolution allowed us to accurately separate each cell population as well as some components of the extracellular matrix. The technique represents a powerful tool for investigating how tumor progression is influenced by the interactions of tumor cells with host cells and the extracellular matrix micro-environment. It will be especially valuable for evaluating neuro-oncological drug efficacy and target specificity. The imaging protocol provided here can be easily translated to other mouse models of neuropathologies, and should also be of fundamental interest for investigations in other areas of systems biology.

  16. Quantitative multiphoton imaging

    Science.gov (United States)

    König, Karsten; Weinigel, Martin; Breunig, Hans Georg; Uchugonova, Aisada

    2014-02-01

    Certified clinical multiphoton tomographs for label-free multidimensional high-resolution in vivo imaging have been introduced to the market several years ago. Novel tomographs include a flexible 360° scan head attached to a mechanooptical arm for autofluorescence and SHG imaging as well as a CARS module. Non-fluorescent lipids and water, mitochondrial fluorescent NAD(P)H, fluorescent elastin, keratin, and melanin as well as SHG-active collagen can be imaged in vivo with submicron resolution in human skin. Sensitive and rapid detectors allow single photon counting and the construction of 3D maps where the number of detected photons per voxel is depicted. Intratissue concentration profiles from endogenous as well exogenous substances can be generated when the number of detected photons can be correlated with the number of molecules with respect to binding and scattering behavior. Furthermore, the skin ageing index SAAID based on the ratio elastin/collagen as well as the epidermis depth based on the onset of SHG generation can be determined.

  17. 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.

  18. Using multiphoton fluorescence lifetime imaging to characterize liver damage and fluorescein disposition in liver in vivo

    Science.gov (United States)

    Thorling, Camilla A.; Studier, Hauke; Crawford, Darrell; Roberts, Michael S.

    2016-03-01

    Liver disease is the fifth most common cause of death and unlike many other major causes of mortality, liver disease rates are increasing rather than decreasing. There is no ideal measurement of liver disease and although biopsies are the gold standard, this only allows for a spot examination and cannot follow dynamic processes of the liver. Intravital imaging has the potential to extract detailed information over a larger sampling area continuously. The aim of this project was to investigate whether multiphoton and fluorescence lifetime imaging microscopy could detect early liver damage and to assess whether it could detect changes in metabolism of fluorescein in normal and diseased livers. Four experimental groups were used in this study: 1) control; 2) ischemia reperfusion injury; 3) steatosis and 4) steatosis with ischemia reperfusion injury. Results showed that multiphoton microscopy could visualize morphological changes such as decreased fluorescence of endogenous fluorophores and the presence of lipid droplets, characteristic of steatosis. Fluorescence lifetime imaging microscopy showed increase in NADPH in steatosis with and without ischemia reperfusion injury and could detect changes in metabolism of fluorescein to fluorescein monoglurcuronide, which was impaired in steatosis with ischemia reperfusion injury. These results concluded that the combination of multiphoton microscopy and fluorescence lifetime imaging is a promising method of assessing early stage liver damage and that it can be used to study changes in drug metabolism in the liver as an indication of liver disease and has the potential to replace the traditional static liver biopsy currently used.

  19. Microhemodynamic parameters quantification from intravital microscopy videos

    International Nuclear Information System (INIS)

    Ortiz, Daniel; Cabrales, Pedro; Briceño, Juan Carlos

    2014-01-01

    Blood flow and blood–endothelium interactions correspond with the genesis of cardiovascular diseases. Therefore, quantitative analysis of blood flow dynamics at the microcirculation level is of special interest. Regulatory mechanisms mediated by blow flow have been studied in detail using in vitro approaches. However, these mechanisms have not been fully validated in vivo due to technical limitations that arise when quantifying microhemodynamics with the required level of detail. Intravital microscopy combined with high-speed video recordings has been used for the analysis of blood flow in small blood vessels of chronic and acute experimental tissue preparations. This tool can be used to study the interaction between the flowing blood and the vessel walls of arterioles and venules with sufficient temporal and spatial resolution. Our objective was to develop a simple and robust cross-correlation algorithm for the automatic analysis of high-speed video recordings of microcirculatory blood flow. The algorithm was validated using in vitro and in vivo systems. Results indicate that the algorithm's ability to estimate the velocity of local red blood cells as a function of blood vessel radius is highly accurate. They thereby suggest that the algorithm could be used to explore dynamic changes in blood flow under different experimental conditions including a wide range of flow rates and hematocrit levels. The algorithm can also be used to measure volumetric flow rates, radial velocity profiles, wall shear rate, and wall shear stress. Several applications are presently explored, including the analysis of velocity profiles in the branches of arterial bifurcations. This work demonstrates the robustness of the cross-correlation technique in various flow conditions and elucidates its potential application for in vivo determination of blood flow dynamics in the microcirculation. (paper)

  20. Multiphoton microscopy imaging of developing tooth germs

    Directory of Open Access Journals (Sweden)

    Pei-Yu Pan

    2014-01-01

    Conclusion: In this study, a novel multiphoton microscopy database of images from developing tooth germs in mice was set up. We confirmed that multiphoton laser microscopy is a powerful tool for investigating the development of tooth germ and is worthy for further application in the study of tooth regeneration.

  1. Signal improvement in multiphoton microscopy by reflection with simple mirrors near the sample

    Science.gov (United States)

    Rehberg, Markus; Krombach, Fritz; Pohl, Ulrich; Dietzel, Steffen

    2010-03-01

    In conventional fluorescence or confocal microscopy, emitted light is generated not only in the focal plane but also above and below. The situation is different in multiphoton-induced fluorescence and multiphoton-induced higher harmonic generation. Here, restriction of signal generation to a single focal point permits that all emitted photons can contribute to image formation if collected, regardless of their path through the specimen. Often, the intensity of the emitted light is rather low in biological specimens. We present a method to significantly increase the fraction of photons collected by an epi (backward) detector by placing a simple mirror, an aluminum-coated coverslip, directly under the sample. Samples investigated include fluorescent test slides, collagen gels, and thin-layered, intact mouse skeletal muscles. Quantitative analysis revealed an intensity increase of second- and third-harmonic generated signal in skeletal muscle of nine- and sevenfold respectively, and of fluorescent signal in test slides of up to twofold. Our approach thus allows significant signal improvement also for situations were a forward detection is impossible, e.g., due to the anatomy of animals in intravital microscopy.

  2. Multiphoton quantum optics and quantum state engineering

    Energy Technology Data Exchange (ETDEWEB)

    Dell' Anno, Fabio [Dipartimento di Fisica ' E. R. Caianiello' , Universita degli Studi di Salerno, CNISM and CNR-INFM Coherentia, and INFN Sezione di Napoli, Gruppo Collegato di Salerno, Via S. Allende, I-84081 Baronissi (Saudi Arabia) (Italy)]. E-mail: dellanno@sa.infn.it; De Siena, Silvio [Dipartimento di Fisica ' E. R. Caianiello' , Universita degli Studi di Salerno, CNISM and CNR-INFM Coherentia, and INFN Sezione di Napoli, Gruppo Collegato di Salerno, Via S. Allende, I-84081 Baronissi (SA) (Italy)]. E-mail: desiena@sa.infn.it; Illuminati, Fabrizio [Dipartimento di Fisica ' E. R. Caianiello' , Universita degli Studi di Salerno, CNISM and CNR-INFM Coherentia, and INFN Sezione di Napoli, Gruppo Collegato di Salerno, Via S. Allende, I-84081 Baronissi (SA) (Italy)]. E-mail: illuminati@sa.infn.it

    2006-05-15

    We present a review of theoretical and experimental aspects of multiphoton quantum optics. Multiphoton processes occur and are important for many aspects of matter-radiation interactions that include the efficient ionization of atoms and molecules, and, more generally, atomic transition mechanisms; system-environment couplings and dissipative quantum dynamics; laser physics, optical parametric processes, and interferometry. A single review cannot account for all aspects of such an enormously vast subject. Here we choose to concentrate our attention on parametric processes in nonlinear media, with special emphasis on the engineering of nonclassical states of photons and atoms that are relevant for the conceptual investigations as well as for the practical applications of forefront aspects of modern quantum mechanics. We present a detailed analysis of the methods and techniques for the production of genuinely quantum multiphoton processes in nonlinear media, and the corresponding models of multiphoton effective interactions. We review existing proposals for the classification, engineering, and manipulation of nonclassical states, including Fock states, macroscopic superposition states, and multiphoton generalized coherent states. We introduce and discuss the structure of canonical multiphoton quantum optics and the associated one- and two-mode canonical multiphoton squeezed states. This framework provides a consistent multiphoton generalization of two-photon quantum optics and a consistent Hamiltonian description of multiphoton processes associated to higher-order nonlinearities. Finally, we discuss very recent advances that by combining linear and nonlinear optical devices allow to realize multiphoton entangled states of the electromagnetic field, either in discrete or in continuous variables, that are relevant for applications to efficient quantum computation, quantum teleportation, and related problems in quantum communication and information.

  3. Multiphoton quantum optics and quantum state engineering

    International Nuclear Information System (INIS)

    Dell'Anno, Fabio; De Siena, Silvio; Illuminati, Fabrizio

    2006-01-01

    We present a review of theoretical and experimental aspects of multiphoton quantum optics. Multiphoton processes occur and are important for many aspects of matter-radiation interactions that include the efficient ionization of atoms and molecules, and, more generally, atomic transition mechanisms; system-environment couplings and dissipative quantum dynamics; laser physics, optical parametric processes, and interferometry. A single review cannot account for all aspects of such an enormously vast subject. Here we choose to concentrate our attention on parametric processes in nonlinear media, with special emphasis on the engineering of nonclassical states of photons and atoms that are relevant for the conceptual investigations as well as for the practical applications of forefront aspects of modern quantum mechanics. We present a detailed analysis of the methods and techniques for the production of genuinely quantum multiphoton processes in nonlinear media, and the corresponding models of multiphoton effective interactions. We review existing proposals for the classification, engineering, and manipulation of nonclassical states, including Fock states, macroscopic superposition states, and multiphoton generalized coherent states. We introduce and discuss the structure of canonical multiphoton quantum optics and the associated one- and two-mode canonical multiphoton squeezed states. This framework provides a consistent multiphoton generalization of two-photon quantum optics and a consistent Hamiltonian description of multiphoton processes associated to higher-order nonlinearities. Finally, we discuss very recent advances that by combining linear and nonlinear optical devices allow to realize multiphoton entangled states of the electromagnetic field, either in discrete or in continuous variables, that are relevant for applications to efficient quantum computation, quantum teleportation, and related problems in quantum communication and information

  4. Multiphoton Microscopy for Ophthalmic Imaging

    Directory of Open Access Journals (Sweden)

    Emily A. Gibson

    2011-01-01

    Full Text Available We review multiphoton microscopy (MPM including two-photon autofluorescence (2PAF, second harmonic generation (SHG, third harmonic generation (THG, fluorescence lifetime (FLIM, and coherent anti-Stokes Raman Scattering (CARS with relevance to clinical applications in ophthalmology. The different imaging modalities are discussed highlighting the particular strength that each has for functional tissue imaging. MPM is compared with current clinical ophthalmological imaging techniques such as reflectance confocal microscopy, optical coherence tomography, and fluorescence imaging. In addition, we discuss the future prospects for MPM in disease detection and clinical monitoring of disease progression, understanding fundamental disease mechanisms, and real-time monitoring of drug delivery.

  5. Tracking the stochastic fate of cells of the renin lineage after podocyte depletion using multicolor reporters and intravital imaging.

    Directory of Open Access Journals (Sweden)

    Natalya V Kaverina

    Full Text Available Podocyte depletion plays a major role in focal segmental glomerular sclerosis (FSGS. Because cells of the renin lineage (CoRL serve as adult podocyte and parietal epithelial cell (PEC progenitor candidates, we generated Ren1cCre/R26R-ConfettiTG/WT and Ren1dCre/R26R-ConfettiTG/WT mice to determine CoRL clonality during podocyte replacement. Four CoRL reporters (GFP, YFP, RFP, CFP were restricted to cells in the juxtaglomerular compartment (JGC at baseline. Following abrupt podocyte depletion in experimental FSGS, all four CoRL reporters were detected in a subset of glomeruli at day 28, where they co-expressed de novo four podocyte proteins (podocin, nephrin, WT-1 and p57 and two glomerular parietal epithelial cell (PEC proteins (claudin-1, PAX8. To monitor the precise migration of a subset of CoRL over a 2w period following podocyte depletion, intravital multiphoton microscopy was used. Our findings demonstrate direct visual support for the migration of single CoRL from the JGC to the parietal Bowman's capsule, early proximal tubule, mesangium and glomerular tuft. In summary, these results suggest that following podocyte depletion, multi-clonal CoRL migrate to the glomerulus and replace podocyte and PECs in experimental FSGS.

  6. Multiphoton dissociation of polyatomic molecules

    International Nuclear Information System (INIS)

    Schulz, P.A.

    1979-10-01

    The dynamics of infrared multiphoton excitation and dissociation of SF 6 was investigated under collision free conditions by a crossed laser-molecular beam method. In order to understand the excitation mechanism and to elucidate the requirements of laser intensity and energy fluence, a series of experiments were carried out to measure the dissociation yield dependences on energy fluence, vibrational temperature of SF 6 , the pulse duration of the CO 2 laser and the frequency in both one and two laser experiments. Translational energy distributions of the SF 5 dissociation product measured by time of flight and angular distributions and the dissociation lifetime of excited SF 6 as inferred from the observation of secondary dissociation of SF 5 into SF 4 and F during the laser pulse suggest that the dynamics of dissociation of excited molecules is dominated by complete energy randomization and rapid intramolecular energy transfer on a nanosecond timescale, and can be adequately described by RRKM theory. An improved phenomenological model including the initial intensity dependent excitation, a rate equation describing the absorption and stimulated emission of single photons, and the unimolecular dissociation of excited molecules is constructed based on available experimental results. The model shows that the energy fluence of the laser determines the excitation of molecules in the quasi-continuum and the excess energy with which molecules dissociate after the laser pulse. The role played by the laser intensity in multiphoton dissociation is more significant than just that of overcoming the intensity dependent absorption in the lowest levels. 63 references

  7. MULTI-PHOTON PHOSPHOR FEASIBILITY RESEARCH

    Energy Technology Data Exchange (ETDEWEB)

    R. Graham; W. Chow

    2003-05-01

    Development of multi-photon phosphor materials for discharge lamps represents a goal that would achieve up to a doubling of discharge (fluorescent) lamp efficacy. This report reviews the existing literature on multi-photon phosphors, identifies obstacles in developing such phosphors, and recommends directions for future research to address these obstacles. To critically examine issues involved in developing a multi-photon phosphor, the project brought together a team of experts from universities, national laboratories, and an industrial lamp manufacturer. Results and findings are organized into three categories: (1) Multi-Photon Systems and Processes, (2) Chemistry and Materials Issues, and (3) Concepts and Models. Multi-Photon Systems and Processes: This category focuses on how to use our current understanding of multi-photon phosphor systems to design new phosphor systems for application in fluorescent lamps. The quickest way to develop multi-photon lamp phosphors lies in finding sensitizer ions for Gd{sup 3+} and identifying activator ions to red shift the blue emission from Pr{sup 3+} due to the {sup 1}S{sub 0} {yields} {sup 1}I{sub 6} transition associated with the first cascading step. Success in either of these developments would lead to more efficient fluorescent lamps. Chemistry and Materials Issues: The most promising multi-photon phosphors are found in fluoride hosts. However, stability of fluorides in environments typically found in fluorescent lamps needs to be greatly improved. Experimental investigation of fluorides in actual lamp environments needs to be undertaken while working on oxide and oxyfluoride alternative systems for backup. Concepts and Models: Successful design of a multi-photon phosphor system based on cascading transitions of Gd{sup 3+} and Pr{sup 3+} depends critically on how the former can be sensitized and the latter can sensitize an activator ion. Methods to predict energy level diagrams and Judd-Ofelt parameters of multi-photon

  8. Morphological signs of the intravital contraction (retraction of thrombotic emboli

    Directory of Open Access Journals (Sweden)

    R R Khismatullin

    2018-02-01

    Full Text Available Aim. To establish whether contraction (retraction of thrombi and thrombotic emboli occurs in vivo using structural signs of blood clot compression, such as compressive deformation of erythrocytes and redistribution of fibrin on the surface of a clot. Methods. Three postmortem pulmonary thrombotic emboli were examined by scanning electron microscopy and light microscopy after staining with hematoxylin and eosin as well as with Mallory’s method. Results. In 2 studied pulmonary emboli, extracted 7 and 15 hours after patients’ death, polyhedral erythrocytes (polyhedrocytes were revealed that were formed as a result of mechanical deformation under the action of contractile forces generated by activated platelets. In addition, the uneven distribution of fibrin within the emboli was found with displacement of fibrin to the periphery of the emboli, which is characteristic for contracted blood clot. In the first and the «oldest» clot extracted 38 hours after the patient’s death, the described contraction signs were absent, which was likely related to the postmortem autolysis or intravital pathological impairment of contraction. Conclusion. Thrombotic emboli ex vivo have morphological signs of contraction, suggesting intravital compression of the primary thrombi and/or thrombotic emboli, which might be an important pathogenetic mechanism for modulation of impaired blood flow at the sites of thrombotic occlusion of a vessel; the presence or absence of compressed erythrocytes inside and predominant location of fibrin on the periphery of a thrombus or embolus can potentially serve as additional pathomorphological criteria of death coming prescription.

  9. Imaging windows for long-term intravital imaging

    Science.gov (United States)

    Alieva, Maria; Ritsma, Laila; Giedt, Randy J; Weissleder, Ralph; van Rheenen, Jacco

    2014-01-01

    Intravital microscopy is increasingly used to visualize and quantitate dynamic biological processes at the (sub)cellular level in live animals. By visualizing tissues through imaging windows, individual cells (e.g., cancer, host, or stem cells) can be tracked and studied over a time-span of days to months. Several imaging windows have been developed to access tissues including the brain, superficial fascia, mammary glands, liver, kidney, pancreas, and small intestine among others. Here, we review the development of imaging windows and compare the most commonly used long-term imaging windows for cancer biology: the cranial imaging window, the dorsal skin fold chamber, the mammary imaging window, and the abdominal imaging window. Moreover, we provide technical details, considerations, and trouble-shooting tips on the surgical procedures and microscopy setups for each imaging window and explain different strategies to assure imaging of the same area over multiple imaging sessions. This review aims to be a useful resource for establishing the long-term intravital imaging procedure. PMID:28243510

  10. Optical Magnetometry Using Multiphoton Transitions

    Science.gov (United States)

    Degenkolb, Skyler M.

    Optical magnetometry plays a critical role in low-energy precision measurements and numerous other applications. In particular, permanent electric dipole moment (EDM) searches impose strict requirements on magnetic field sensitivity of the underlying atomic or molecular species. Other magnetometer properties - such as chemical reactivity, dielectric strength, and interaction cross-sections with other species - also impose limitations on experimental conditions. Here, we explore a novel approach to optical magnetometry, using multiphoton transitions of diamagnetic atoms to detect Larmor precession of polarized nuclei. Resonant probes are possible at moderate ultraviolet wavelengths, and hyperfine structure couples spin precession to fluorescence transitions with negligible backgrounds; paramagnetic rotation due to intensity-dependent dispersion may also be detectable. Nuclear spins and nonlinear optical excitation introduce new degrees of freedom, and evade limitations arising from rapid electronic decoherence. This dissertation reports progress towards two-photon optical magnetometry using ytterbium, rubidium, and xenon. We characterize the influence of probe polarization and magnetic fields on fluorescence spectra, for one- and two-photon continuous-wave (cw) excitation of ytterbium. Resolved hyperfine and isotope structure allow us to use spin-zero isotopes for diagnostics and normalization, and we develop analysis for overlapping two-photon resonances. We also report measurements of two-photon excitation in ytterbium and rubidium using picosecond laser pulses, and in xenon using a cw laser. Although hyperfine structure is unresolved, the rubidium measurements are sensitive to probe field polarization. Fluorescence spectra from two-photon excitation of ytterbium with femtosecond pulses show modulation when the repetition rate changes. Although techniques for polarizing noble gas nuclei are mature, existing cell designs are incompatible with two

  11. Current developments in clinical multiphoton tomography

    Science.gov (United States)

    König, Karsten; Weinigel, Martin; Breunig, Hans Georg; Gregory, Axel; Fischer, Peter; Kellner-Höfer, Marcel; Bückle, Rainer

    2010-02-01

    Two-photon microscopy has been introduced in 1990 [1]. 13 years later, CE-marked clinical multiphoton systems for 3D imaging of human skin with subcellular resolution have been launched by the JenLab company with the tomograph DermaInspectTM. In 2010, the second generation of clinical multiphoton tomographs was introduced. The novel mobile multiphoton tomograph MPTflexTM, equipped with a flexible articulated optical arm, provides an increased flexibility and accessibility especially for clinical and cosmetical examinations. The multiphoton excitation of fluorescent biomolecules like NAD(P)H, flavins, porphyrins, elastin, and melanin as well as the second harmonic generation of collagen is induced by picojoule femtosecond laser pulses from an tunable turn-key near infrared laser system. The ability for rapid highquality image acquisition, the user-friendly operation of the system, and the compact and flexible design qualifies this system to be used for melanoma detection, diagnostics of dermatological disorders, cosmetic research, and skin aging measurements as well as in situ drug monitoring and animal research. So far, more than 1,000 patients and volunteers have been investigated with the multiphoton tomographs in Europe, Asia, and Australia.

  12. Multiphoton ionization of atomic cesium

    International Nuclear Information System (INIS)

    Compton, R.N.; Klots, C.E.; Stockdale, J.A.D.; Cooper, C.D.

    1984-01-01

    We describe experimental studies of resonantly enhanced multi-photon ionization (MPI) of cesium atoms in the presence and absence of an external electric field. In the zero-field studies, photo-electron angular distributions for one- and two-photon resonantly enhanced MPI are compared with the theory of Tang and Lambropoulos. Deviations of experiment from theory are attributed to hyperfine coupling effects in the resonant intermediate state. The agreement between theory and experiment is excellent. In the absence of an external electric field, signal due to two-photon resonant three-photon ionization of cesium via np states is undetectable. Application of an electric field mixes nearby nd and ns levels, thereby inducing excitation and subsequent ionization. Signal due to two-photon excitation of ns levels in field-free experiments is weak due to their small photoionization cross section. An electric field mixes nearby np levels which again allows detectable photo-ionization signal. For both ns and np states the ''field induced'' MPI signal increases as the square of the electric field for a given principal quantum number and increases rapidly with n for a given field strength

  13. Infrared multiphoton absorption and decomposition

    International Nuclear Information System (INIS)

    Evans, D.K.; McAlpine, R.D.

    1984-01-01

    The discovery of infrared laser induced multiphoton absorption (IRMPA) and decomposition (IRMPD) by Isenor and Richardson in 1971 generated a great deal of interest in these phenomena. This interest was increased with the discovery by Ambartzumian, Letokhov, Ryadbov and Chekalin that isotopically selective IRMPD was possible. One of the first speculations about these phenomena was that it might be possible to excite a particular mode of a molecule with the intense infrared laser beam and cause decomposition or chemical reaction by channels which do not predominate thermally, thus providing new synthetic routes for complex chemicals. The potential applications to isotope separation and novel chemistry stimulated efforts to understand the underlying physics and chemistry of these processes. At ICOMP I, in 1977 and at ICOMP II in 1980, several authors reviewed the current understandings of IRMPA and IRMPD as well as the particular aspect of isotope separation. There continues to be a great deal of effort into understanding IRMPA and IRMPD and we will briefly review some aspects of these efforts with particular emphasis on progress since ICOMP II. 31 references

  14. Multiphoton ionization of atomic cesium

    International Nuclear Information System (INIS)

    Compton, R.N.; Klots, C.E.; Stockdale, J.A.D.; Cooper, C.D.

    1984-01-01

    We describe experimental studies of resonantly enhanced multiphoton ionization (MPI) of cesium atoms in the presence and absence of an external electric field. In the zero-field studies, photoelectron angular distributions for one- and two-photon resonantly enhanced MPI are compared with the theory of Tang and Lambropoulos. Deviations of experiment from theory are attributed to hyperfine coupling effects in the resonant intermediate state. The agreement between theory and experiment is excellent. In the absence of an external electric field, signal due to two-photon resonant three-photon ionization of cesium via np states is undetectable. Application of an electric field mixes nearby nd and ns levels, thereby inducing excitation and subsequent ionization. Signal due to two-photon excitation of ns levels in field-free experiments is weak due to their small photoionization cross section. An electric field mixes nearby np levels which again allows detectable photoionization signal. For both ns and np states the field induced MPI signal increases as the square of the electric field for a given principal quantum number and increases rapidly with n for a given field strength. Finally, we note that the classical two-photon field-ionization threshold is lower for the case in which the laser polarization and the electric field are parallel than it is when they are perpendicular. 22 references, 11 figures

  15. Multiphoton states and amplitude k-th power squeezing

    International Nuclear Information System (INIS)

    Buzek, V.; Jex, I.

    1991-01-01

    On the basis of the work of d'Ariano and coworkers a new type of multiphoton states is introduced. Amplitude k-th power squeezing of the multiphoton states are analysed. In particular, it is shown that even if the multiphoton states do not exhibit ordinary squeezing they can be amplitude k-th power squeezed

  16. Dynamic multiphoton imaging of acellular dermal matrix scaffolds seeded with mesenchymal stem cells in diabetic wound healing.

    Science.gov (United States)

    Chu, Jing; Shi, Panpan; Deng, Xiaoyuan; Jin, Ying; Liu, Hao; Chen, Maosheng; Han, Xue; Liu, Hanping

    2018-03-25

    Significantly effective therapies need to be developed for chronic nonhealing diabetic wounds. In this work, the topical transplantation of mesenchymal stem cell (MSC) seeded on an acellular dermal matrix (ADM) scaffold is proposed as a novel therapeutic strategy for diabetic cutaneous wound healing. GFP-labeled MSCs were cocultured with an ADM scaffold that was decellularized from normal mouse skin. These cultures were subsequently transplanted as a whole into the full-thickness cutaneous wound site in streptozotocin-induced diabetic mice. Wounds treated with MSC-ADM demonstrated an increased percentage of wound closure. The treatment of MSC-ADM also greatly increased angiogenesis and rapidly completed the reepithelialization of newly formed skin on diabetic mice. More importantly, multiphoton microscopy was used for the intravital and dynamic monitoring of collagen type I (Col-I) fibers synthesis via second harmonic generation imaging. The synthesis of Col-I fibers during diabetic wound healing is of great significance for revealing wound repair mechanisms. In addition, the activity of GFP-labeled MSCs during wound healing was simultaneously traced via two-photon excitation fluorescence imaging. Our research offers a novel advanced nonlinear optical imaging method for monitoring the diabetic wound healing process while the ADM and MSCs interact in situ. Schematic of dynamic imaging of ADM scaffolds seeded with mesenchymal stem cells in diabetic wound healing using multiphoton microscopy. PMT, photo-multiplier tube. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. A pragmatic guide to multiphoton microscope design

    Science.gov (United States)

    Young, Michael D.; Field, Jeffrey J.; Sheetz, Kraig E.; Bartels, Randy A.; Squier, Jeff

    2016-01-01

    Multiphoton microscopy has emerged as a ubiquitous tool for studying microscopic structure and function across a broad range of disciplines. As such, the intent of this paper is to present a comprehensive resource for the construction and performance evaluation of a multiphoton microscope that will be understandable to the broad range of scientific fields that presently exploit, or wish to begin exploiting, this powerful technology. With this in mind, we have developed a guide to aid in the design of a multiphoton microscope. We discuss source selection, optical management of dispersion, image-relay systems with scan optics, objective-lens selection, single-element light-collection theory, photon-counting detection, image rendering, and finally, an illustrated guide for building an example microscope. PMID:27182429

  18. Design and development of compact multiphoton microscopes

    Science.gov (United States)

    Mehravar, SeyedSoroush

    A compact multi-photon microscope (MPM) was designed and developed with the use of low-cost mode-locked fiber lasers operating at 1040nm and 1560nm. The MPM was assembled in-house and the system aberration was investigated using the optical design software: Zemax. A novel characterization methodology based on 'nonlinear knife-edge' technique was also introduced to measure the axial, lateral resolution, and the field curvature of the multi-photon microscope's image plane. The field curvature was then post-corrected using data processing in MATLAB. A customized laser scanning software based on LabVIEW was developed for data acquisition, image display and controlling peripheral electronics. Finally, different modalities of multi-photon excitation such as second- and third harmonic generation, two- and three-photon fluorescence were utilized to study a wide variety of samples from cancerous cells to 2D-layered materials.

  19. From morphology to clinical pathophysiology: multiphoton fluorescence lifetime imaging at patients' bedside

    Science.gov (United States)

    Mess, Christian; Zens, Katharina; Gorzelanny, Christian; Metze, Dieter; Luger, Thomas A.; König, Karsten; Schneider, Stefan W.; Huck, Volker

    2017-02-01

    Application of multiphoton microscopy in the field of biomedical research and advanced diagnostics promises unique insights into the pathophysiology of skin diseases. By means of multiphoton excitation, endogenous biomolecules like NADH, collagen or elastin show autofluorescence or second harmonic generation. Thus, these molecules provide information about the subcellular morphology, epidermal architecture and physiological condition of the skin. To gain a deeper understanding of the linkage between cellular structure and physiological processes, non-invasive multiphotonbased intravital tomography (MPT) and fluorescence lifetime imaging (FLIM) were combined within the scopes of inflammatory skin, chronic wounds and drug delivery in clinical application. The optical biopsies generated via MPT were morphologically analyzed and aligned with classical skin histology. Because of its subcellular resolution, MPT provided evidence of a redistribution of mitochondria in keratinocytes, indicating an altered cellular metabolism. Independent morphometric algorithms reliably showed a perinuclear accumulation in lesional skin in contrast to an even distribution in healthy skin. Confirmatively, MPT-FLIM showed an obvious metabolic shift in lesions. Moreover, detection of the onset and progression of inflammatory processes could be achieved. The feasibility of primary in vivo tracking of applied therapeutic agents further broadened our scope: We examined the permeation and subsequent distribution of agents directly visualized in patientś skin in short-term repetitive measurements. Furthermore, we performed MPT-FLIM follow-up investigations in the long-term course of therapy. Therefore, clinical MPT-FLIM application offers new insights into the pathophysiology and the individual therapeutic course of skin diseases, facilitating a better understanding of the processes of inflammation and wound healing.

  20. New developments in multimodal clinical multiphoton tomography

    Science.gov (United States)

    König, Karsten

    2011-03-01

    80 years ago, the PhD student Maria Goeppert predicted in her thesis in Goettingen, Germany, two-photon effects. It took 30 years to prove her theory, and another three decades to realize the first two-photon microscope. With the beginning of this millennium, first clinical multiphoton tomographs started operation in research institutions, hospitals, and in the cosmetic industry. The multiphoton tomograph MPTflexTM with its miniaturized flexible scan head became the Prism-Award 2010 winner in the category Life Sciences. Multiphoton tomographs with its superior submicron spatial resolution can be upgraded to 5D imaging tools by adding spectral time-correlated single photon counting units. Furthermore, multimodal hybrid tomographs provide chemical fingerprinting and fast wide-field imaging. The world's first clinical CARS studies have been performed with a hybrid multimodal multiphoton tomograph in spring 2010. In particular, nonfluorescent lipids and water as well as mitochondrial fluorescent NAD(P)H, fluorescent elastin, keratin, and melanin as well as SHG-active collagen have been imaged in patients with dermatological disorders. Further multimodal approaches include the combination of multiphoton tomographs with low-resolution imaging tools such as ultrasound, optoacoustic, OCT, and dermoscopy systems. Multiphoton tomographs are currently employed in Australia, Japan, the US, and in several European countries for early diagnosis of skin cancer (malignant melanoma), optimization of treatment strategies (wound healing, dermatitis), and cosmetic research including long-term biosafety tests of ZnO sunscreen nanoparticles and the measurement of the stimulated biosynthesis of collagen by anti-ageing products.

  1. Human bladder cancer diagnosis using multiphoton microscopy

    Science.gov (United States)

    Mukherjee, Sushmita; Wysock, James S.; Ng, Casey K.; Akhtar, Mohammed; Perner, Sven; Lee, Ming-Ming; Rubin, Mark A.; Maxfield, Frederick R.; Webb, Watt W.; Scherr, Douglas S.

    2009-02-01

    At the time of diagnosis, approximately 75% of bladder cancers are non-muscle invasive. Appropriate diagnosis and surgical resection at this stage improves prognosis dramatically. However, these lesions, being small and/or flat, are often missed by conventional white-light cystoscopes. Furthermore, it is difficult to assess the surgical margin for negativity using conventional cystoscopes. Resultantly, the recurrence rates in patients with early bladder cancer are very high. This is currently addressed by repeat cystoscopies and biopsies, which can last throughout the life of a patient, increasing cost and patient morbidity. Multiphoton endoscopes offer a potential solution, allowing real time, noninvasive biopsies of the human bladder, as well as an up-close assessment of the resection margin. While miniaturization of the Multiphoton microscope into an endoscopic format is currently in progress, we present results here indicating that Multiphoton imaging (using a bench-top Multiphoton microscope) can indeed identify cancers in fresh, unfixed human bladder biopsies. Multiphoton images are acquired in two channels: (1) broadband autofluorescence from cells, and (2) second harmonic generation (SHG), mostly by tissue collagen. These images are then compared with gold standard hematoxylin/eosin (H&E) stained histopathology slides from the same specimen. Based on a "training set" and a very small "blinded set" of samples, we have found excellent correlation between the Multiphoton and histopathological diagnoses. A larger blinded analysis by two independent uropathologists is currently in progress. We expect that the conclusion of this phase will provide us with diagnostic accuracy estimates, as well as the degree of inter-observer heterogeneity.

  2. Structure of multiphoton quantum optics. II. Bipartite systems, physical processes, and heterodyne squeezed states

    Science.gov (United States)

    dell'Anno, Fabio; de Siena, Silvio; Illuminati, Fabrizio

    2004-03-01

    Extending the scheme developed for a single mode of the electromagnetic field in the preceding paper [F. Dell’Anno, S. De Siena, and F. Illuminati, Phys. Rev. A 69, 033812 (2004)], we introduce two-mode nonlinear canonical transformations depending on two heterodyne mixing angles. They are defined in terms of Hermitian nonlinear functions that realize heterodyne superpositions of conjugate quadratures of bipartite systems. The canonical transformations diagonalize a class of Hamiltonians describing nondegenerate and degenerate multiphoton processes. We determine the coherent states associated with the canonical transformations, which generalize the nondegenerate two-photon squeezed states. Such heterodyne multiphoton squeezed states are defined as the simultaneous eigenstates of the transformed, coupled annihilation operators. They are generated by nonlinear unitary evolutions acting on two-mode squeezed states. They are non-Gaussian, highly nonclassical, entangled states. For a quadratic nonlinearity the heterodyne multiphoton squeezed states define two-mode cubic phase states. The statistical properties of these states can be widely adjusted by tuning the heterodyne mixing angles, the phases of the nonlinear couplings, as well as the strength of the nonlinearity. For quadratic nonlinearity, we study the higher-order contributions to the susceptibility in nonlinear media and we suggest possible experimental realizations of multiphoton conversion processes generating the cubic-phase heterodyne squeezed states.

  3. Structure of multiphoton quantum optics. II. Bipartite systems, physical processes, and heterodyne squeezed states

    International Nuclear Information System (INIS)

    Dell'Anno, Fabio; De Siena, Silvio; Illuminati, Fabrizio

    2004-01-01

    Extending the scheme developed for a single mode of the electromagnetic field in the preceding paper [F. Dell'Anno, S. De Siena, and F. Illuminati, Phys. Rev. A 69, 033812 (2004)], we introduce two-mode nonlinear canonical transformations depending on two heterodyne mixing angles. They are defined in terms of Hermitian nonlinear functions that realize heterodyne superpositions of conjugate quadratures of bipartite systems. The canonical transformations diagonalize a class of Hamiltonians describing nondegenerate and degenerate multiphoton processes. We determine the coherent states associated with the canonical transformations, which generalize the nondegenerate two-photon squeezed states. Such heterodyne multiphoton squeezed states are defined as the simultaneous eigenstates of the transformed, coupled annihilation operators. They are generated by nonlinear unitary evolutions acting on two-mode squeezed states. They are non-Gaussian, highly nonclassical, entangled states. For a quadratic nonlinearity the heterodyne multiphoton squeezed states define two-mode cubic phase states. The statistical properties of these states can be widely adjusted by tuning the heterodyne mixing angles, the phases of the nonlinear couplings, as well as the strength of the nonlinearity. For quadratic nonlinearity, we study the higher-order contributions to the susceptibility in nonlinear media and we suggest possible experimental realizations of multiphoton conversion processes generating the cubic-phase heterodyne squeezed states

  4. Theory of multiphoton ionization of atoms

    International Nuclear Information System (INIS)

    Szoeke, A.

    1986-03-01

    A non-perturbative approach to the theory of multiphoton ionization is reviewed. Adiabatic Floquet theory is its first approximation. It explains qualitatively the energy and angular distribution of photoelectrons. In many-electron atoms it predicts collective and inner shell excitation. 14 refs

  5. Thermostatic tissue platform for intravital microscopy: 'the hanging drop' model.

    Science.gov (United States)

    Pavlovic, Dragan; Frieling, Helge; Lauer, Kai-Stephan; Bac, Vo Hoai; Richter, Joern; Wendt, Michael; Lehmann, Christian; Usichenko, Taras; Meissner, Konrad; Gruendling, Matthias

    2006-11-01

    Intravital microscopy imposes the particular problem of the combined control of the body temperature of the animal and the local temperature of the observed organ or tissues. We constructed and tested, in the rat ileum microcirculation preparation, a new organ-support platform. The platform consisted of an organ bath filled with physiological solution, and contained a suction tube, a superfusion tube, an intestine-support hand that was attached to a micromanipulator and a thermometer probe. To cover the intestine we used a cover glass plate with a plastic ring glued on its upper surface. After a routine procedure (anaesthesia, monitoring and surgery), the intestine segment (2-3 cm long) was gently exteriorized and placed on the 'hand' of the organ support. A small part of the intestine formed a small 'island' in the bath that was filled with physiological salt solution. The cover glass was secured in place. The physiological salt solution from the superfusion tube, which was pointed to the lower surface of the cover glass, formed a 'hanging drop'. The objective of the microscope was then immersed into distilled water that was formed by the cover glass plastic ring. The 'hanging drop' technique prevented any tissue quenching, ensured undisturbed microcirculation, provided for stable temperature and humidity, and permitted a clear visual field.

  6. Intravital Fluorescence Excitation in Whole-Animal Optical Imaging.

    Science.gov (United States)

    Nooshabadi, Fatemeh; Yang, Hee-Jeong; Bixler, Joel N; Kong, Ying; Cirillo, Jeffrey D; Maitland, Kristen C

    2016-01-01

    Whole-animal fluorescence imaging with recombinant or fluorescently-tagged pathogens or cells enables real-time analysis of disease progression and treatment response in live animals. Tissue absorption limits penetration of fluorescence excitation light, particularly in the visible wavelength range, resulting in reduced sensitivity to deep targets. Here, we demonstrate the use of an optical fiber bundle to deliver light into the mouse lung to excite fluorescent bacteria, circumventing tissue absorption of excitation light in whole-animal imaging. We present the use of this technology to improve detection of recombinant reporter strains of tdTomato-expressing Mycobacterium bovis BCG (Bacillus Calmette Guerin) bacteria in the mouse lung. A microendoscope was integrated into a whole-animal fluorescence imager to enable intravital excitation in the mouse lung with whole-animal detection. Using this technique, the threshold of detection was measured as 103 colony forming units (CFU) during pulmonary infection. In comparison, the threshold of detection for whole-animal fluorescence imaging using standard epi-illumination was greater than 106 CFU.

  7. Intravital imaging of CD8+ T cell function in cancer.

    Science.gov (United States)

    Mempel, Thorsten R; Bauer, Christian A

    2009-01-01

    Recent technological advances in photonics are making intravital microscopy (IVM) an increasingly powerful approach for the mechanistic exploration of biological processes in the physiological context of complex native tissue environments. Direct, dynamic and multiparametric visualization of immune cell behavior in living animals at cellular and subcellular resolution has already proved its utility in auditing basic immunological concepts established through conventional approaches and has also generated new hypotheses that can conversely be complemented and refined by traditional experimental methods. The insight that outgrowing tumors must not necessarily have evaded recognition by the adaptive immune system, but can escape rejection by actively inducing a state of immunological tolerance calls for a detailed investigation of the cellular and molecular mechanisms by which the anti-cancer response is subverted. Along with molecular imaging techniques that provide dynamic information at the population level, IVM can be expected to make a critical contribution to this effort by allowing the observation of immune cell behavior in vivo at single cell-resolution. We review here how IVM-based investigation can help to clarify the role of cytotoxic T lymphocytes (CTL) in the immune response against cancer and identify the ways by which their function might be impaired through tolerogenic mechanisms.

  8. Study the multi-photon absorption process in two types of molecules

    International Nuclear Information System (INIS)

    Al-azawi, H.R.

    1986-01-01

    The aim of the present work was to study the multi-photon absorption process in two types of molecules; spherical top such as SF 6 molecules and assymetric top such as CHOOH and C 2 H 4 molecules. This work also aimed to study the effect of buffer gas pressure (Ar), which is transparent to the infrared (IR) laser on the multiphoton absorption of both types of molecules. A pulsed (TEA) CO 2 laser was used as a source which generates multi-lines in the IR-region of the spectrum and an optoacoustic detector was used to detect the energy absorbed by the molecules. In this study, the relaxation process was found to be faster in the heavy molecules than that in the light ones. A limit in the Ar pressure was observed. Below this limit, the gas acted as an active buffer gas and above it, the multi-photon absorption process was quenched. This work also aimed to study the multi-photon absorption spectrum for the CHOOH molecules in the range (1067-1090 cm -1 ). This spectrum was found to be consistent with the linear absorption spectrum obtained for the same range. The density of the vibrational states as a function of the vibrational energy was studied for the molecules SF 6 , CHOOH and C 2 H 4 . The results were used to interpret (i) the difference in the energy absorbed by difference molecules at the same energy density and (ii) the non-linearity in the multi-photon absorption for CHOOH molecules. 1 tab.; 40 figs.; 70 refs

  9. Multiphoton tomography of intratissue tattoo nanoparticles

    Science.gov (United States)

    König, Karsten

    2012-02-01

    Most of today's intratissue tattoo pigments are unknown nanoparticles. So far, there was no real control of their use due to the absence of regulations. Some of the tattoo pigments contain carcinogenic amines e.g. azo pigment Red 22. Nowadays, the European Union starts to control the administration of tattoo pigments. There is an interest to obtain information on the intratissue distribution, their interaction with living cells and the extracellular matrix, and the mechanisms behind laser tattoo removal. Multiphoton tomographs are novel biosafety and imaging tools that can provide such information non-invasively and without further labeling. When using the spectral FLIM module, spatially-resolved emission spectra, excitation spectra, and fluorescence lifetimes can pr provided. Multiphoton tomographs are used by all major cosmetic comapanies to test the biosafety of sunscreen nanoparticles.

  10. Multiphoton ionization/dissociation of osmium tetroxide

    International Nuclear Information System (INIS)

    Ding, D.; Puretzky, A.A.; Compton, R.N.

    1993-01-01

    The mechanisms leading to laser multiphoton ionization and dissociation (MPI/MPD) of osmium tetroxide (OsO 4 ) have been investigated from measurements of the kinetic energies of product ions (Os + , Os 2+ , OsO + , O 2 + , O + ) and photoelectrons as a function of the laser wavelength. Neutral channels, intermediate to the dominant Os + ionization channel, such as OsO 4 →OsO 4-n +nO are examined using resonance-enhanced multiphoton ionization (REMPI) of the fast O atoms. Equipartition of the available photon energy among the fragments is observed. The wavelength dependence of the Os + ion signal suggests that one or more of the steps leading to Os + ions involve molecular ions and/or excited neutral atoms. The observed preponderance of very slow ( 2+ is shown to result primarily from REMPI of Os +

  11. Multiphoton tomography to detect chemo- and biohazards

    Science.gov (United States)

    König, Karsten

    2015-03-01

    In vivo high-resolution multiphoton/CARS tomography provides optical biopsies with 300 nm lateral resolution with chemical fingerprints. Thousands of volunteers and patients have been investigated for early cancer diagnosis, evaluation of anti-ageing cosmetic products, and changes of cellular metabolism by UV exposure and decreased oxygen supply. The skin as the outermost and largest organ is also the major target of CB agents. Current UV-based sensors are useful for bio-aerosol sensing but not for evaluating exposed in vivo skin. Here we evaluate the use of 4D multiphoton/CARS tomographs based on near infrared femtosecond laser radiation, time-correlated single photon counting (FLIM) and white light generation by photonic crystal fibers to detect bio- and chemohazards in human in vivo skin using twophoton fluorescence, SHG, and Raman signals.

  12. Multiphoton processes in isolated atoms and molecules

    International Nuclear Information System (INIS)

    Sudbo, A.S.

    1979-11-01

    The theory of coherent excitation of a multilevel quantum mechanical system is developed. Damping of the system is taken into account by the use of a density matrix formalism. General properties of the wave function and/or the density matrix are discussed. The physical implications for the behavior of the system are described, together with possible applications of the formalism, including the infrared multiphoton excitation of molecules, and optical pumping in alkali atoms. Experimental results are presented on the infrared multiphoton dissociation of molecules, followed by a discussion of the general features of this process. The experimental results were obtained using a crossed laser and molecular beam method, and the emphasis is on determining the properties of the dissociating molecule and the dissociation products. The dissociation process is shown to be described very well by the standard statistical theory (RRKM theory) of unimolecular reactions, a brief presentation of which is also included

  13. Multiphoton spectroscopy of human skin in vivo

    Science.gov (United States)

    Breunig, Hans G.; Weinigel, Martin; König, Karsten

    2012-03-01

    In vivo multiphoton-intensity images and emission spectra of human skin are reported. Optical sections from different depths of the epidermis and dermis have been measured with near-infrared laser-pulse excitation. While the intensity images reveal information on the morphology, the spectra show emission characteristics of main endogenous skin fluorophores like keratin, NAD(P)H, melanin, elastin and collagen as well as of second harmonic generation induced by the excitation-light interaction with the dermal collagen network.

  14. Dressing effect in multiphoton unimolecular dissociation

    Energy Technology Data Exchange (ETDEWEB)

    Gonzalez-Diaz, P.F.; Garcia-Fernandez, P.

    1986-03-01

    On the basis of a quantum-statistical model recently discussed, we deal in this paper with the perturbations induced by the intense field of a CO/sub 2/ laser on the levels of the vibrational pattern of a molecule undergoing multiphoton unimolecular dissociation. This perturbational correction is investigated by using a displacement operator technique and the results are interpreted according to the statistical model.

  15. Multiphoton ionization processes in strong laser

    International Nuclear Information System (INIS)

    Krstic, P.

    1982-01-01

    Multiphoton ionization of hydrogen in ultrastrong laser fields is studied. The previous calculations of this process yield differing result for the transition rate. We show the relations between them and difficulties with each of them. One difficulty is that the finite spatial and time extent of the laser field has been omitted. It is also found that a laser field, which is sufficiently intense to be labeled ultrastrong, makes the electron move relativistically so that it becomes necessary to use Volkov states to describe the electron in the laser field. The transition rate is obtained, using a CO laser as an example, and it is found that the transition rate rises as the laser intensity rises. This is a consequence of the use of relativistic kinematics and is not true nonrelativistically. We also discuss the multiple peaks observed in the energy spectrum of electrons resulting from multiphoton ionization of atoms by lasers. When the laser intensity is large enough for the ponderomotive force to result in appreciable broading of the peaks we show the shape of the broadened peaks contains useful information. We show that the multiphoton ionization probability as a function of laser intensity can be obtained but that the free-free cross sections, which are in principle also obtainable, are probably not obtainable in practice. Finally, we describe the theory of the absorption of more than minimum numbers of photons needed to ionize an atom by an intense laser. The basic approximation used is that the atom is adiabatically deformed by the laser and an impulsive interaction then results in multiphoton absorption. In our first calculation we allow only one resonant excited state to be included in the adiabatic deformation. In our second we also allow the lowest energy continuum to be included. The two results are then compared

  16. Fundamental studies of molecular multiphoton ionization

    International Nuclear Information System (INIS)

    Miller, J.C.; Compton, R.N.

    1984-04-01

    For several years the authors have performed fundamental studies of molecular multiphoton ionization (MPI). We will present a potpourri of techniques and results chosen to illustrate the interesting complexities of molecular MPI. Techniques used include time-of-flight mass spectroscopy, photoelectron spectroscopy, supersonic expansion cooling of molecular beams, harmonic generation, two-color laser MPI, and polarization spectroscopy. Whenever possible the relevance of these results to resonance ionization spectroscopy schemes will be delineated. 23 references, 10 figures

  17. Multiphotonic resonance processes in potassium vapor

    International Nuclear Information System (INIS)

    Bensoussan, Paul.

    1975-01-01

    Despite several theoretical and experimental investigations, the phenomena of resonance multiphotonic ionization are still not completely understood. The following lines of investigation were undertaken to try and elucidate certain aspects of the resonance processes. The first line of investigation aims at finding the processes which can compete with ionization. Resonance ionization processes can be considered as taking place in two stages: absorption induced excitation of a bound state, followed by photoionization from the excited level. The problem is now to determine what are the processes which compete with the ionization processes starting from a level selectively populated by the absorption of one or two photons. The second line aims at finding the influence of the polarization of the radiation on resonance multiphotonic ionization for the second photon and to check the validity of the selection rules on the magnetic quantic number of the resonance bound linked states. The last study therefore relates to the development of a method of multiphotonic spectrometry which could determine the energy levels in the alcaline f series [fr

  18. Spin Multiphoton Antiresonance at Finite Temperatures

    Science.gov (United States)

    Hicke, Christian; Dykman, Mark

    2007-03-01

    Weakly anisotropic S>1 spin systems display multiphoton antiresonance. It occurs when an Nth overtone of the radiation frequency coincides with the distance between the ground and the Nth excited energy level (divided by ). The coherent response of the spin displays a sharp minimum or maximum as a function of frequency, depending on which state was initially occupied. We find the spectral shape of the response dips/peaks. We also study the stationary response for zero and finite temperatures. The response changes dramatically with increasing temperature, when excited states become occupied even in the absence of radiation. The change is due primarily to the increasing role of single-photon resonances between excited states, which occur at the same frequencies as multiphoton resonances. Single-photon resonances are broad, because the single-photon Rabi frequencies largely exceed the multi-photon ones. This allows us to separate different resonances and to study their spectral shape. We also study the change of the spectrum due to relaxational broadening of the peaks, with account taken of both decay and phase modulation.

  19. From good to bad : Intravital imaging of the hijack of physiological processes by cancer cells

    NARCIS (Netherlands)

    Suijkerbuijk, Saskia J.E.; van Rheenen, Jacco

    2017-01-01

    Homeostasis of tissues is tightly regulated at the cellular, tissue and organismal level. Interestingly, tumor cells have found ways to hijack many of these physiological processes at all the different levels. Here we review how intravital microscopy techniques have provided new insights into our

  20. Multi-photon excited luminescence of magnetic FePt core-shell nanoparticles.

    Science.gov (United States)

    Seemann, K M; Kuhn, B

    2014-07-01

    We present magnetic FePt nanoparticles with a hydrophilic, inert, and biocompatible silico-tungsten oxide shell. The particles can be functionalized, optically detected, and optically manipulated. To show the functionalization the fluorescent dye NOPS was bound to the FePt core-shell nanoparticles with propyl-triethoxy-silane linkers and fluorescence of the labeled particles were observed in ethanol (EtOH). In aqueous dispersion the NOPS fluorescence is quenched making them invisible using 1-photon excitation. However, we observe bright luminescence of labeled and even unlabeled magnetic core-shell nanoparticles with multi-photon excitation. Luminescence can be detected in the near ultraviolet and the full visible spectral range by near infrared multi-photon excitation. For optical manipulation, we were able to drag clusters of particles, and maybe also single particles, by a focused laser beam that acts as optical tweezers by inducing an electric dipole in the insulated metal nanoparticles. In a first application, we show that the luminescence of the core-shell nanoparticles is bright enough for in vivo multi-photon imaging in the mouse neocortex down to cortical layer 5.

  1. Ponderomotive effects in multiphoton pair production

    Science.gov (United States)

    Kohlfürst, Christian; Alkofer, Reinhard

    2018-02-01

    The Dirac-Heisenberg-Wigner formalism is employed to investigate electron-positron pair production in cylindrically symmetric but otherwise spatially inhomogeneous, oscillating electric fields. The oscillation frequencies are hereby tuned to obtain multiphoton pair production in the nonperturbative threshold regime. An effective mass, as well as a trajectory-based semiclassical analysis, is introduced in order to interpret the numerical results for the distribution functions as well as for the particle yields and spectra. The results, including the asymptotic particle spectra, display clear signatures of ponderomotive forces.

  2. Multiphoton tomography of the human eye

    Science.gov (United States)

    König, Karsten; Batista, Ana; Hager, Tobias; Seitz, Berthold

    2017-02-01

    Multiphoton tomography (MPT) is a novel label-free clinical imaging method for non-invasive tissue imaging with high spatial (300 nm) and temporal (100 ps) resolutions. In vivo optical histology can be realized due to the nonlinear excitation of endogenous fluorophores and second-harmonic generation (SHG) of collagen. Furthermore, optical metabolic imaging (OMI) is performed by two-photon autofluorescence lifetime imaging (FLIM). So far, applications of the multiphoton tomographs DermaInspect and MPTflex were limited to dermatology. Novel applications include intraoperative brain tumor imaging as well as cornea imaging. In this work we describe two-photon imaging of ex vivo human corneas unsuitable for transplantation. Furthermore, the cross-linking (CXL) process of corneal collagen based on UVA exposure and 0.1 % riboflavin was studied. The pharmacokinetics of the photosensitizer could be detected with high spatial resolution. Interestingly, an increase in the stromal autofluorescence intensity and modifications of the autofluorescence lifetimes were observed in the human corneal samples within a few days following CXL.

  3. Efficient Multiphoton Generation in Waveguide Quantum Electrodynamics

    Science.gov (United States)

    González-Tudela, A.; Paulisch, V.; Kimble, H. J.; Cirac, J. I.

    2017-05-01

    Engineering quantum states of light is at the basis of many quantum technologies such as quantum cryptography, teleportation, or metrology among others. Though, single photons can be generated in many scenarios, the efficient and reliable generation of complex single-mode multiphoton states is still a long-standing goal in the field, as current methods either suffer from low fidelities or small probabilities. Here we discuss several protocols which harness the strong and long-range atomic interactions induced by waveguide QED to efficiently load excitations in a collection of atoms, which can then be triggered to produce the desired multiphoton state. In order to boost the success probability and fidelity of each excitation process, atoms are used to both generate the excitations in the rest, as well as to herald the successful generation. Furthermore, to overcome the exponential scaling of the probability of success with the number of excitations, we design a protocol to merge excitations that are present in different internal atomic levels with a polynomial scaling.

  4. Multifocal multiphoton microscopy with adaptive optical correction

    Science.gov (United States)

    Coelho, Simao; Poland, Simon; Krstajic, Nikola; Li, David; Monypenny, James; Walker, Richard; Tyndall, David; Ng, Tony; Henderson, Robert; Ameer-Beg, Simon

    2013-02-01

    Fluorescence lifetime imaging microscopy (FLIM) is a well established approach for measuring dynamic signalling events inside living cells, including detection of protein-protein interactions. The improvement in optical penetration of infrared light compared with linear excitation due to Rayleigh scattering and low absorption have provided imaging depths of up to 1mm in brain tissue but significant image degradation occurs as samples distort (aberrate) the infrared excitation beam. Multiphoton time-correlated single photon counting (TCSPC) FLIM is a method for obtaining functional, high resolution images of biological structures. In order to achieve good statistical accuracy TCSPC typically requires long acquisition times. We report the development of a multifocal multiphoton microscope (MMM), titled MegaFLI. Beam parallelization performed via a 3D Gerchberg-Saxton (GS) algorithm using a Spatial Light Modulator (SLM), increases TCSPC count rate proportional to the number of beamlets produced. A weighted 3D GS algorithm is employed to improve homogeneity. An added benefit is the implementation of flexible and adaptive optical correction. Adaptive optics performed by means of Zernike polynomials are used to correct for system induced aberrations. Here we present results with significant improvement in throughput obtained using a novel complementary metal-oxide-semiconductor (CMOS) 1024 pixel single-photon avalanche diode (SPAD) array, opening the way to truly high-throughput FLIM.

  5. Multiphoton excitation and high-harmonics generation in topological insulator.

    Science.gov (United States)

    Avetissian, H K; Avetissian, A K; Avchyan, B R; Mkrtchian, G F

    2018-05-10

    Multiphoton interaction of coherent electromagnetic radiation with 2D metallic carriers confined on the surface of the 3D topological insulator is considered. A microscopic theory describing the nonlinear interaction of a strong wave and metallic carriers with many-body Coulomb interaction is developed. The set of integrodifferential equations for the interband polarization and carrier occupation distribution is solved numerically. Multiphoton excitation of Fermi-Dirac sea of 2D massless carriers is considered for a THz pump wave. It is shown that in the moderately strong pump wave field along with multiphoton interband/intraband transitions the intense radiation of high harmonics takes place.

  6. QED theory of multiphoton transitions in atoms and ions

    Science.gov (United States)

    Zalialiutdinov, Timur A.; Solovyev, Dmitry A.; Labzowsky, Leonti N.; Plunien, Günter

    2018-03-01

    This review surveys the quantum theory of electromagnetic radiation for atomic systems. In particular, a review of current theoretical studies of multiphoton processes in one and two-electron atoms and highly charged ions is provided. Grounded on the quantum electrodynamics description the multiphoton transitions in presence of cascades, spin-statistic behaviour of equivalent photons and influence of external electric fields on multiphoton in atoms and anti-atoms are discussed. Finally, the nonresonant corrections which define the validity of the concept of the excited state energy levels are introduced.

  7. Multiphoton excitation and high-harmonics generation in topological insulator

    Science.gov (United States)

    Avetissian, H. K.; Avetissian, A. K.; Avchyan, B. R.; Mkrtchian, G. F.

    2018-05-01

    Multiphoton interaction of coherent electromagnetic radiation with 2D metallic carriers confined on the surface of the 3D topological insulator is considered. A microscopic theory describing the nonlinear interaction of a strong wave and metallic carriers with many-body Coulomb interaction is developed. The set of integrodifferential equations for the interband polarization and carrier occupation distribution is solved numerically. Multiphoton excitation of Fermi–Dirac sea of 2D massless carriers is considered for a THz pump wave. It is shown that in the moderately strong pump wave field along with multiphoton interband/intraband transitions the intense radiation of high harmonics takes place.

  8. Amplitudes for multiphoton quantum processes in linear optics

    International Nuclear Information System (INIS)

    UrIas, Jesus

    2011-01-01

    The prominent role that linear optical networks have acquired in the engineering of photon states calls for physically intuitive and automatic methods to compute the probability amplitudes for the multiphoton quantum processes occurring in linear optics. A version of Wick's theorem for the expectation value, on any vector state, of products of linear operators, in general, is proved. We use it to extract the combinatorics of any multiphoton quantum processes in linear optics. The result is presented as a concise rule to write down directly explicit formulae for the probability amplitude of any multiphoton process in linear optics. The rule achieves a considerable simplification and provides an intuitive physical insight about quantum multiphoton processes. The methodology is applied to the generation of high-photon-number entangled states by interferometrically mixing coherent light with spontaneously down-converted light.

  9. Amplitudes for multiphoton quantum processes in linear optics

    Science.gov (United States)

    Urías, Jesús

    2011-07-01

    The prominent role that linear optical networks have acquired in the engineering of photon states calls for physically intuitive and automatic methods to compute the probability amplitudes for the multiphoton quantum processes occurring in linear optics. A version of Wick's theorem for the expectation value, on any vector state, of products of linear operators, in general, is proved. We use it to extract the combinatorics of any multiphoton quantum processes in linear optics. The result is presented as a concise rule to write down directly explicit formulae for the probability amplitude of any multiphoton process in linear optics. The rule achieves a considerable simplification and provides an intuitive physical insight about quantum multiphoton processes. The methodology is applied to the generation of high-photon-number entangled states by interferometrically mixing coherent light with spontaneously down-converted light.

  10. Real-time intravital imaging of pH variation associated with osteoclast activity.

    Science.gov (United States)

    Maeda, Hiroki; Kowada, Toshiyuki; Kikuta, Junichi; Furuya, Masayuki; Shirazaki, Mai; Mizukami, Shin; Ishii, Masaru; Kikuchi, Kazuya

    2016-08-01

    Intravital imaging by two-photon excitation microscopy (TPEM) has been widely used to visualize cell functions. However, small molecular probes (SMPs), commonly used for cell imaging, cannot be simply applied to intravital imaging because of the challenge of delivering them into target tissues, as well as their undesirable physicochemical properties for TPEM imaging. Here, we designed and developed a functional SMP with an active-targeting moiety, higher photostability, and a fluorescence switch and then imaged target cell activity by injecting the SMP into living mice. The combination of the rationally designed SMP with a fluorescent protein as a reporter of cell localization enabled quantitation of osteoclast activity and time-lapse imaging of its in vivo function associated with changes in cell deformation and membrane fluctuations. Real-time imaging revealed heterogenic behaviors of osteoclasts in vivo and provided insights into the mechanism of bone resorption.

  11. Rotational averaging of multiphoton absorption cross sections

    Energy Technology Data Exchange (ETDEWEB)

    Friese, Daniel H., E-mail: daniel.h.friese@uit.no; Beerepoot, Maarten T. P.; Ruud, Kenneth [Centre for Theoretical and Computational Chemistry, University of Tromsø — The Arctic University of Norway, N-9037 Tromsø (Norway)

    2014-11-28

    Rotational averaging of tensors is a crucial step in the calculation of molecular properties in isotropic media. We present a scheme for the rotational averaging of multiphoton absorption cross sections. We extend existing literature on rotational averaging to even-rank tensors of arbitrary order and derive equations that require only the number of photons as input. In particular, we derive the first explicit expressions for the rotational average of five-, six-, and seven-photon absorption cross sections. This work is one of the required steps in making the calculation of these higher-order absorption properties possible. The results can be applied to any even-rank tensor provided linearly polarized light is used.

  12. Multiphoton amplitude in a constant background field

    Science.gov (United States)

    Ahmad, Aftab; Ahmadiniaz, Naser; Corradini, Olindo; Kim, Sang Pyo; Schubert, Christian

    2018-01-01

    In this contribution, we present our recent compact master formulas for the multiphoton amplitudes of a scalar propagator in a constant background field using the worldline fomulation of quantum field theory. The constant field has been included nonperturbatively, which is crucial for strong external fields. A possible application is the scattering of photons by electrons in a strong magnetic field, a process that has been a subject of great interest since the discovery of astrophysical objects like radio pulsars, which provide evidence that magnetic fields of the order of 1012G are present in nature. The presence of a strong external field leads to a strong deviation from the classical scattering amplitudes. We explicitly work out the Compton scattering amplitude in a magnetic field, which is a process of potential relevance for astrophysics. Our final result is compact and suitable for numerical integration.

  13. Limiting energy loss distributions for multiphoton channeling radiation

    International Nuclear Information System (INIS)

    Bondarenco, M.V.

    2015-01-01

    Recent results in the theory of multiphoton spectra for coherent radiation sources are overviewed, with the emphasis on channeling radiation. For the latter case, the importance of the order of resummation and averaging is emphasized. Limiting shapes of multiphoton spectra at high intensity are discussed for different channeling regimes. In some spectral regions, there emerges a correspondence between the radiative energy loss and the electron integrals of motion

  14. Studies of atmospheric molecules by multiphoton spectroscopy

    International Nuclear Information System (INIS)

    Johnson, P.M.

    1990-12-01

    Resonance ionization processes can play an important role in understanding molecules important in combustion processes. They are a reflection of the dynamic as well as the static properties of atomic and molecular species. Due to the sequential or quasisequential nature of photon absorption in resonant multiphoton events, the lifetimes of the intermediate states play an essential role in the overall cross-sections if they are short enough to be competitive with subsequent photon interactions. In molecules this is particularly important because there are many dissociative and other radiationless pathways which can contribute to a competitive channel. Under those conditions it should be possible to obtain information about the nature of the dynamics of the intermediate state form the multiphoton ionization process. This will involve looking at not only the ionization cross-section but also other observables such as the kinetic energy of the ejected electrons and possibly the distribution of fragment ions produced in the ionization event. Whether the ionization amplitude is affected or not, the time scales of the dynamic events which alter the ionization path can vary over a large range from the femtoseconds of dissociation to the microseconds of some radiationless transitions in large molecules. When the competing channel has a time scale shorter than the laser pulse length, the kinetics of the ionization are intimately tied into the precise nature of the laser pulse. For time scales longer than the laser pulse, pump-probe ionization schemes in which one laser prepares a state while another does the ionization provide a particularly simple method for investigating the dynamics of the intermediate state. Here the author discusses examples from each of these regimes. CO 2 and pyrazine are examined. 6 refs., 6 figs

  15. [Frontiers in Live Bone Imaging Researches. Novel drug discovery by means of intravital bone imaging technology].

    Science.gov (United States)

    Ishii, Masaru

    2015-06-01

    Recent advances in intravital bone imaging technology has enabled us to grasp the real cellular behaviors and functions in vivo , revolutionizing the field of drug discovery for novel therapeutics against intractable bone diseases. In this chapter, I introduce various updated information on pharmacological actions of several antibone resorptive agents, which could only be derived from advanced imaging techniques, and also discuss the future perspectives of this new trend in drug discovery.

  16. Intravital two-photon microscopy of immune cell dynamics in corneal lymphatic vessels.

    Directory of Open Access Journals (Sweden)

    Philipp Steven

    Full Text Available BACKGROUND: The role of lymphatic vessels in tissue and organ transplantation as well as in tumor growth and metastasis has drawn great attention in recent years. METHODOLOGY/PRINCIPAL FINDINGS: We now developed a novel method using non-invasive two-photon microscopy to simultaneously visualize and track specifically stained lymphatic vessels and autofluorescent adjacent tissues such as collagen fibrils, blood vessels and immune cells in the mouse model of corneal neovascularization in vivo. The mouse cornea serves as an ideal tissue for this technique due to its easy accessibility and its inducible and modifiable state of pathological hem- and lymphvascularization. Neovascularization was induced by suture placement in corneas of Balb/C mice. Two weeks after treatment, lymphatic vessels were stained intravital by intrastromal injection of a fluorescently labeled LYVE-1 antibody and the corneas were evaluated in vivo by two-photon microscopy (TPM. Intravital TPM was performed at 710 nm and 826 nm excitation wavelengths to detect immunofluorescence and tissue autofluorescence using a custom made animal holder. Corneas were then harvested, fixed and analyzed by histology. Time lapse imaging demonstrated the first in vivo evidence of immune cell migration into lymphatic vessels and luminal transport of individual cells. Cells immigrated within 1-5.5 min into the vessel lumen. Mean velocities of intrastromal corneal immune cells were around 9 µm/min and therefore comparable to those of T-cells and macrophages in other mucosal surfaces. CONCLUSIONS: To our knowledge we here demonstrate for the first time the intravital real-time transmigration of immune cells into lymphatic vessels. Overall this study demonstrates the valuable use of intravital autofluorescence two-photon microscopy in the model of suture-induced corneal vascularizations to study interactions of immune and subsequently tumor cells with lymphatic vessels under close as possible

  17. High-resolution multimodal clinical multiphoton tomography of skin

    Science.gov (United States)

    König, Karsten

    2011-03-01

    This review focuses on multimodal multiphoton tomography based on near infrared femtosecond lasers. Clinical multiphoton tomographs for 3D high-resolution in vivo imaging have been placed into the market several years ago. The second generation of this Prism-Award winning High-Tech skin imaging tool (MPTflex) was introduced in 2010. The same year, the world's first clinical CARS studies have been performed with a hybrid multimodal multiphoton tomograph. In particular, non-fluorescent lipids and water as well as mitochondrial fluorescent NAD(P)H, fluorescent elastin, keratin, and melanin as well as SHG-active collagen has been imaged with submicron resolution in patients suffering from psoriasis. Further multimodal approaches include the combination of multiphoton tomographs with low-resolution wide-field systems such as ultrasound, optoacoustical, OCT, and dermoscopy systems. Multiphoton tomographs are currently employed in Australia, Japan, the US, and in several European countries for early diagnosis of skin cancer, optimization of treatment strategies, and cosmetic research including long-term testing of sunscreen nanoparticles as well as anti-aging products.

  18. The multiphoton ionization of uranium hexafluoride

    International Nuclear Information System (INIS)

    Armstrong, D.P.

    1992-05-01

    Multiphoton ionization (MPI) time-of-flight mass spectroscopy and photoelectron spectroscopy studies of UF 6 have been conducted using focused light from the Nd:YAG laser fundamental (λ=1064 nm) and its harmonics (λ=532, 355, or 266 nm), as well as other wavelengths provided by a tunable dye laser. The MPI mass spectra are dominated by the singly and multiply charged uranium ions rather than by the UF x + fragment ions even at the lowest laser power densities at which signal could be detected. The laser power dependence of U n+ ions signals indicates that saturation can occur for many of the steps required for their ionization. In general, the doubly-charged uranium ion (U 2+ ) intensity is much greater than that of the singly-charged uranium ion (U + ). For the case of the tunable dye laser experiments, the U n+ (n = 1- 4) wavelength dependence is relatively unstructured and does not show observable resonance enhancement at known atomic uranium excitation wavelengths. The dominance of the U 2+ ion and the absence or very small intensities of UF x + fragments, along with the unsaturated wavelength dependence, indicate that mechanisms may exist other than ionization of bare U atoms after the stepwise photodissociation of F atoms from the parent molecule

  19. Evaluation of multiphoton effects in down-conversion

    International Nuclear Information System (INIS)

    Yoshimi, Kazuyoshi; Koshino, Kazuki

    2010-01-01

    Multiphoton effects in down-conversion are investigated based on the full-quantum multimode formalism by considering a three-level system as a prototype nonlinear system. We analytically derive the three-photon output wave function for two input photons, where one of the two input photons is down-converted and the other one is not. Using this output wave function, we calculate the down-conversion probability, the purity, and the fidelity to evaluate the entanglement between a down-converted photon pair and a non-down-converted photon. It is shown that the saturation effect occurs by multiphoton input and that it affects both the down-conversion probability and the quantum correlation between the down-converted photon pair and the non-down-converted photon. We also reveal the necessary conditions for multiphoton effects to be strong.

  20. A review of biomedical multiphoton microscopy and its laser sources

    International Nuclear Information System (INIS)

    Lefort, Claire

    2017-01-01

    Multiphoton microscopy (MPM) has been the subject of major development efforts for about 25 years for imaging biological specimens at micron scale and presented as an elegant alternative to classical fluorescence methods such as confocal microscopy. In this topical review, the main interests and technical requirements of MPM are addressed with a focus on the crucial role of excitation source for optimization of multiphoton processes. Then, an overview of the different sources successfully demonstrated in literature for MPM is presented, and their physical parameters are inventoried. A classification of these sources in function with their ability to optimize multiphoton processes is proposed, following a protocol found in literature. Starting from these considerations, a suggestion of a possible identikit of the ideal laser source for MPM concludes this topical review. (topical review)

  1. Characterizing lamina propria of human gastric mucosa by multiphoton microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Y C; Yang, H Q; Zhuo, S M [Institute of Laser and Optoelectronics Technology, Fujian Provincial Key Laboratory for Photonics Technology, Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Normal University, Fuzhou 350007 (China); Chen, G; Chen, J X [Department of Pathology, Fujian Provincial Tumor Hospital, Fuzhou, 350014 (China); Yan, J, E-mail: chenjianxin@fjnu.edu.cn, E-mail: ynjun@yahoo.com [Department of Surgery, Fujian Provincial Tumor Hospital, Fuzhou, 350014 (China)

    2011-01-01

    Lamina propria (LP) of gastric mucosa plays an important role in progression of gastric cancer because of the site at where inflammatory reactions occur. Multiphoton imaging has been recently employed for microscopic examination of intact tissue. In this paper, using multiphoton microscopy (MPM) based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG), high resolution multiphoton microscopic images of lamina propria (LP) are obtained in normal human gastric mucosa at excitation wavelength {lambda}{sub ex} = 800 nm. The main source of tissue TPEF originated from the cells of gastric glands, and loose connective tissue, collagen, produced SHG signals. Our results demonstrated that MPM can be effective for characterizing the microstructure of LP in human gastric mucosa. The findings will be helpful for diagnosing and staging early gastric cancer in the clinics.

  2. Characterizing lamina propria of human gastric mucosa by multiphoton microscopy

    Science.gov (United States)

    Liu, Y. C.; Yang, H. Q.; Chen, G.; Zhuo, S. M.; Chen, J. X.; Yan, J.

    2011-01-01

    Lamina propria (LP) of gastric mucosa plays an important role in progression of gastric cancer because of the site at where inflammatory reactions occur. Multiphoton imaging has been recently employed for microscopic examination of intact tissue. In this paper, using multiphoton microscopy (MPM) based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG), high resolution multiphoton microscopic images of lamina propria (LP) are obtained in normal human gastric mucosa at excitation wavelength λex = 800 nm. The main source of tissue TPEF originated from the cells of gastric glands, and loose connective tissue, collagen, produced SHG signals. Our results demonstrated that MPM can be effective for characterizing the microstructure of LP in human gastric mucosa. The findings will be helpful for diagnosing and staging early gastric cancer in the clinics.

  3. The importance of spectroscopy for infrared multiphoton excitation

    International Nuclear Information System (INIS)

    Fuss, W.; Kompa, K.L.

    1980-07-01

    It is substantiated by examples that the infrared spectra of molecules in high vibrational states are similar in width to those of the ground states. Therefore in order to explain collisionless infrared multiphoton excitation, the existence of resonance has to be checked, not only for the first three steps, but for all of them. That is, their (low resolution) spectra should be studied. This review summarizes the spectroscopic mechanisms contributing to multiphoton excitation, which have been suggested to date, including several kinds of rotational compensation and of vibrational level splitting, which cooperate to overcome the anharmonic shift. The spectral quasicontinuum, generated by intensity borrowing, must neither be very broad nor dense, and collisionless vibrational relaxation is only important at very high energies. Knowledge of relatively few spectroscopic detailes helps to understand many details and many differences in multiphoton excitatio. (orig.)

  4. A review of biomedical multiphoton microscopy and its laser sources

    Science.gov (United States)

    Lefort, Claire

    2017-10-01

    Multiphoton microscopy (MPM) has been the subject of major development efforts for about 25 years for imaging biological specimens at micron scale and presented as an elegant alternative to classical fluorescence methods such as confocal microscopy. In this topical review, the main interests and technical requirements of MPM are addressed with a focus on the crucial role of excitation source for optimization of multiphoton processes. Then, an overview of the different sources successfully demonstrated in literature for MPM is presented, and their physical parameters are inventoried. A classification of these sources in function with their ability to optimize multiphoton processes is proposed, following a protocol found in literature. Starting from these considerations, a suggestion of a possible identikit of the ideal laser source for MPM concludes this topical review. Dedicated to Martin.

  5. Intravital and post-mortem CT examinations of cerebral gunshot injuries

    International Nuclear Information System (INIS)

    Schumacher, M.; Oehmichen, M.; Koenig, H.G.; Einighammer, H.; Koeln Univ.; Tuebingen Univ.; Duesseldorf Univ.

    1983-01-01

    The value of CT was assessed in 24 patients who died of cerebral gun-shot injuries and in two patients with more recent injuries in order to reconstruct the mode of injury and for adding forensic information. The post-mortem and intravital appearances are described and are compared with ultrasound rotation compound scans of the isolated brains. CT showed good agreement with pathological findings. Ultrasound produced images with an accuracy between CT and photographs of the brain specimen. Both methods are regarded as valuable additions to the pathological and forensic information concerning gunshot injuries. (orig.) [de

  6. Multiphoton above threshold effects in strong-field fragmentation

    DEFF Research Database (Denmark)

    B Madsen, C; Anis, F; B Madsen, L

    2012-01-01

    We present a study of multiphoton dissociative ionization from molecules. By solving the time-dependent Schrödinger equation for H2+ and projecting the solution onto double continuum scattering states, we observe the correlated electron-nuclear ionization dynamics in detail. We show—for the first...... time—how multiphoton structure prevails as long as one accounts for the energies of all the fragments. Our current work provides a new avenue to analyze strong-field fragmentation that leads to a deeper understanding of the correlated molecular dynamics....

  7. Three-dimensional spatial imaging in multiphoton ionization rate measurements

    International Nuclear Information System (INIS)

    Bredy, Richard; Camp, Howard A.; Nguyen, Hai; Awata, Takaaki; Shan Bing; Chang Zhenghu; DePaola, B.D.

    2004-01-01

    An experiment is described in which an apparatus is used to demonstrate the feasibility of measuring multiphoton photoionization rates in the interaction of short pulsed lasers with atoms or molecules. With this methodology, the ionization rate is measured as a function of the spatial position in the beam-waist region of the laser through the direct three-dimensional spatial imaging of the ionization events. Thus, if the spatial dependence of the laser beam intensity were known, a series of experiments could yield the intensity dependence of multiphoton ionization without the assumptions or errors that are generally inherent in the integration over one or more dimensions in the laser focal volume

  8. Multiphoton ionization for hydrogen plasma diagnostics

    International Nuclear Information System (INIS)

    Bonnie, J.H.M.

    1987-01-01

    In this thesis the processes leading to the formation of negative ions (H - ) in hydrogen discharges are studied. These ions enable efficient production of a beam of fast neutral particles. Such beams are applied in nuclear fusion research. A model has been generally accepted in which H - is formed by means of dissociative attachment (DA) of electrons to vibrationally excited hydrogen molecules [H 2 (υ'')] molecule: when υ'' is low, electron emission is most probable, but when υ'' is high, H - production dominates. A necessary preliminary to the DA process is the presence of sufficient [H 2 (υ'')] molecules with υ'' > 4. By determining the densities of hydrogen molecules in the various vibrational levels as a function of the various discharge parameters (scaling laws), insight can be gained into the extent to which the DA process contributes to H - formation. Since the de-excitation of [H 2 (υ'')] molecules by H atoms is expected to have a large cross section, it is also relevant to determine the scaling laws for atomic hydrogen. This thesis gives an account of the development of an experimental setup for obtaining such measurements, and reports the first results achieved. In view of the anticipated density of the vibrationally excited molecules and the detection limit considered feasible, the diagnostic chosen was resonance-enhanced multiphoton ionization (REMPI). The principle is based on state-selective ionization with REMPI of particles effusing from the discharge chamber through an aperture in the wall. The ions produced in the REMPI-process are then detected. The use of both an electric and a magnetic field makes it possible to distinguish the REMPI ions from those originating elsewhere, such as plasma ions or photodesorption ions. 145 refs.; 25 figs.; 6 tabs

  9. Imaging circulating tumor cells in freely moving awake small animals using a miniaturized intravital microscope.

    Directory of Open Access Journals (Sweden)

    Laura Sarah Sasportas

    Full Text Available Metastasis, the cause for 90% of cancer mortality, is a complex and poorly understood process involving the invasion of circulating tumor cells (CTCs into blood vessels. These cells have potential prognostic value as biomarkers for early metastatic risk. But their rarity and the lack of specificity and sensitivity in measuring them render their interrogation by current techniques very challenging. How and when these cells are circulating in the blood, on their way to potentially give rise to metastasis, is a question that remains largely unanswered. In order to provide an insight into this "black box" using non-invasive imaging, we developed a novel miniature intravital microscopy (mIVM strategy capable of real-time long-term monitoring of CTCs in awake small animals. We established an experimental 4T1-GL mouse model of metastatic breast cancer, in which tumor cells express both fluorescent and bioluminescent reporter genes to enable both single cell and whole body tumor imaging. Using mIVM, we monitored blood vessels of different diameters in awake mice in an experimental model of metastasis. Using an in-house software algorithm we developed, we demonstrated in vivo CTC enumeration and computation of CTC trajectory and speed. These data represent the first reported use we know of for a miniature mountable intravital microscopy setup for in vivo imaging of CTCs in awake animals.

  10. Imaging windows for long-term intravital imaging: General overview and technical insights.

    Science.gov (United States)

    Alieva, Maria; Ritsma, Laila; Giedt, Randy J; Weissleder, Ralph; van Rheenen, Jacco

    2014-01-01

    Intravital microscopy is increasingly used to visualize and quantitate dynamic biological processes at the (sub)cellular level in live animals. By visualizing tissues through imaging windows, individual cells (e.g., cancer, host, or stem cells) can be tracked and studied over a time-span of days to months. Several imaging windows have been developed to access tissues including the brain, superficial fascia, mammary glands, liver, kidney, pancreas, and small intestine among others. Here, we review the development of imaging windows and compare the most commonly used long-term imaging windows for cancer biology: the cranial imaging window, the dorsal skin fold chamber, the mammary imaging window, and the abdominal imaging window. Moreover, we provide technical details, considerations, and trouble-shooting tips on the surgical procedures and microscopy setups for each imaging window and explain different strategies to assure imaging of the same area over multiple imaging sessions. This review aims to be a useful resource for establishing the long-term intravital imaging procedure.

  11. Multicolor fluorescent intravital live microscopy (FILM) for surgical tumor resection in a mouse xenograft model.

    Science.gov (United States)

    Thurber, Greg M; Figueiredo, Jose L; Weissleder, Ralph

    2009-11-30

    Complete surgical resection of neoplasia remains one of the most efficient tumor therapies. However, malignant cell clusters are often left behind during surgery due to the inability to visualize and differentiate them against host tissue. Here we establish the feasibility of multicolor fluorescent intravital live microscopy (FILM) where multiple cellular and/or unique tissue compartments are stained simultaneously and imaged in real time. Theoretical simulations of imaging probe localization were carried out for three agents with specificity for cancer cells, stromal host response, or vascular perfusion. This transport analysis gave insight into the probe pharmacokinetics and tissue distribution, facilitating the experimental design and allowing predictions to be made about the localization of the probes in other animal models and in the clinic. The imaging probes were administered systemically at optimal time points based on the simulations, and the multicolor FILM images obtained in vivo were then compared to conventional pathological sections. Our data show the feasibility of real time in vivo pathology at cellular resolution and molecular specificity with excellent agreement between intravital and traditional in vitro immunohistochemistry. Multicolor FILM is an accurate method for identifying malignant tissue and cells in vivo. The imaging probes distributed in a manner similar to predictions based on transport principles, and these models can be used to design future probes and experiments. FILM can provide critical real time feedback and should be a useful tool for more effective and complete cancer resection.

  12. Multicolor fluorescent intravital live microscopy (FILM for surgical tumor resection in a mouse xenograft model.

    Directory of Open Access Journals (Sweden)

    Greg M Thurber

    2009-11-01

    Full Text Available Complete surgical resection of neoplasia remains one of the most efficient tumor therapies. However, malignant cell clusters are often left behind during surgery due to the inability to visualize and differentiate them against host tissue. Here we establish the feasibility of multicolor fluorescent intravital live microscopy (FILM where multiple cellular and/or unique tissue compartments are stained simultaneously and imaged in real time.Theoretical simulations of imaging probe localization were carried out for three agents with specificity for cancer cells, stromal host response, or vascular perfusion. This transport analysis gave insight into the probe pharmacokinetics and tissue distribution, facilitating the experimental design and allowing predictions to be made about the localization of the probes in other animal models and in the clinic. The imaging probes were administered systemically at optimal time points based on the simulations, and the multicolor FILM images obtained in vivo were then compared to conventional pathological sections. Our data show the feasibility of real time in vivo pathology at cellular resolution and molecular specificity with excellent agreement between intravital and traditional in vitro immunohistochemistry.Multicolor FILM is an accurate method for identifying malignant tissue and cells in vivo. The imaging probes distributed in a manner similar to predictions based on transport principles, and these models can be used to design future probes and experiments. FILM can provide critical real time feedback and should be a useful tool for more effective and complete cancer resection.

  13. Imaging circulating tumor cells in freely moving awake small animals using a miniaturized intravital microscope.

    Science.gov (United States)

    Sasportas, Laura Sarah; Gambhir, Sanjiv Sam

    2014-01-01

    Metastasis, the cause for 90% of cancer mortality, is a complex and poorly understood process involving the invasion of circulating tumor cells (CTCs) into blood vessels. These cells have potential prognostic value as biomarkers for early metastatic risk. But their rarity and the lack of specificity and sensitivity in measuring them render their interrogation by current techniques very challenging. How and when these cells are circulating in the blood, on their way to potentially give rise to metastasis, is a question that remains largely unanswered. In order to provide an insight into this "black box" using non-invasive imaging, we developed a novel miniature intravital microscopy (mIVM) strategy capable of real-time long-term monitoring of CTCs in awake small animals. We established an experimental 4T1-GL mouse model of metastatic breast cancer, in which tumor cells express both fluorescent and bioluminescent reporter genes to enable both single cell and whole body tumor imaging. Using mIVM, we monitored blood vessels of different diameters in awake mice in an experimental model of metastasis. Using an in-house software algorithm we developed, we demonstrated in vivo CTC enumeration and computation of CTC trajectory and speed. These data represent the first reported use we know of for a miniature mountable intravital microscopy setup for in vivo imaging of CTCs in awake animals.

  14. Influence of rotation on multiphoton processes in HF

    International Nuclear Information System (INIS)

    Broeckhove, J.; Feyen, B.; Van Leuven, P.

    1994-01-01

    In this contribution, the authors are concerned with the role of rotational motion in multiphoton processes induced by a laser field of high intensity. The authors use the pseudospectral split operator method for the propagation of the quantum wave-function. The rotation is treated by decomposition of the HF wave-function in its angular momentum components

  15. Multiphoton fluorescence lifetime imaging of chemotherapy distribution in solid tumors

    Science.gov (United States)

    Carlson, Marjorie; Watson, Adrienne L.; Anderson, Leah; Largaespada, David A.; Provenzano, Paolo P.

    2017-11-01

    Doxorubicin is a commonly used chemotherapeutic employed to treat multiple human cancers, including numerous sarcomas and carcinomas. Furthermore, doxorubicin possesses strong fluorescent properties that make it an ideal reagent for modeling drug delivery by examining its distribution in cells and tissues. However, while doxorubicin fluorescence and lifetime have been imaged in live tissue, its behavior in archival samples that frequently result from drug and treatment studies in human and animal patients, and murine models of human cancer, has to date been largely unexplored. Here, we demonstrate imaging of doxorubicin intensity and lifetimes in archival formalin-fixed paraffin-embedded sections from mouse models of human cancer with multiphoton excitation and multiphoton fluorescence lifetime imaging microscopy (FLIM). Multiphoton excitation imaging reveals robust doxorubicin emission in tissue sections and captures spatial heterogeneity in cells and tissues. However, quantifying the amount of doxorubicin signal in distinct cell compartments, particularly the nucleus, often remains challenging due to strong signals in multiple compartments. The addition of FLIM analysis to display the spatial distribution of excited state lifetimes clearly distinguishes between signals in distinct compartments such as the cell nuclei versus cytoplasm and allows for quantification of doxorubicin signal in each compartment. Furthermore, we observed a shift in lifetime values in the nuclei of transformed cells versus nontransformed cells, suggesting a possible diagnostic role for doxorubicin lifetime imaging to distinguish normal versus transformed cells. Thus, data here demonstrate that multiphoton FLIM is a highly sensitive platform for imaging doxorubicin distribution in normal and diseased archival tissues.

  16. Multi-photon microscope driven by novel green laser pump

    DEFF Research Database (Denmark)

    Marti, Dominik; Djurhuus, Martin; Jensen, Ole Bjarlin

    2016-01-01

    Multi-photon microscopy is extensively used in research due to its superior possibilities when compared to other microscopy modalities. The technique also has the possibility to advance diagnostics in clinical applications, due to its capabilities complementing existing technology in a multimodal...

  17. Line broadening in multiphoton processes with a resonant intermediate transition

    International Nuclear Information System (INIS)

    Wang, C.C.; James, J.V.; Xia, J.

    1983-01-01

    The linewidth of the excitation spectrum for multiphoton ionization is found to be broadened much more severely than the cascade fluorescence originating from the resonant intermediate level. These results are due to the mutual effects of the ionizing and resonating transitions, which are not properly accounted for in perturbative treatments

  18. Automated Identification and Localization of Hematopoietic Stem Cells in 3D Intravital Microscopy Data

    Directory of Open Access Journals (Sweden)

    Reema A. Khorshed

    2015-07-01

    Full Text Available Measuring three-dimensional (3D localization of hematopoietic stem cells (HSCs within the bone marrow microenvironment using intravital microscopy is a rapidly expanding research theme. This approach holds the key to understanding the detail of HSC-niche interactions, which are critical for appropriate stem cell function. Due to the complex tissue architecture of the bone marrow and to the progressive introduction of scattering and signal loss at increasing imaging depths, there is no ready-made software to handle efficient segmentation and unbiased analysis of the data. To address this, we developed an automated image analysis tool that simplifies and standardizes the biological interpretation of 3D HSC microenvironment images. The algorithm identifies HSCs and measures their localization relative to surrounding osteoblast cells and bone collagen. We demonstrate here the effectiveness, consistency, and accuracy of the proposed approach compared to current manual analysis and its wider applicability to analyze other 3D bone marrow components.

  19. Integrated intravital microscopy and mathematical modeling to optimize nanotherapeutics delivery to tumors

    Directory of Open Access Journals (Sweden)

    Anne L. van de Ven

    2012-03-01

    Full Text Available Inefficient vascularization hinders the optimal transport of cell nutrients, oxygen, and drugs to cancer cells in solid tumors. Gradients of these substances maintain a heterogeneous cell-scale microenvironment through which drugs and their carriers must travel, significantly limiting optimal drug exposure. In this study, we integrate intravital microscopy with a mathematical model of cancer to evaluate the behavior of nanoparticle-based drug delivery systems designed to circumvent biophysical barriers. We simulate the effect of doxorubicin delivered via porous 1000 x 400 nm plateloid silicon particles to a solid tumor characterized by a realistic vasculature, and vary the parameters to determine how much drug per particle and how many particles need to be released within the vasculature in order to achieve remission of the tumor. We envision that this work will contribute to the development of quantitative measures of nanoparticle design and drug loading in order to optimize cancer treatment via nanotherapeutics.

  20. Combined application of dynamic light scattering imaging and fluorescence intravital microscopy in vascular biology

    International Nuclear Information System (INIS)

    Kalchenko, V; Harmelin, A; Ziv, K; Addadi, Y; Madar-Balakirski, N; Neeman, M; Meglinski, I

    2010-01-01

    The dynamic light scattering imaging (DLSI) system combined with the conventional fluorescence intravital microscope (FIM) has been applied for the examination of blood and lymph vessels in the mouse ear in vivo. While the CCD camera can be shared by both techniques the combined application of DLSI and FIM allows rapid switching between the modalities. In current study temporal speckles fluctuations are used for rendering blood vessels structure and monitoring blood perfusion with the higher spatial resolution, whereas FIM provides the images of lymphatic vessels. The results clearly demonstrate that combined application of DLSI and FIM approaches provides synchronic in vivo images of blood and lymph vessels with higher contrast and specificity. The use of this new dual-modal diagnostic system is particularly important and has a great potential to significantly expand the capabilities of vascular diagnostics providing synchronic in vivo images of blood and lymph vessels

  1. Intravital Microscopy Reveals Differences in the Kinetics of Endocytic Pathways between Cell Cultures and Live Animals

    Directory of Open Access Journals (Sweden)

    Roberto Weigert

    2012-11-01

    Full Text Available Intravital microscopy has enabled imaging of the dynamics of subcellular structures in live animals, thus opening the door to investigating membrane trafficking under physiological conditions. Here, we sought to determine whether the architecture and the environment of a fully developed tissue influences the dynamics of endocytic processes. To this aim, we imaged endocytosis in the stromal cells of rat salivary glands both in situ and after they were isolated and cultured on a solid surface. We found that the internalization of transferrin and dextran, two molecules that traffic via distinct mechanisms, is substantially altered in cultured cells, supporting the idea that the three dimensional organization of the tissue and the cues generated by the surrounding environment strongly affect membrane trafficking events.

  2. Automated filtering of intrinsic movement artifacts during two-photon intravital microscopy.

    Directory of Open Access Journals (Sweden)

    Denis Soulet

    Full Text Available In vivo imaging using two-photon microscopy is an essential tool to explore the dynamic of physiological events deep within biological tissues for short or extended periods of time. The new capabilities offered by this technology (e.g. high tissue penetrance, low toxicity have opened a whole new era of investigations in modern biomedical research. However, the potential of using this promising technique in tissues of living animals is greatly limited by the intrinsic irregular movements that are caused by cardiac and respiratory cycles and muscular and vascular tone. Here, we show real-time imaging of the brain, spinal cord, sciatic nerve and myenteric plexus of living mice using a new automated program, named Intravital_Microscopy_Toolbox, that removes frames corrupted with motion artifacts from time-lapse videos. Our approach involves generating a dissimilarity score against precalculated reference frames in a specific reference channel, thus allowing the gating of distorted, out-of-focus or translated frames. Since the algorithm detects the uneven peaks of image distortion caused by irregular animal movements, the macro allows a fast and efficient filtering of the image sequence. In addition, extra features have been implemented in the macro, such as XY registration, channel subtraction, extended field of view with maximum intensity projection, noise reduction with average intensity projections, and automated timestamp and scale bar overlay. Thus, the Intravital_Microscopy_Toolbox macro for ImageJ provides convenient tools for biologists who are performing in vivo two-photon imaging in tissues prone to motion artifacts.

  3. Percutaneous window chamber method for chronic intravital microscopy of sensor-tissue interactions.

    Science.gov (United States)

    Koschwanez, Heidi E; Klitzman, Bruce; Reichert, W Monty

    2008-11-01

    A dorsal, two-sided skin-fold window chamber model was employed previously by Gough in glucose sensor research to characterize poorly understood physiological factors affecting sensor performance. We have extended this work by developing a percutaneous one-sided window chamber model for the rodent dorsum that offers both a larger subcutaneous area and a less restrictive tissue space than previous animal models. A surgical procedure for implanting a sensor into the subcutis beneath an acrylic window (15 mm diameter) is presented. Methods to quantify changes in the microvascular network and red blood cell perfusion around the sensors using noninvasive intravital microscopy and laser Doppler flowmetry are described. The feasibility of combining interstitial glucose monitoring from an implanted sensor with intravital fluorescence microscopy was explored using a bolus injection of fluorescein and dextrose to observe real-time mass transport of a small molecule at the sensor-tissue interface. The percutaneous window chamber provides an excellent model for assessing the influence of different sensor modifications, such as surface morphologies, on neovascularization using real-time monitoring of the microvascular network and tissue perfusion. However, the tissue response to an implanted sensor was variable, and some sensors migrated entirely out of the field of view and could not be observed adequately. A percutaneous optical window provides direct, real-time images of the development and dynamics of microvascular networks, microvessel patency, and fibrotic encapsulation at the tissue-sensor interface. Additionally, observing microvessels following combined bolus injections of a fluorescent dye and glucose in the local sensor environment demonstrated a valuable technique to visualize mass transport at the sensor surface.

  4. Applications of multiphoton microscopy in the field of colorectal cancer

    Science.gov (United States)

    Wang, Shu; Li, Lianhuang; Zhu, Xiaoqin; Zheng, Liqin; Zhuo, Shuangmu; Chen, Jianxin

    2018-06-01

    Multiphoton microscopy (MPM) is a powerful tool for visualizing cellular and subcellular details within living tissue by its unique advantages of being label-free, its intrinsic optical sectioning ability, near-infrared excitation for deep penetration depth into tissue, reduced photobleaching and phototoxicity in the out-of-focus regions, and being capable of providing quantitative information. In this review, we focus on applications of MPM in the field of colorectal cancer, including monitoring cancer progression, detecting tumor metastasis and microenvironment, evaluating the cancer therapy response, and visualizing and ablating pre-invasive cancer cells. We also present one of the major challenges and the future research direction to exploit a colorectal multiphoton endoscope.

  5. Characteristics of subgingival calculus detection by multiphoton fluorescence microscopy

    Science.gov (United States)

    Tung, Oi-Hong; Lee, Shyh-Yuan; Lai, Yu-Lin; Chen, How-Foo

    2011-06-01

    Subgingival calculus has been recognized as a major cause of periodontitis, which is one of the main chronic infectious diseases of oral cavities and a principal cause of tooth loss in humans. Bacteria deposited in subgingival calculus or plaque cause gingival inflammation, function deterioration, and then periodontitis. However, subgingival calculus within the periodontal pocket is a complicated and potentially delicate structure to be detected with current dental armamentaria, namely dental x-rays and dental probes. Consequently, complete removal of subgingival calculus remains a challenge to periodontal therapies. In this study, the detection of subgingival calculus employing a multiphoton autofluorescence imaging method was characterized in comparison with a one-photon confocal fluorescence imaging technique. Feasibility of such a system was studied based on fluorescence response of gingiva, healthy teeth, and calculus with and without gingiva covered. The multiphoton fluorescence technology perceived the tissue-covered subgingival calculus that cannot be observed by the one-photon confocal fluorescence method.

  6. High-order multiphoton ionization photoelectron spectroscopy of NO

    International Nuclear Information System (INIS)

    Carman, H.S. Jr.; Compton, R.N.

    1987-01-01

    Photoelectron energy angular distributions of NO following three different high-order multiphoton ionization (MPI) schemes have been measured. The 3 + 3 resonantly enhanced multiphoton ionization (REMPI) via the A 2 Σ + (v=O) level yielded a distribution of electron energies corresponding to all accessible vibrational levels (v + =O-6) of the nascent ion. Angular distributions of electrons corresponding to v + =O and v + =3 were significantly different. The 3 + 2 REMPI via the A 2 Σ + (v=1) level produced only one low-energy electron peak (v + =1). Nonresonant MPI at 532 nm yielded a distribution of electron energies corresponding to both four- and five-photon ionization. Prominent peaks in the five-photon photoelectron spectrum (PES) suggest contributions from near-resonant states at the three-photon level. 4 refs., 3 figs

  7. Scanless multitarget-matching multiphoton excitation fluorescence microscopy

    Directory of Open Access Journals (Sweden)

    Junpeng Qiu

    2018-03-01

    Full Text Available Using the combination of a reflective blazed grating and a reflective phase-only diffractive spatial light modulator (SLM, scanless multitarget-matching multiphoton excitation fluorescence microscopy (SMTM-MPM was achieved. The SLM shaped an incoming mode-locked, near-infrared Ti:sapphire laser beam into an excitation pattern with addressable shapes and sizes that matched the samples of interest in the field of view. Temporal and spatial focusing were simultaneously realized by combining an objective lens and a blazed grating. The fluorescence signal from illuminated areas was recorded by a two-dimensional sCMOS camera. Compared with a conventional temporal focusing multiphoton microscope, our microscope achieved effective use of the laser power and decreased photodamage with higher axial resolution.

  8. Rapid in vivo vertical tissue sectioning by multiphoton tomography

    Science.gov (United States)

    Batista, Ana; Breunig, Hans Georg; König, Karsten

    2018-02-01

    A conventional tool in the pathological field is histology which involves the analysis of thin sections of tissue in which specific cellular structures are stained with different dyes. The process to obtain these stained tissue sections is time consuming and invasive as it requires tissue removal, fixation, sectioning, and staining. Moreover, imaging of live tissue is not possible. We demonstrate that multiphoton tomography can provide within seconds, non-invasive, label-free, vertical images of live tissue which are in quality similar to conventional light micrographs of histologic stained specimen. In contrast to conventional setups based on laser scanning which image horizontally sections, the vertical in vivo images are directly recorded by combined line scanning and timed adjustments of the height of the focusing optics. In addition, multiphoton tomography provides autofluorescence lifetimes which can be used to determine the metabolic states of cells.

  9. Multiphoton absorption coefficients in solids: an universal curve

    International Nuclear Information System (INIS)

    Brandi, H.S.; Araujo, C.B. de

    1983-04-01

    An universal curve for the frequency dependence of the multiphoton absorption coefficient is proposed based on a 'non-perturbative' approach. Specific applications have been made to obtain two, three, four and five photons absorption coefficient in different materials. Properly scaling of the two photon absorption coefficient and the use of the universal curve yields results for the higher order absorption coefficients in good agreement with the experimental data. (Author) [pt

  10. Femtosecond Light Source for Phase-Controlled Multiphoton Ionization

    International Nuclear Information System (INIS)

    Sokolov, A. V.; Walker, D. R.; Yavuz, D. D.; Yin, G. Y.; Harris, S. E.

    2001-01-01

    We describe a femtosecond Raman light source with more than an octave of optical bandwidth. We use this source to demonstrate phase control of multiphoton ionization under conditions where ionization requires eleven photons of the lowest frequency of the spectrum or five photons of the highest frequency. The nonlinearity of the photoionization process allows us to characterize the light source. Experiment-to-theory comparison implies generation of a near single-cycle waveform

  11. Multi-photon UV photolysis of gaseous polycyclic aromatic hydrocarbons: Extinction spectra and dynamics

    Energy Technology Data Exchange (ETDEWEB)

    Walsh, A. J.; Gash, E. W.; Mansfield, M. W. D. [Physics Department, University College Cork, Cork (Ireland); Ruth, A. A. [Physics Department, University College Cork, Cork (Ireland); Environmental Research Institute, University College Cork, Cork (Ireland)

    2013-08-07

    The extinction spectra of static naphthalene and static biphenylene vapor, each buffered with a noble gas at room temperature, were measured as a function of time in the region between 390 and 850 nm after UV multi-photon laser photolysis at 308 nm. Employing incoherent broadband cavity enhanced absorption spectroscopy (IBBCEAS), the spectra were found to be unstructured with a general lack of isolated features suggesting that the extinction was not solely based on absorption but was in fact dominated by scattering from particles formed in the photolysis of the respective polycyclic aromatic hydrocarbon. Following UV multi-photon photolysis, the extinction dynamics of the static (unstirred) closed gas-phase system exhibits extraordinary quasi-periodic and complex oscillations with periods ranging from seconds to many minutes, persisting for up to several hours. Depending on buffer gas type and pressure, several types of dynamical responses could be generated (classified as types I, II, and III). They were studied as a function of temperature and chamber volume for different experimental conditions and possible explanations for the oscillations are discussed. A conclusive model for the observed phenomena has not been established. However, a number of key hypotheses have made based on the measurements in this publication: (a) Following the multi-photon UV photolysis of naphthalene (or biphenylene), particles are formed on a timescale not observable using IBBCEAS. (b) The observed temporal behavior cannot be described on basis of a chemical reaction scheme alone. (c) The pressure dependence of the system's responses is due to transport phenomena of particles in the chamber. (d) The size distribution and the refractive indices of particles are time dependent and evolve on a timescale of minutes to hours. The rate of particle coagulation, involving coalescent growth and particle agglomeration, affects the observed oscillations. (e) The walls of the chamber act as a

  12. Visualizing radiofrequency-skin interaction using multiphoton microscopy in vivo.

    Science.gov (United States)

    Tsai, Tsung-Hua; Lin, Sung-Jan; Lee, Woan-Ruoh; Wang, Chun-Chin; Hsu, Chih-Ting; Chu, Thomas; Dong, Chen-Yuan

    2012-02-01

    Redundant skin laxity is a major feature of aging. Recently, radiofrequency has been introduced for nonablative tissue tightening by volumetric heating of the deep dermis. Despite the wide range of application based on this therapy, the effect of this technique on tissue and the subsequent tissue remodeling have not been investigated in detail. Our objective is to evaluate the potential of non-linear optics, including multiphoton autofluorescence and second harmonic generation (SHG) microscopy, as a non-invasive imaging modality for the real-time study of radiofrequency-tissue interaction. Electro-optical synergy device (ELOS) was used as the radiofrequency source in this study. The back skin of nude mouse was irradiated with radiofrequency at different passes. We evaluated the effect on skin immediately and 1 month after treatment with multiphoton microscopy. Corresponding histology was performed for comparison. We found that SHG is negatively correlated to radiofrequency passes, which means that collagen structural disruption happens immediately after thermal damage. After 1 month of collagen remodeling, SHG signals increased above baseline, indicating that collagen regeneration has occurred. Our findings may explain mechanism of nonablative skin tightening and were supported by histological examinations. Our work showed that monitoring the dermal heating status of RF and following up the detailed process of tissue reaction can be imaged and quantified with multiphoton microscopy non-invasively in vivo. Copyright © 2011. Published by Elsevier Ireland Ltd.

  13. Uptake of Retinoic Acid-Modified PMMA Nanoparticles in LX-2 and Liver Tissue by Raman Imaging and Intravital Microscopy.

    Science.gov (United States)

    Yildirim, Turgay; Matthäus, Christian; Press, Adrian T; Schubert, Stephanie; Bauer, Michael; Popp, Jürgen; Schubert, Ulrich S

    2017-10-01

    A primary amino-functionalized methyl methacrylate-based statistical copolymer is covalently coupled with retinoic acid (RA) and a fluorescent dye (DY590) in order to investigate the feasibility of the RA containing polymeric nanoparticles for Raman imaging studies and to study the possible selectivity of RA for hepatic stellate cells via intravital microscopy. Cationic nanoparticles are prepared by utilizing the nanoprecipitation method using modified polymers. Raman studies show that RA functional nanoparticles can be detectable in all tested cells without any need of additional label. Moreover, intravital microscopy indicates that DY590 is eliminated through the hepatobiliary route but not if used as covalently attached tracing molecule for nanoparticles. However, it is a suitable probe for sensitive detection of polymeric nanoparticles. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Intravital imaging of the immune responses during liver-stage malaria infection: An improved approach for fixing the liver.

    Science.gov (United States)

    Akbari, Masoud; Kimura, Kazumi; Houts, James T; Yui, Katsuyuki

    2016-10-01

    The host-parasite relationship is one of the main themes of modern parasitology. Recent revolutions in science, including the development of various fluorescent proteins/probes and two-photon microscopy, have made it possible to directly visualize and study the mechanisms underlying the interaction between the host and pathogen. Here, we describe our method of preparing and setting-up the liver for our experimental approach of using intravital imaging to examine the interaction between Plasmodium berghei ANKA and antigen-specific CD8 + T cells during the liver-stage of the infection in four dimensions. Since the liver is positioned near the diaphragm, neutralization of respiratory movements is critical during the imaging process. In addition, blood circulation and temperature can be affected by the surgical exposure due to the anatomy and tissue structure of the liver. To control respiration, we recommend anesthesia with isoflurane inhalation at 1% during the surgery. In addition, our protocol introduces a cushion of gauze around the liver to avoid external pressure on the liver during intravital imaging using an inverted microscope, which makes it possible to image the liver tissue for long periods with minimal reduction in the blood circulation and with minimal displacement and tissue damage. The key point of this method is to reduce respiratory movements and external pressure on the liver tissue during intravital imaging. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  15. 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.

  16. In vivo imaging of leukocyte recruitment to glomeruli in mice using intravital microscopy.

    Science.gov (United States)

    Kitching, A Richard; Kuligowski, Michael P; Hickey, Michael J

    2009-01-01

    Leukocytes mediate some forms of glomerulonephritis, particularly severe proliferative and crescentic forms. The renal glomerulus is one of the few sites within the microvasculature in which leukocyte recruitment occurs in capillaries. However, due to the difficulty of directly visualising the glomerulus, the mechanisms of leukocyte recruitment to glomerular capillaries are poorly understood. To overcome this, a murine kidney can be rendered hydronephrotic, by ligating one ureter, and allowing the mouse to rest for 12 weeks. This allows the visualisation of the glomerular microvasculature during inflammatory responses. In inflammation, in this example induced by anti-glomerular basement membrane (GBM) antibody, leukocytes can be observed undergoing adhesion in glomerular capillaries using intravital microscopy. Leukocyte adhesion can be quantitated using this approach. An observation protocol involving few, limited periods of epifluorescence avoids phototoxicity-induced leukocyte recruitment. The process of hydronephrosis does not alter the ability of anti-GBM-antibody to induce a glomerular inflammatory response. This approach allows detailed investigation of the mechanisms of leukocyte recruitment within glomeruli.

  17. Intravital imaging reveals transient changes in pigment production and Brn2 expression during metastatic melanoma dissemination.

    Science.gov (United States)

    Pinner, Sophie; Jordan, Peter; Sharrock, Kirsty; Bazley, Laura; Collinson, Lucy; Marais, Richard; Bonvin, Elise; Goding, Colin; Sahai, Erik

    2009-10-15

    How melanoma acquire a metastatic phenotype is a key issue. One possible mechanism is that metastasis is driven by microenvironment-induced switching between noninvasive and invasive states. However, whether switching is a reversible or hierarchical process is not known and is difficult to assess by comparison of primary and metastatic tumors. We address this issue in a model of melanoma metastasis using a novel intravital imaging method for melanosomes combined with a reporter construct in which the Brn-2 promoter drives green fluorescent protein (GFP) expression. A subpopulation of cells containing little or no pigment and high levels of Brn2::GFP expression are motile in the primary tumor and enter the vasculature. Significantly, the less differentiated state of motile and intravasated cells is not maintained at secondary sites, implying switching between states as melanoma cells metastasize. We show that melanoma cells can switch in both directions between high- and low-pigment states. However, switching from Brn2::GFP high to low was greatly favored over the reverse direction. Microarray analysis of high- and low-pigment populations revealed that transforming growth factor (TGF)beta2 was up-regulated in the poorly pigmented cells. Furthermore, TGFbeta signaling induced hypopigmentation and increased cell motility. Thus, a subset of less differentiated cells exits the primary tumor but subsequently give rise to metastases that include a range of more differentiated and pigment-producing cells. These data show reversible phenotype switching during melanoma metastasis.

  18. Non-perturbative methods applied to multiphoton ionization

    International Nuclear Information System (INIS)

    Brandi, H.S.; Davidovich, L.; Zagury, N.

    1982-09-01

    The use of non-perturbative methods in the treatment of atomic ionization is discussed. Particular attention is given to schemes of the type proposed by Keldysh where multiphoton ionization and tunnel auto-ionization occur for high intensity fields. These methods are shown to correspond to a certain type of expansion of the T-matrix in the intra-atomic potential; in this manner a criterium concerning the range of application of these non-perturbative schemes is suggested. A brief comparison between the ionization rate of atoms in the presence of linearly and circularly polarized light is presented. (Author) [pt

  19. Coherent enhancement of resonance-mediated multiphoton absorption

    International Nuclear Information System (INIS)

    Zhang, Shian; Zhang, Hui; Jia, Tianqing; Wang, Zugeng; Sun, Zhenrong

    2010-01-01

    In this paper, we theoretically investigate the coherent enhancement of resonance-mediated (2+2) four-photon absorption. It is found that by shaping the spectral phase with a π phase step, the resonance-mediated (2+2) four-photon transition probability can be enhanced. Furthermore, the coherent enhancement dependences on the detuning between the two two-photon absorptions, laser spectral bandwidth and laser centre frequency are explicitly discussed and analysed. We believe these theoretical results may play an important role in enhancing more complex resonance-mediated multiphoton absorption processes.

  20. Multi-photon microscope driven by novel green laser pump

    Science.gov (United States)

    Marti, Dominik; Djurhuus, Martin; Jensen, Ole Bjarlin; Andersen, Peter E.

    2016-03-01

    Multi-photon microscopy is extensively used in research due to its superior possibilities when compared to other microscopy modalities. The technique also has the possibility to advance diagnostics in clinical applications, due to its capabilities complementing existing technology in a multimodal system. However, translation is hindered due to the high cost, high training demand and large footprint of a standard setup. We show in this article that minification of the setup, while also reducing cost and complexity, is indeed possible without compromising on image quality, by using a novel diode laser replacing the commonly used conventional solid state laser as the pump for the femtosecond system driving the imaging.

  1. Microstructure imaging of human rectal mucosa using multiphoton microscopy

    Science.gov (United States)

    Liu, N. R.; Chen, G.; Chen, J. X.; Yan, J.; Zhuo, S. M.; Zheng, L. Q.; Jiang, X. S.

    2011-01-01

    Multiphoton microscopy (MPM) has high resolution and sensitivity. In this study, MPM was used to image microstructure of human rectal mucosa. The morphology and distribution of the main components in mucosa layer, absorptive cells and goblet cells in the epithelium, abundant intestinal glands in the lamina propria and smooth muscle fibers in the muscularis mucosa were clearly monitored. The variations of these components were tightly relevant to the pathology in gastrointestine system, especially early rectal cancer. The obtained images will be helpful for the diagnosis of early colorectal cancer.

  2. The study of multiphoton ionization processes in hydrogen atoms

    International Nuclear Information System (INIS)

    Mohammad, M.A.

    1981-01-01

    In this thesis we investigate theoretically the multiphoton ionization of hydrogen atoms based on perturbation theory.The main problem in the numorical evaluation is the appearance of infinite summation over the matrix element and energy denominators of the intermediate state in the formula for ionization cross section.Our numerical result is in excellent agreement with other workers.In the last part of the thesis we have again calculated the two photon ionization of hydrogen atoms using momentum translation approximation of Reiss.The method in general is in fair agreement with other calculations but dose not show the resonance behaviour.(2 tabs., 1 fig., 45 refs.)

  3. Multiphoton photodegradation of indocyanine green: Solvent protolysis effect

    Energy Technology Data Exchange (ETDEWEB)

    Fuyuki, Masanori, E-mail: mn.fuyuki@kio.ac.jp

    2016-02-15

    The multiphoton photodegradation mechanism of indocyanine green (ICG) was investigated by using femtosecond near-infrared (NIR) pump and probe pulses. In the pump fluence region from 2 mJ/cm{sup 2} to 4 mJ/cm{sup 2}, the photodegradation rate was higher in acetic acid than in ethanol, and the rate was proportional to pump fluence to the 2.3th power in acetic acid and the 3.9th in ethanol. Considering that the degree of auto-protolysis of acetic acid is much higher than that of ethanol, the experimental results indicate that self-ionized solvent molecules played an essential role in the degradation of ICG molecules excited by NIR multiphoton process. - Highlights: • Photodegradation of ICG by femtosecond near-infrared pulses. • Photodegradation rate of ICG was higher in acetic acid than in ethanol. • Photodegradation rate was proportional to pump fluence to 2.3th power in acetic acid. • Photodegradation rate was proportional to pump fluence to 3.9th power in ethanol. • Self-ionized solvent molecules promoted ICG photodegradation in acetic acid.

  4. Experimental REMPI [Resonance Enhanced Multiphoton Ionization] studies of small molecules

    International Nuclear Information System (INIS)

    Dehmer, J.L.; Dehmer, P.M.; Pratt, S.T.; O'Halloran, M.A.; Tomkins, F.S.

    1986-01-01

    Resonance Enhanced Multiphoton Ionization (REMPI) utilizes tunable dye lasers to ionize an atom or molecule by first preparing an excited state by multiphoton absorption and then ionizing that state before it can decay. This process is highly selective with respect to both the initial and resonant intermediate states of the target, and it can be extremely sensitive. In addition, the products of the REMPI process can be detected as needed by analyzing the resulting electrons, ions, fluorescence, or by additional REMPI. This points to a number of exciting opportunities for both basic and applied science. On the applied side, REMPI has great potential as an ultrasensitive, highly selective detector for trace, reactive, or transient species. On the basic side, REMPI affords an unprecedented means of exploring excited state physics and chemistry at the quantum-state-specific level. We shall give an overview together with examples of current studies of excited molecular states to illustrate the principles of and prospects for REMPI. 27 refs., 3 figs

  5. Intravital Imaging Reveals Angiotensin II–Induced Transcytosis of Albumin by Podocytes

    Science.gov (United States)

    Schießl, Ina Maria; Hammer, Anna; Kattler, Veronika; Gess, Bernhard; Theilig, Franziska; Witzgall, Ralph

    2016-01-01

    Albuminuria is a hallmark of kidney disease of various etiologies and usually caused by deterioration of glomerular filtration barrier integrity. We recently showed that angiotensin II (Ang II) acutely increases albumin filtration in the healthy kidney. Here, we used intravital microscopy to assess the effects of Ang II on podocyte function in rats. Acute infusion of 30, 60, or 80 ng/kg per minute Ang II enhanced the endocytosis of albumin by activation of the type 1 Ang II receptor and resulted in an average (±SEM) of 3.7±2.2, 72.3±18.6 (Palbumin-containing vesicles per glomerulus, respectively, compared with none at baseline or 10 ng/kg per minute Ang II. Immunostaining of Ang II–infused kidneys confirmed the presence of albumin-containing vesicles, which colocalized with megalin, in podocin-positive cells. Furthermore, podocyte endocytosis of albumin was markedly reduced in the presence of gentamicin, a competitive inhibitor of megalin-dependent endocytosis. Ang II infusion increased the concentration of albumin in the subpodocyte space, a potential source for endocytic protein uptake, and gentamicin further increased this concentration. Some endocytic vesicles were acidified and colocalized with LysoTracker. Most vesicles migrated from the capillary to the apical aspect of the podocyte and were eventually released into the urinary space. This transcytosis accounted for approximately 10% of total albumin filtration. In summary, the transcellular transport of proteins across the podocyte constitutes a new pathway of glomerular protein filtration. Ang II enhances the endocytosis and transcytosis of plasma albumin by podocytes, which may eventually impair podocyte function. PMID:26116357

  6. Intravital Imaging Reveals Angiotensin II-Induced Transcytosis of Albumin by Podocytes.

    Science.gov (United States)

    Schießl, Ina Maria; Hammer, Anna; Kattler, Veronika; Gess, Bernhard; Theilig, Franziska; Witzgall, Ralph; Castrop, Hayo

    2016-03-01

    Albuminuria is a hallmark of kidney disease of various etiologies and usually caused by deterioration of glomerular filtration barrier integrity. We recently showed that angiotensin II (Ang II) acutely increases albumin filtration in the healthy kidney. Here, we used intravital microscopy to assess the effects of Ang II on podocyte function in rats. Acute infusion of 30, 60, or 80 ng/kg per minute Ang II enhanced the endocytosis of albumin by activation of the type 1 Ang II receptor and resulted in an average (±SEM) of 3.7±2.2, 72.3±18.6 (Palbumin-containing vesicles per glomerulus, respectively, compared with none at baseline or 10 ng/kg per minute Ang II. Immunostaining of Ang II-infused kidneys confirmed the presence of albumin-containing vesicles, which colocalized with megalin, in podocin-positive cells. Furthermore, podocyte endocytosis of albumin was markedly reduced in the presence of gentamicin, a competitive inhibitor of megalin-dependent endocytosis. Ang II infusion increased the concentration of albumin in the subpodocyte space, a potential source for endocytic protein uptake, and gentamicin further increased this concentration. Some endocytic vesicles were acidified and colocalized with LysoTracker. Most vesicles migrated from the capillary to the apical aspect of the podocyte and were eventually released into the urinary space. This transcytosis accounted for approximately 10% of total albumin filtration. In summary, the transcellular transport of proteins across the podocyte constitutes a new pathway of glomerular protein filtration. Ang II enhances the endocytosis and transcytosis of plasma albumin by podocytes, which may eventually impair podocyte function. Copyright © 2016 by the American Society of Nephrology.

  7. Multiphoton imaging with a novel compact diode-pumped Ti:sapphire oscillator

    DEFF Research Database (Denmark)

    König, Karsten; Andersen, Peter E.; Le, Tuan

    2015-01-01

    Multiphoton laser scanning microscopy commonly relies on bulky and expensive femtosecond lasers. We integrated a novel minimal-footprint Ti:sapphire oscillator, pumped by a frequency-doubled distributed Bragg reflector tapered diode laser, into a clinical multiphoton tomograph and evaluated its...... imaging capability using different biological samples, i.e. cell monolayers, corneal tissue, and human skin. With the novel laser, the realization of very compact Ti:sapphire-based systems for high-quality multiphoton imaging at a significantly size and weight compared to current systems will become...

  8. Effects of laser radiation parameters of the infrared multiphoton dissociation of protonated trichloroethylene

    International Nuclear Information System (INIS)

    Ungureanu, C.; Almasan, V.

    1994-01-01

    The favorable properties of the infrared multiphoton absorption and dissociation of trichloroethylene-H, (C 2 HCl 3 ), by TEA-CO 2 laser radiation and rapid isotopic exchange between this molecule and water, indicate that it can be a promising further candidate for the final enrichment of heavy water (> 98% D 2 O), by laser method. We present the results obtained in the isotopic selectivity of multiphoton absorption measurements and in the study of the pulse energy and frequency laser radiation influence on the infrared multiphoton dissociation of C 2 HCl 3 in isotopic mixture with C 2 DCl 3 . (Author)

  9. Multiphoton ionization as a probe of molecular photofragmentation: statistical and dynamical energy partitioning in the multiphoton dissociation of nitromethane

    International Nuclear Information System (INIS)

    Rockney, B.H.

    1982-01-01

    Multiphoton ionization (MPI) appears in its first use as a probe of laser-induced photofragmentation. Specifically, MPI here reveals the internal and translational energy content of the nascent fragments from the infrared multiphoton dissociation (MPD) of nitromethane (CH 3 NO 2 ). The apparatus for this work consists of a pulsed supersonic molecular beam crossed by two pulsed and focused lasers - a CO 2 laser to induce collision-free unimolecular dissociation of CH 3 NO 2 , and a tunable dye laser following immediately to ionize selectively one of the pair of dissociation fragments for detection by a mass spectrometer and particle multiplier. A computer simulation of each fragment's MPI spectrum, a series of four photon resonances to members of the npsigma/sub u/ Rydberg state of NO 2 and three photon resonances to two vibrational members of the #betta# 1 Rydberg state of CH 3 , aids in determining the fragment's internal energy content. The dye laser is delayed and its focus is traced through a small quarter circle centered at the focus of the CO 2 laser. The flight times of the fragments from the point of dissociation and their laboratory scattering angular distributions at fixed ionizing laser wavelength provide their center of mass recoil velocity distributions. The energy deposited in the fragments evidences a striking mixture of statistical and dynamical energy partitioning. The statistical RRKM theory of unimolecular decomposition accurately predicts the amount of internal energy found in the fragments

  10. Intravital imaging of donor allogeneic effector and regulatory T cells with host dendritic cells during GVHD.

    Science.gov (United States)

    Lin, Kaifeng Lisa; Fulton, LeShara M; Berginski, Matthew; West, Michelle L; Taylor, Nicholas A; Moran, Timothy P; Coghill, James M; Blazar, Bruce R; Bear, James E; Serody, Jonathan S

    2014-03-06

    Graft-versus-host disease (GVHD) is a systemic inflammatory response due to the recognition of major histocompatibility complex disparity between donor and recipient after hematopoietic stem cell transplantation (HSCT). T-cell activation is critical to the induction of GVHD, and data from our group and others have shown that regulatory T cells (Tregs) prevent GVHD when given at the time of HSCT. Using multiphoton laser scanning microscopy, we examined the single cell dynamics of donor T cells and dendritic cells (DCs) with or without Tregs postallogeneic transplantation. We found that donor conventional T cells (Tcons) spent very little time screening host DCs. Tcons formed stable contacts with DCs very early after transplantation and only increased velocity in the lymph node at 20 hours after transplant. We also observed that Tregs reduced the interaction time between Tcons and DCs, which was dependent on the generation of interleukin 10 by Tregs. Imaging using inducible Tregs showed similar disruption of Tcon-DC contact. Additionally, we found that donor Tregs induce host DC death and down-regulate surface proteins required for donor T-cell activation. These data indicate that Tregs use multiple mechanisms that affect host DC numbers and function to mitigate acute GVHD.

  11. Resonance Enhanced Multi-photon Spectroscopy of DNA

    Science.gov (United States)

    Ligare, Marshall Robert

    For over 50 years DNA has been studied to better understand its connection to life and evolution. These past experiments have led to our understanding of its structure and function in the biological environment but the interaction of DNA with UV radiation at the molecular level is still not very well understood. Unique mechanisms in nucleobase chromaphores protect us from adverse chemical reactions after UV absorption. Studying these processes can help develop theories for prebiotic chemistry and the possibility of alternative forms of DNA. Using resonance enhanced multi-photon spectroscopic techniques in the gas phase allow for the structure and dynamics of individual nucleobases to be studied in detail. Experiments studying different levels of structure/complexity with relation to their biological function are presented. Resonant IR multiphoton dissociation spectroscopy in conjunction with molecular mechanics and DFT calculations are used to determine gas phase structures of anionic nucleotide clusters. A comparison of the identified structures with known biological function shows how the hydrogen bonding of the nucleotides and their clusters free of solvent create favorable structures for quick incorporation into enzymes such as DNA polymerase. Resonance enhanced multi-photon ionization (REMPI) spectroscopy techniques such as resonant two photon ionization (R2PI) and IR-UV double resonance are used to further elucidate the structure and excited state dynamics of the bare nucleobases thymine and uracil. Both exhibit long lived excited electronic states that have been implicated in DNA photolesions which can ultimately lead to melanoma and carcinoma. Our experimental data in comparison with many quantum chemical calculations suggest a new picture for the dynamics of thymine and uracil in the gas phase. A high probability of UV absorption from a vibrationally hot ground state to the excited electronic state shows that the stability of thymine and uracil comes from

  12. Modelling of infrared multiphoton absorption and dissociation for design of reactors for isotope separation by lasers

    International Nuclear Information System (INIS)

    Takeuchi, Kazuo; Nakane, Ryohei; Inoue, Cihiro

    1981-01-01

    A series of experiments were performed on infrared laser beam absorption (multiphoton absorption) and subsequent dissociation (multiphoton dissociation) of CF 3 Cl to propose models for the design of reactors for isotope separation by lasers. A parallel beam geometry was utilized in batch irradiation experiments to make direct compilation of lumped-parameter data possible. Multiphoton absorption is found to be expressed by a power-law extension of the law of Lambert and by an addition of a new term for buffer gas effect to the law of Beer. For reaction analysis, a method to evaluate the effect of incomplete mixing on apparent reaction rates is first presented. Secondly, multiphoton dissociation of Cf 3 Cl is found to occur in pseudo-first order fashion and the specific reaction rates for different beam fluence are shown to be correlated to the absorbed energy. (author)

  13. Multi-photon absorption limits to heralded single photon sources

    Science.gov (United States)

    Husko, Chad A.; Clark, Alex S.; Collins, Matthew J.; De Rossi, Alfredo; Combrié, Sylvain; Lehoucq, Gaëlle; Rey, Isabella H.; Krauss, Thomas F.; Xiong, Chunle; Eggleton, Benjamin J.

    2013-01-01

    Single photons are of paramount importance to future quantum technologies, including quantum communication and computation. Nonlinear photonic devices using parametric processes offer a straightforward route to generating photons, however additional nonlinear processes may come into play and interfere with these sources. Here we analyse spontaneous four-wave mixing (SFWM) sources in the presence of multi-photon processes. We conduct experiments in silicon and gallium indium phosphide photonic crystal waveguides which display inherently different nonlinear absorption processes, namely two-photon (TPA) and three-photon absorption (ThPA), respectively. We develop a novel model capturing these diverse effects which is in excellent quantitative agreement with measurements of brightness, coincidence-to-accidental ratio (CAR) and second-order correlation function g(2)(0), showing that TPA imposes an intrinsic limit on heralded single photon sources. We build on these observations to devise a new metric, the quantum utility (QMU), enabling further optimisation of single photon sources. PMID:24186400

  14. Multiphoton autofluorescence lifetime imaging of induced pluripotent stem cells

    Science.gov (United States)

    Uchugonova, Aisada

    2017-06-01

    The multiphoton fluorescence lifetime imaging tomograph MPTflex with its flexible 360-deg scan head, articulated arm, and tunable femtosecond laser source was employed to study induced pluripotent stem cell (iPS) cultures. Autofluorescence (AF) lifetime imaging was performed with 250-ps temporal resolution and submicron spatial resolution using time-correlated single-photon counting. The two-photon excited AF was based on the metabolic coenzymes NAD(P)H and flavin adenine dinucleotide/flavoproteins. iPS cells generated from mouse embryonic fibroblasts (MEFs) and cocultured with growth-arrested MEFs as feeder cells have been studied. Significant differences on AF lifetime signatures were identified between iPS and feeder cells as well as between their differentiating counterparts.

  15. Observation of multiphoton detachment of the H/sup -/ ion

    International Nuclear Information System (INIS)

    Tang, C.Y.; Harris, P.G.; Mohagheghi, A.H.; Los Alamos National Laboratory, Los Alamos, New Mexico 87545; Cohen Mechanical Design, Broomall, Pennsylvania 19008; The University of Connecticut, Storrs, Connecticut 06268; Western Washington University, Bellingham, Washington 98225)

    1989-01-01

    We have observed nonresonant multiphoton electron detachment of H/sup -/ ions in moderately intense (a few tens of GW/cm 2 ) laser fields. A well-collimated beam of H/sup -/ ions with an energy of 581 MeV was intersected by focused 10.6-μm radiation from a pulsed CO 2 laser. The center-of-mass photon energy was tuned using the relativistic Doppler shift so that the minimum number of simultaneous photons required for electron detachment ranged from three to sixteen. Definite signals were observed for the minimum photon number ranging from three to eight. Our preliminary results show evidence for structure in the relative total cross section

  16. Multiphoton transitions in semiconductors in the non-perturbative approach

    International Nuclear Information System (INIS)

    Iqbal, M.Z.; Hassan, A.R.

    1987-09-01

    Transition rates for multiphoton absorption via direct band-to-band excitation have been calculated using a non-perturbative approach due to Jones and Reiss, based on the Volkov type final state wave functions. Both cases of parabolic and non-parabolic energy bands have been included in our calculations. Absorption coefficients have been obtained for the cases of plane polarized and circularly polarized light. In particular, two-photon absorption coefficients are derived for the two cases of polarization for the parabolic band approximation as well as for non-parabolic bands and compared with the results based on perturbation theory. Numerical estimates of the two photon absorption coefficients resulting from our calculations are also provided. (author). 10 refs, 1 tab

  17. Multiphoton production at high energies in the standard model. II

    International Nuclear Information System (INIS)

    Mahlon, G.

    1993-01-01

    We examine multiphoton production in the electroweak sector of the standard model in the high-energy limit using the equivalence theorem in combination with spinor helicity techniques. We utilize currents consisting of a charged scalar, spinor, or vector line that radiates n photons. Only one end of the charged line is off shell in these currents, which are known for the cases of like-helicity and one unlike-helicity photons. We obtain a wide variety of helicity amplitudes for processes involving two pairs of charged particles by considering combinations of four currents. We examine the situation with respect to currents which have both ends of the charged line off shell, and present solutions for the case of like-helicity photons. These new currents may be combined with two of the original currents to produce additional amplitudes involving Higgs bosons, longitudinal Z, or neutrino pairs

  18. Picosecond multiphoton ionization of atomic and molecular clusters

    International Nuclear Information System (INIS)

    Miller, J.C.; Smith, D.B.

    1990-01-01

    High peak-power picosecond laser pulses have been used for the first time to effect nonresonant or resonant multiphoton ionization (MPI) of clusters generated in a supersonic nozzle expansion. The resulting ions are subsequently detected and characterized by time-of-flight mass spectroscopy. Specifically, we present results involving MPI of clusters of xenon and nitric oxide. Previous MPI studies of many molecular clusters using nanosecond lasers have not been successful in observing the parent ion, presumably due to fast dissociation channels. It is proposed that the present technique is a new and rather general ionization source for cluster studies which is complementary to electron impact but may, in addition, provide unique spectroscopic or dynamical information. 23 refs., 5 figs

  19. Permutational symmetries for coincidence rates in multimode multiphotonic interferometry

    Science.gov (United States)

    Khalid, Abdullah; Spivak, Dylan; Sanders, Barry C.; de Guise, Hubert

    2018-06-01

    We obtain coincidence rates for passive optical interferometry by exploiting the permutational symmetries of partially distinguishable input photons, and our approach elucidates qualitative features of multiphoton coincidence landscapes. We treat the interferometer input as a product state of any number of photons in each input mode with photons distinguished by their arrival time. Detectors at the output of the interferometer count photons from each output mode over a long integration time. We generalize and prove the claim of Tillmann et al. [Phys. Rev. X 5, 041015 (2015), 10.1103/PhysRevX.5.041015] that coincidence rates can be elegantly expressed in terms of immanants. Immanants are functions of matrices that exhibit permutational symmetries and the immanants appearing in our coincidence-rate expressions share permutational symmetries with the input state. Our results are obtained by employing representation theory of the symmetric group to analyze systems of an arbitrary number of photons in arbitrarily sized interferometers.

  20. Monitoring wound healing by multiphoton tomography/endoscopy

    Science.gov (United States)

    König, Karsten; Weinigel, Martin; Bückle, Rainer; Kaatz, Martin; Hipler, Christina; Zens, Katharina; Schneider, Stefan W.; Huck, Volker

    2015-02-01

    Certified clinical multiphoton tomographs are employed to perform rapid label-free high-resolution in vivo histology. Novel tomographs include a flexible 360° scan head attached to a mechano-optical arm for autofluorescence and SHG imaging as well as rigid two-photon GRIN microendoscope. Mitochondrial fluorescent NAD(P)H, fluorescent elastin, keratin, and melanin as well as SHG-active collagen can be imaged with submicron resolution in human skin. The system was employed to study the healing of chronic wounds (venous leg ulcer) and acute wounds (curettage of actinic or seborrheic keratosis) on a subcellular level. Furthermore, a flexible sterile foil as interface between wound and focusing optic was tested.

  1. Multiphoton Rabi oscillations between highly excited Stark states of potassium

    International Nuclear Information System (INIS)

    He Yonglin

    2011-01-01

    We have applied a nonperturbative resonant theory to study the Rabi frequency of microwave multiphoton transitions between two Rydberg states of potassium in a static electric field. The Stark electric dipole moments used to calculate the Rabi frequency are determined by the Stark states' wave functions, which are obtained by the diagonalization method. The frequencies of the Rabi oscillations are in good agreement with either experimental ones or ones calculated by the time-dependent close-coupling method and the Floquet theory. Furthermore, we are able to show that the size of avoided crossings between the (n+2)s and (n,3) states can be predicted from the Stark electric dipole moment and the difference of the two Stark states' energy at a given resonance.

  2. Search for Multiphoton Signatures of a Higgs Boson

    Energy Technology Data Exchange (ETDEWEB)

    Atramentov, Oleksiy Vladimirovich [Iowa State Univ., Ames, IA (United States)

    2006-07-01

    In this thesis we describe a search for a fermiophobic Higgs boson in 3γ+X events. The study has been performed on 0.83 fb-1 of data collected with the D0 detector that resides at one of the interaction regions of the Tevatron collider, the world's highest energy accelerator. This study was motivated by a fairly recent phenomenological paper [33] where it was noticed that in certain class of models (2HDM Type I and THM) the multi-photon final states like this one become detectable at the luminosity that has been collected by the D0 experiment by 2006. The mechanism that permits such final state becomes available when the conventional higgs production mechanism (higgs strahlung) are suppressed. This leads to the fact that Higgs boson with mass (mhf < 90 GeV/c2) lower than the current limit has not been excluded.

  3. Variational methods for high-order multiphoton processes

    International Nuclear Information System (INIS)

    Gao, B.; Pan, C.; Liu, C.; Starace, A.F.

    1990-01-01

    Methods for applying the variationally stable procedure for Nth-order perturbative transition matrix elements of Gao and Starace [Phys. Rev. Lett. 61, 404 (1988); Phys. Rev. A 39, 4550 (1989)] to multiphoton processes involving systems other than atomic H are presented. Three specific cases are discussed: one-electron ions or atoms in which the electron--ion interaction is described by a central potential; two-electron ions or atoms in which the electronic states are described by the adiabatic hyperspherical representation; and closed-shell ions or atoms in which the electronic states are described by the multiconfiguration Hartree--Fock representation. Applications are made to the dynamic polarizability of He and the two-photon ionization cross section of Ar

  4. Clinical multiphoton tomography and clinical two-photon microendoscopy

    Science.gov (United States)

    König, Karsten; Bückle, Rainer; Weinigel, Martin; Elsner, Peter; Kaatz, Martin

    2009-02-01

    We report on applications of high-resolution clinical multiphoton tomography based on the femtosecond laser system DermaInspectTM with its flexible mirror arm in Australia, Asia, and Europe. Applications include early detection of melanoma, in situ tracing of pharmacological and cosmetical compounds including ZnO nanoparticles in the epidermis and upper dermis, the determination of the skin aging index SAAID as well as the study of the effects of anti-aging products. In addition, first clinical studies with novel rigid high-NA two-photon 1.6 mm GRIN microendoscopes have been conducted to study the effect of wound healing in chronic wounds (ulcus ulcera) as well as to perform intrabody imaging with subcellular resolution in small animals.

  5. One color multi-photon ionization of the Gadolinium atom in near UV region

    International Nuclear Information System (INIS)

    Kim, Jin Tae; Yi, Jong Hoon; Lhee, Yong Joo; Lee, Jong Min

    1999-01-01

    We have investigated the states of the gadolinium atom in near ultra-violet (UV) region (∼410 nm) using single photon excitation using resonance ionization mass spectrometry (RIMS). Around 70 transitions among observed 180 single color multi-photon ionization signals have been assigned. Most of the multi-photon processes of the assigned ion signals are through single photon resonant three photon ionization and through two photon resonant three photon ionization. (author)

  6. Recoil effects in multiphoton electron-positron pair creation

    International Nuclear Information System (INIS)

    Krajewska, K.; Kaminski, J. Z.

    2010-01-01

    Triply differential probability rates for electron-positron pair creation in laser-nucleus collisions, calculated within the S-matrix approach, are investigated as functions of the nuclear recoil. Pronounced enhancements of differential probability rates of multiphoton pair production are found for a nonzero momentum transfer from the colliding nucleus. The corresponding rates show a very dramatic dependence on the polarization of the laser field impinging on the nucleus; only for a linearly polarized light are the multiphoton rates for electron-positron pair production considerably large. We focus therefore on this case. Our numerical results for different geometries of the reaction particles demonstrate that, for the linearly polarized laser field of an infinite extent (which is a good approximation for femtosecond laser pulses), the pair creation is far more efficient if the nucleus is detected in the direction of the laser-field propagation. The corresponding angular distributions of the created particles show that the high-energy pairs are predominantly produced in the plane spanned by the polarization vector and the laser-field propagation direction, while the low-energy pairs are rather spread around the latter of the two directions. The enhancement of differential probability rates at each energy sector, defined by the four-momentum conservation relation, is observed with varying the energy of the produced particles. The total probability rates of pair production are also evaluated and compared with the corresponding results for the case when one disregards the recoil effect. A tremendous enhancement of the total probability rates of the electron-positron pair creation is observed if one takes into account the nuclear recoil.

  7. An enhancement of spin polarization by multiphoton pumping in semiconductors

    Energy Technology Data Exchange (ETDEWEB)

    Miah, M. Idrish, E-mail: m.miah@griffith.edu.au [Queensland Micro- and Nanotechnology Centre, Griffith University, Nathan, Brisbane, QLD 4111 (Australia); Department of Physics, University of Chittagong, Chittagong 4331 (Bangladesh)

    2011-08-15

    Highlights: {yields} Multiphoton pumping and spin generation in semiconductors. {yields} Optical selection rules for inter-band transitions. {yields} Calculations of spin polarization using band-energy model and the second order perturbation theory. {yields} Enhancement of the electronic spin polarization. - Abstract: A pump-probe spectroscopic study has been carried out in zinc-blende bulk semiconductors. In the semiconductor samples, a spin-polarized carrier population is produced by the absorption of a monochromatic circularly polarized light beam with two-photon energy above the direct band gap in bulk semiconductors. The production of a carrier population with a net spin is a consequence of the optical selection rules for the heavy-hole and light-hole valence-to-conduction band transitions. This production is probed by the spin-dependent transmission of the samples in the time domain. The spin polarization of the conduction-band-electrons in dependences of delay of the probe beam as well as of pumping photon energy is estimated. The spin polarization is found to depolarize rapidly for pumping energy larger than the energy gap of the split-off band to the conduction band. From the polarization decays, the spin relaxation times are also estimated. Compared to one-photon pumping, the results, however, show that an enhancement of the spin-polarization is achieved by multiphoton excitation of the samples. The experimental results are compared with those obtained in calculations using second order perturbation theory of the spin transport model. A good agreement between experiment and theory is obtained. The observed results are discussed in details.

  8. An enhancement of spin polarization by multiphoton pumping in semiconductors

    International Nuclear Information System (INIS)

    Miah, M. Idrish

    2011-01-01

    Highlights: → Multiphoton pumping and spin generation in semiconductors. → Optical selection rules for inter-band transitions. → Calculations of spin polarization using band-energy model and the second order perturbation theory. → Enhancement of the electronic spin polarization. - Abstract: A pump-probe spectroscopic study has been carried out in zinc-blende bulk semiconductors. In the semiconductor samples, a spin-polarized carrier population is produced by the absorption of a monochromatic circularly polarized light beam with two-photon energy above the direct band gap in bulk semiconductors. The production of a carrier population with a net spin is a consequence of the optical selection rules for the heavy-hole and light-hole valence-to-conduction band transitions. This production is probed by the spin-dependent transmission of the samples in the time domain. The spin polarization of the conduction-band-electrons in dependences of delay of the probe beam as well as of pumping photon energy is estimated. The spin polarization is found to depolarize rapidly for pumping energy larger than the energy gap of the split-off band to the conduction band. From the polarization decays, the spin relaxation times are also estimated. Compared to one-photon pumping, the results, however, show that an enhancement of the spin-polarization is achieved by multiphoton excitation of the samples. The experimental results are compared with those obtained in calculations using second order perturbation theory of the spin transport model. A good agreement between experiment and theory is obtained. The observed results are discussed in details.

  9. Multiphoton dissociation and thermal unimolecular reactions induced by infrared lasers

    International Nuclear Information System (INIS)

    Dai, H.L.

    1981-04-01

    Multiphoton dissociation (MPD) of ethyl chloride was studied using a tunable 3.3 μm laser to excite CH stretches. The absorbed energy increases almost linearly with fluence, while for 10 μm excitation there is substantial saturation. Much higher dissociation yields were observed for 3.3 μm excitation than for 10 μm excitation, reflecting bottlenecking in the discrete region of 10 μm excitation. The resonant nature of the excitation allows the rate equations description for transitions in the quasicontinuum and continuum to be extended to the discrete levels. Absorption cross sections are estimated from ordinary ir spectra. A set of cross sections which is constant or slowly decreasing with increasing vibrational excitation gives good fits to both absorption and dissociation yield data. The rate equations model was also used to quantitatively calculate the pressure dependence of the MPD yield of SF 6 caused by vibrational self-quenching. Between 1000-3000 cm -1 of energy is removed from SF 6 excited to approx. > 60 kcal/mole by collision with a cold SF 6 molecule at gas kinetic rate. Calculation showed the fluence dependence of dissociation varies strongly with the gas pressure. Infrared multiphoton excitation was applied to study thermal unimolecular reactions. With SiF 4 as absorbing gas for the CO 2 laser pulse, transient high temperature pulses were generated in a gas mixture. IR fluorescence from the medium reflected the decay of the temperature. The activation energy and the preexponential factor of the reactant dissociation were obtained from a phenomenological model calculation. Results are presented in detail

  10. Advances in polarization sensitive multiphoton nano-bio-imaging

    Directory of Open Access Journals (Sweden)

    Zyss J.

    2010-06-01

    Full Text Available In this talk, we shall shortly review four main directions of ongoing research in our laboratories, directed at the conception and demonstration of a variety of innovative configurations in nanoscale multiphoton imaging. A common feature to all of these directions appears to be the central role played by the involvement of polarization features, both in- and outgoing, moreover so in view of the tensorial aspects inherent to nonlinear schemes such second-harmonic generation, electro-optic modulation or two-photon fluorescence which will ne emphasized. These advances relate to the new domain of nonlinear ellipsometry in multiphoton imaging [1], of high relevance to fundamental aspects of nanophotonics and nanomaterial engineering as well as towards basic life science issues. The four domains to be shortly reported are: a polarization resolved second-harmonic generation in semiconductor QD’s with record small sizes in the 10-12 nm range [2] b original use of two-photon confocal polarization resolved microscopy in DNA stained by two photon fluorescent dyes in different LC phases arrangements so as to characterize these as well as ascertain the respective DNA-dye orientation (intercalant or groves [3] c elaboration and demonstration of an electrooptic confocal microscope in a highly sensitive interferometric and homodyne detection configuration allowing to map weak electric potentials such as in artificial functionalized membranes, the dynamical investigation of firing and propagation aspects of action potentials in neurones being currently the next step [4] d original plasmon based enhanced nanoscale confocal imaging involving a dual detection scheme (fluorescence imaging and ATR plasmon coupling in reflection whereby adequate preparation and switching of the incoming polarization state between radial, linear and azimuthal configurations, entail different images and plasmon enhancement levels [5].

  11. Complementary roles of platelets and coagulation in thrombus formation on plaques acutely ruptured by targeted ultrasound treatment: a novel intravital model

    NARCIS (Netherlands)

    Kuijpers, M. J. E.; Gilio, K.; Reitsma, S.; Nergiz-Unal, R.; Prinzen, L.; Heeneman, S.; Lutgens, E.; van Zandvoort, M. A. M. J.; Nieswandt, B.; Egbrink, M. G. A. Oude; Heemskerk, J. W. M.

    2009-01-01

    Atherothrombosis is a major cause of cardiovascular events. However, animal models to study this process are scarce. We describe the first murine model of acute thrombus formation upon plaque rupture to study atherothrombosis by intravital fluorescence microscopy. Localized rupture of an

  12. Contribution of laser Doppler flowmetry with venoarteriolar reflex, cold, and rewarming testing, and intravital capillaroscopy to diagnose Raynaud's phenomenon

    Directory of Open Access Journals (Sweden)

    Zeman J

    2014-05-01

    Full Text Available Jan Zeman,1 Oksana Turyanytsya,1 Vojtĕch Kapsa,2 Mojmír Eliáš3 1Department of Clinical Cardiology and Angiology, Hospital Bulovka, 2Charles University in Prague, Faculty of Mathematics and Physics, 3Kooperativa a.s., Pobrezni, Prague, Czech Republic Background: The early differential diagnosis of Raynaud’s phenomenon (RP is crucial for the prognosis and therapy of these patients. In our microcirculatory laboratory, we use intravital capillaroscopy (IC, plethysmography (P, and laser Doppler flowmetry (LDF for examining acrosyndromes. We combine LDF with venoarteriolar reflex test, cold test, and rewarming test to achieve more reliable diagnoses of acrosyndromes. Patients and methods: We examined LDF and IC according to a strict protocol using a battery of tests (venoarteriolar reflex test, cold test, rewarming test applied to five different groups of people and compared their results: healthy controls, primary Raynaud’s phenomenon (PRP, systemic scleroderma, vibration white finger, and peripheral artery occlusive disease. Our tests included 340 individuals (72 patients plus 268 controls. Results: Although all tests provided some differences between controls and patients, only the rewarming test offered significant results for differential diagnoses. Conclusion: IC and LDF combined with the battery of tests (venoarteriolar reflex test, cold test, rewarming test under standard conditions can be used as reliable tools to distinguish between PRP and some types of secondary RP (especially in the case of systemic scleroderma, vibration white fingers, or peripheral artery occlusive disease; RPs with organic occlusions of the small arteries causing the diseases. Our methodology can help to distinguish between other types of RP, as well. Keywords: Raynaud’s phenomenon, acrosyndrome, laser Doppler flowmetry, intravital capillaroscopy, scleroderma, vibration white finger, peripheral artery occlusive disease

  13. An orthotopic glioblastoma mouse model maintaining brain parenchymal physical constraints and suitable for intravital two-photon microscopy.

    Science.gov (United States)

    Ricard, Clément; Stanchi, Fabio; Rougon, Geneviève; Debarbieux, Franck

    2014-04-21

    Glioblastoma multiforme (GBM) is the most aggressive form of brain tumors with no curative treatments available to date. Murine models of this pathology rely on the injection of a suspension of glioma cells into the brain parenchyma following incision of the dura-mater. Whereas the cells have to be injected superficially to be accessible to intravital two-photon microscopy, superficial injections fail to recapitulate the physiopathological conditions. Indeed, escaping through the injection tract most tumor cells reach the extra-dural space where they expand abnormally fast in absence of mechanical constraints from the parenchyma. Our improvements consist not only in focally implanting a glioma spheroid rather than injecting a suspension of glioma cells in the superficial layers of the cerebral cortex but also in clogging the injection site by a cross-linked dextran gel hemi-bead that is glued to the surrounding parenchyma and sealed to dura-mater with cyanoacrylate. Altogether these measures enforce the physiological expansion and infiltration of the tumor cells inside the brain parenchyma. Craniotomy was finally closed with a glass window cemented to the skull to allow chronic imaging over weeks in absence of scar tissue development. Taking advantage of fluorescent transgenic animals grafted with fluorescent tumor cells we have shown that the dynamics of interactions occurring between glioma cells, neurons (e.g. Thy1-CFP mice) and vasculature (highlighted by an intravenous injection of a fluorescent dye) can be visualized by intravital two-photon microscopy during the progression of the disease. The possibility to image a tumor at microscopic resolution in a minimally compromised cerebral environment represents an improvement of current GBM animal models which should benefit the field of neuro-oncology and drug testing.

  14. The mouse cortical meninges are the site of immune responses to many different pathogens, and are accessible to intravital imaging.

    Science.gov (United States)

    Coles, Jonathan A; Stewart-Hutchinson, Phillip J; Myburgh, Elmarie; Brewer, James M

    2017-08-15

    A wide range of viral and microbial infections are known to cause meningitis, and there is evidence that the meninges are the gateway to pathogenic invasion of the brain parenchyma. Hence observation of these regions has wide application to understanding host-pathogen interactions. Interactions between pathogens and cells of the immune response can be modified by changes in their environment, such as suppression of the flow of blood and lymph, and, particularly in the case of the meninges, with their unsupported membranes, invasive dissection can alter the tissue architecture. For these reasons, intravital imaging through the unperforated skull is the method of choice. We give a protocol for a simple method of two-photon microscopy through the thinned cortical skull of the anesthetized mouse to enable real-time imaging with sub-micron resolution through the meninges and into the superficial brain parenchyma. In reporter mice in which selected cell types express fluorescent proteins, imaging after infection with fluorescent pathogens (lymphocytic choriomeningitis virus, Trypanosoma brucei or Plasmodium berghei) has shown strong recruitment to the cortical meninges of immune cells, including neutrophils, T cells, and putative dendritic cells and macrophages. Without special labeling, the boundaries between the dura mater, the leptomeninx, and the parenchyma are not directly visualized in intravital two-photon microscopy, but other landmarks and characteristics, which we illustrate, allow the researcher to identify the compartment being imaged. While most infectious meningitides are localized mainly in the dura mater, others involve recruitment of immune cells to the leptomeninx. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  15. Multiphoton (e,2e) process of hydrogen atom in strong laser field

    Energy Technology Data Exchange (ETDEWEB)

    Ghosh Deb, S.; Roy, S.; Sinha, C. [Indian Association for the Cultivation of Science, Dept. of Theoretical Physics, Jadavpur, Kolkata (India)

    2009-12-15

    The dynamics of the electron impact multiphoton ionization of a hydrogen atom in the presence of an intense laser field (e, n gamma e) has been studied theoretically for laser polarization parallel and perpendicular to the incident momentum, with a view to comparing (qualitatively) the results with the recent kinematically complete experiments of Hoehr et al. for the He target. Significant laser modifications are noted in the present doubly (DDCS) and the fully differential multiphoton cross sections (TDCS) for both the geometries (parallel and perpendicular). For most of the explored kinematics (chosen in accordance with the experiment), the present binary peak intensity of the laser-assisted multiphoton TDCS is significantly enhanced with respect to the field free ones, in qualitative agreement with the experiment. Importance of the multiphoton effects is also studied. The multiphoton cross sections in the zeroth order approximation of the ejected electron wavefunction (CV) obeys the Kroll Watson sum rule while it does not hold good in the corresponding first order approximation (MCV). (authors)

  16. Structure of multiphoton quantum optics. I. Canonical formalism and homodyne squeezed states

    Science.gov (United States)

    dell'Anno, Fabio; de Siena, Silvio; Illuminati, Fabrizio

    2004-03-01

    We introduce a formalism of nonlinear canonical transformations for general systems of multiphoton quantum optics. For single-mode systems the transformations depend on a tunable free parameter, the homodyne local-oscillator angle; for n -mode systems they depend on n heterodyne mixing angles. The canonical formalism realizes nontrivial mixing of pairs of conjugate quadratures of the electromagnetic field in terms of homodyne variables for single-mode systems, and in terms of heterodyne variables for multimode systems. In the first instance the transformations yield nonquadratic model Hamiltonians of degenerate multiphoton processes and define a class of non-Gaussian, nonclassical multiphoton states that exhibit properties of coherence and squeezing. We show that such homodyne multiphoton squeezed states are generated by unitary operators with a nonlinear time evolution that realizes the homodyne mixing of a pair of conjugate quadratures. Tuning of the local-oscillator angle allows us to vary at will the statistical properties of such states. We discuss the relevance of the formalism for the study of degenerate (up-)down-conversion processes. In a companion paper [ F. Dell’Anno, S. De Siena, and F. Illuminati, 69, 033813 (2004) ], we provide the extension of the nonlinear canonical formalism to multimode systems, we introduce the associated heterodyne multiphoton squeezed states, and we discuss their possible experimental realization.

  17. Multiphoton electronic-spin generation and transmission spectroscopy in n-type GaAs

    Energy Technology Data Exchange (ETDEWEB)

    Idrish Miah, M., E-mail: m.miah@griffith.edu.a [Department of Physics, University of Chittagong, Chittagong 4331 (Bangladesh)

    2011-01-17

    Multiphoton electronic-spin generation in semiconductors was investigated using differential transmission spectroscopy. The generation of the electronic spins in the semiconductor samples were achieved by multiphoton pumping with circularly polarized light beam and was probed by the spin-resolved transmission of the samples. The electronic spin-polarization of conduction band was estimated and was found to depend on the delay of the probe beam, temperature as well as on the multiphoton pumping energy. The temperature dependence showed a decrease of the spin-polarization with increasing temperature. The electronic spin-polarization was found to depolarize rapidly for multiphoton pumping energy larger than the energy gap of the split-off band to the conduction band. The results were compared with those obtained in one-photon pumping, which shows that an enhancement of the electronic spin-polarization was achieved in multiphoton pumping. The findings resulting from this investigation might have potential applications in opto-spintronics, where the generation of highly polarized electronic spins is required.

  18. Multiphoton electronic-spin generation and transmission spectroscopy in n-type GaAs

    International Nuclear Information System (INIS)

    Idrish Miah, M.

    2011-01-01

    Multiphoton electronic-spin generation in semiconductors was investigated using differential transmission spectroscopy. The generation of the electronic spins in the semiconductor samples were achieved by multiphoton pumping with circularly polarized light beam and was probed by the spin-resolved transmission of the samples. The electronic spin-polarization of conduction band was estimated and was found to depend on the delay of the probe beam, temperature as well as on the multiphoton pumping energy. The temperature dependence showed a decrease of the spin-polarization with increasing temperature. The electronic spin-polarization was found to depolarize rapidly for multiphoton pumping energy larger than the energy gap of the split-off band to the conduction band. The results were compared with those obtained in one-photon pumping, which shows that an enhancement of the electronic spin-polarization was achieved in multiphoton pumping. The findings resulting from this investigation might have potential applications in opto-spintronics, where the generation of highly polarized electronic spins is required.

  19. Application of multiphoton microscopy in dermatological studies: A mini-review

    Directory of Open Access Journals (Sweden)

    Elijah Yew

    2014-09-01

    Full Text Available This review summarizes the historical and more recent developments of multiphoton microscopy, as applied to dermatology. Multiphoton microscopy offers several advantages over competing microscopy techniques: there is an inherent axial sectioning, penetration depths that compete well with confocal microscopy on account of the use of near-infrared light, and many two-photon contrast mechanisms, such as second-harmonic generation, have no analogue in one-photon microscopy. While the penetration depths of photons into tissue are typically limited on the order of hundreds of microns, this is of less concern in dermatology, as the skin is thin and readily accessible. As a result, multiphoton microscopy in dermatology has generated a great deal of interest, much of which is summarized here. The review covers the interaction of light and tissue, as well as the various considerations that must be made when designing an instrument. The state of multiphoton microscopy in imaging skin cancer and various other diseases is also discussed, along with the investigation of aging and regeneration phenomena, and finally, the use of multiphoton microscopy to analyze the transdermal transport of drugs, cosmetics and other agents is summarized. The review concludes with a look at potential future research directions, especially those that are necessary to push these techniques into widespread clinical acceptance.

  20. Structure of multiphoton quantum optics. I. Canonical formalism and homodyne squeezed states

    International Nuclear Information System (INIS)

    Dell'Anno, Fabio; De Siena, Silvio; Illuminati, Fabrizio

    2004-01-01

    We introduce a formalism of nonlinear canonical transformations for general systems of multiphoton quantum optics. For single-mode systems the transformations depend on a tunable free parameter, the homodyne local-oscillator angle; for n-mode systems they depend on n heterodyne mixing angles. The canonical formalism realizes nontrivial mixing of pairs of conjugate quadratures of the electromagnetic field in terms of homodyne variables for single-mode systems, and in terms of heterodyne variables for multimode systems. In the first instance the transformations yield nonquadratic model Hamiltonians of degenerate multiphoton processes and define a class of non-Gaussian, nonclassical multiphoton states that exhibit properties of coherence and squeezing. We show that such homodyne multiphoton squeezed states are generated by unitary operators with a nonlinear time evolution that realizes the homodyne mixing of a pair of conjugate quadratures. Tuning of the local-oscillator angle allows us to vary at will the statistical properties of such states. We discuss the relevance of the formalism for the study of degenerate (up-)down-conversion processes. In a companion paper [F. Dell'Anno, S. De Siena, and F. Illuminati, 69, 033813 (2004)], we provide the extension of the nonlinear canonical formalism to multimode systems, we introduce the associated heterodyne multiphoton squeezed states, and we discuss their possible experimental realization

  1. Two-photon emission and multiphoton absorption by atoms

    International Nuclear Information System (INIS)

    Mu, X.

    1988-01-01

    This thesis consists of investigations of two problems concerning photon-atom interactions. The first topic deals with two-photon transitions in atomic inner shells. An independent-particle model has been used to describe the two-photon transitions between different inner-shell electron states. The first relativistic self-consistent-field calculation of these transition rates in Ag, Mo, and Xe has been carried out. The theoretical results are compared with recent measurements. Good agreement with measured rates is found except in some cases where more reliable experiments still need to be done. The second topic is multiphoton multiionization of atoms. The maximum entropy principle has been employed in this theoretical investigation. A detailed statistical analysis of measured ionic charge distributions produced in strong laser pulses has been carried out. The results of this analysis indicates that the charge-state distribution is a Poissonian, rather than the binomial which prevails under infrared radiation, and hence that ionization occurs stepwise during the pulse. This result is shown to be consistent with experimental data

  2. Multiphoton gonioscopy to image the trabecular meshwork of porcine eyes

    Science.gov (United States)

    Masihzadeh, Omid; Ammar, David A.; Kahook, Malik Y.; Gibson, Emily A.; Lei, Tim C.

    2013-03-01

    The aqueous outflow system (AOS), including the trabecular meshwork (TM), the collector channels (CC) and the Schlemm's canal (SC), regulates intraocular pressure (IOP) through the drainage of the aqueous humor (AH). Abnormal IOP elevation leads to increased pressure stress to retinal ganglion cells, resulting in cell loss that can ultimately lead to complete loss of eyesight. Therefore, development of imaging tools to detect abnormal structural and functional changes of the AOS is important in early diagnosis and prevention of glaucoma. Multiphoton microscopy (MPM), including twophoton autofluorescence (TPAF) and second harmonic generation (SHG), is a label-free microscopic technique that allows molecular specific imaging of biological tissues like the TM. Since the TM and other AOS structures are located behind the highly scattering scleral tissue, transscleral imaging of the TM does not provide enough optical resolution. In this work, a gonioscopic lens is used to allow direct optical access of the TM through the cornea for MPM imaging. Compared to transscleral imaging, the acquired MPM images show improved resolution as individual collagen fiber bundles of the TM can be observed. MPM gonioscopy may have the potential to be developed as a future clinical imaging tool for glaucoma diagnostics.

  3. Direct trabecular meshwork imaging in porcine eyes through multiphoton gonioscopy

    Science.gov (United States)

    Masihzadeh, Omid; Ammar, David A.; Kahook, Malik Y.; Gibson, Emily A.; Lei, Tim C.

    2013-03-01

    The development of technologies to characterize the ocular aqueous outflow system (AOS) is important for the understanding of the pathophysiology of glaucoma. Multiphoton microscopy (MPM) offers the advantage of high-resolution, label-free imaging with intrinsic image contrast because the emitted signals result from the specific biomolecular content of the tissue. Previous attempts to use MPM to image the murine irido-corneal region directly through the sclera have suffered from degradation in image resolution due to scattering of the focused laser light. As a result, transscleral MPM has limited ability to observe fine structures in the AOS. In this work, the porcine irido-corneal angle was successfully imaged through the transparent cornea using a gonioscopic lens to circumvent the highly scattering scleral tissue. The resulting high-resolution images allowed the detailed structures in the trabecular meshwork (TM) to be observed. Multimodal imaging by two-photon autofluorescence and second harmonic generation allowed visualization of different features in the TM without labels and without disruption of the TM or surrounding tissues. MPM gonioscopy is a promising noninvasive imaging tool for high-resolution studies of the AOS, and research continues to explore the potential for future clinical applications in humans.

  4. Development of resonance-enhanced multiphoton ionization system

    International Nuclear Information System (INIS)

    Naik, P.D.; Upadhyaya, Hari P.; Kumar, Awadhesh; Bajaj, P.N.; Sinha, A.K.; Bhatt, S.; Gupta, M.D.P.

    2009-05-01

    Radiation and Photochemistry Division has developed a Molecular Beam-Resonance Enhanced Multiphoton Ionization-Time-of-Flight spectrometer, a highly sensitive and selective analytical detection system, for investigation of photodissociation dynamics of isolated molecules. In this system, the molecular beam is intersected in the extraction region of a Wiley-McLaren type Time-of-Flight mass spectrometer by the photolysis laser beam, propagating perpendicular to both the molecular beams and the Time-of-Flight tube. The probe (ionization) laser beam counter propagating to the photolysis beam, ionizes the stable products and the radicals produced on photodissociation. The important features of the system, namely, the resolution and the detection limit, have been determined from the studies of aniline molecular beam, generated by seeding 1% aniline in helium. For the present configuration, using one metre long flight tube, the resolution has been found to be about 400, and detection limit is better than 106 species per cm 3 . The integrity of the set-up is obtained from the photodissociation dynamics studies of bromoform. (author)

  5. Multiphoton ionization photoelectron spectroscopy of xenon: Experiment and theory

    International Nuclear Information System (INIS)

    Bajic, S.J.; Compton, R.N.; Tang, X.; L'Huiller, A.; Lambropoulos, P.

    1988-11-01

    Photoelectron energy and angular distributions for resonantly enhanced multiphoton ionization (REMPI) of xenon via the three-photon-allowed 7s[3/2] 1 0 and 5d[3/2] 1 0 states have been studied both experimentally and theoretically. The electron kinetic energy spectra give the probability of leaving Xe + in either the 2 P/sub 1/2/ or 2 P/sub 3/2/ core. The measured branching ratio for leaving each ionic core is used to test the theoretical description of the REMPI process. Measurements of both the angular distributions and the [3+1] REMPI via the 5d state are adequately reproduced by multichannel quantum defect theory. However, measurements of angular distributions for the electrons resulting from [3+1] via the 7s[3/2] 1 0 state into Xe + 2 P/sub 3/2/ (core preserving) or Xe + 2 P/sub 1/2/ (core changing) are in striking disagreement with theory. 1 ref., 2 figs

  6. Photoionization of excited molecular states using multiphoton excitation techniques

    International Nuclear Information System (INIS)

    Dehmer, P.M.; Pratt, S.T.; Dehmer, J.L.

    1984-01-01

    Photoelectron spectra are reported for three photon resonant, four photon ionization of H 2 via the B 1 Σ/sub u/ + , v = 7 (J = 2,4) and C 1 π/sub u'/, v = 0-4 (J = 1) levels and of N 2 via the o 3 1 π/sub u'/, v = 1,2, b 1 π/sub u'/, v = 3-5, and c 1 π/sub u'/, v = 0 levels. The results reflect both the spectroscopy and the dynamics of photoionization of excited molecular states and are discussed in terms of the selection rules for photoionization and the relative probabilities of photoionization from Rydberg and valence states. In some cases, in accordance with the Franck-Condon principle, the results demonstrate that resonant multiphoton ionization through Rydberg states may be a powerful technique for the production of electronic, vibrational, and rotational state selected ions. However, in other cases, systematic departures from Franck-Condon factors are observed, which reflect the more subtle dynamics of excited state photoionization. 23 references, 6 figures, 2 tables

  7. Photoionization of excited molecular states using multiphoton excitation techniques

    International Nuclear Information System (INIS)

    Dehmer, P.M.; Pratt, S.T.; Dehmer, J.L.

    1984-01-01

    Photoelectron spectra are reported for three photon resonant, four photon ionization of H 2 via the B 1 Σ + /sub u/, v = 7 (J = 2,4) and C 1 Pi/sub u/, v = 0-4 (J = 1) levels and of N 2 via the o 3 1 Pi/sub u/, v = 1,2, b 1 Pi/sub u/, v = 3-5, and c 1 Pi/sub u/, v = 0 levels. The results reflect both the spectroscopy and the dynamics of photoionization of excited molecular states and are discussed in terms of the selection rules for photoionization and the relative probabilities of photoionization from Rydberg and valence states. In some cases, in accordance with the Franck-Condon principle, the results demonstrate that resonant multiphoton ionization through Rydberg states may be a powerful technique for the production of electronic, vibrational, and rotational state selected ions. However, in other cases, systematic departures from Franck-Condon factors are observed, which reflect the more subtle dynamics of excited state photoionization

  8. Identification of intramural metastasis in esophageal cancer using multiphoton microscopy

    Science.gov (United States)

    Xu, Jian; Kang, Deyong; Zhuo, Shuangmu; Zhu, Xiaoqin; Lin, jiangbo; Chen, Jianxin

    2017-02-01

    Intramural metastasis (IM) of esophageal cancer is defined as metastasis from a primary lesion to the esophageal wall without intraepithelial cancer extension. Esophageal cancer with IM is more common and such cases indicate a poor prognosis. In esophageal surgery, if curative resection is possible, the complete removal of both primary tumor and associated IMs is required. Therefore, accurate diagnosis of IMs in esophageal cancer prior to surgery is of particular importance. Multiphoton microscopy (MPM) with subcellular resolution is well-suited for deep tissue imaging since many endogenous fluorophores of fresh biological tissues are excited through two-photon excited fluorescence (TPEF) and second harmonic generation (SHG). Here, a study to identify IM in fresh tissue section using MPM is reported. In this study, the morphological and spectral differences between IM and surrounding tissue are described. These results show that MPM has the ability to accurately identify IM in esophageal tissues. With improvement of the penetration depth of MPM and the development of multiphton microendoscope, MPM may be a promising imaging technique for preoperative diagnosis of IMs in esophageal cancer in the future.

  9. Multiphoton Ionization Detection in Collinear Laser Spectroscopy of Isolde Beams

    CERN Multimedia

    2002-01-01

    The experiments using the multiphoton ionization technique have been continued in the beginning of 1990 with stable beam tests on the modified apparatus and with another radioactive beam time on Yb. Higher laser power and an increased vacuum in the ionization region (see figure) yielded a further gain in sensitivity, mainly due to the better suppression of the background ions produced in rest gas collisions. For even Yb isotopes we have now reached a detection efficiency of $\\epsilon$~=~1~x~10$^{-5}$ ions per incoming atom at a background count rate of 30~ions from a beam of 5~x~10$^9$. This sensitivity was high enough for spectroscopy on $^{157}$Yb, where the typical ISOLDE yield of 5~x~10$^7$Yb ions is covered by an isobaric contamination of more than 10$^{10}$ ions. Measurements have also been performed on $^{175}$Yb. These give the first precise value for the magnetic moment of this isotope, $\\mu$~=~0.766(8)$ mu _{N} $, which agrees rather well with the magnetic moment of the isotone $^{177}$Hf. The isoto...

  10. Dissecting multi-photon resonances at the large hadron collider

    Energy Technology Data Exchange (ETDEWEB)

    Allanach, B.C. [University of Cambridge, Department of Applied Mathematics and Theoretical Physics, Centre for Mathematical Sciences, Cambridge (United Kingdom); Bhatia, D.; Iyer, Abhishek M. [Tata Institute of Fundamental Research, Department of Theoretical Physics, Mumbai (India)

    2017-09-15

    We examine the phenomenology of the production, at the 13 TeV Large Hadron Collider (LHC), of a heavy resonance X, which decays via other new on-shell particles n into multi-(i.e. three or more) photon final states. In the limit that n has a much smaller mass than X, the multi-photon final state may dominantly appear as a two-photon final state because the γs from the n decay are highly collinear and remain unresolved. We discuss how to discriminate this scenario from X → γγ: rather than discarding non-isolated photons, it is better to relax the isolation criteria and instead form photon jets substructure variables. The spins of X and n leave their imprint upon the distribution of pseudo-rapidity gap Δη between the apparent two-photon states. Depending on the total integrated luminosity, this can be used in many cases to claim discrimination between the possible spin choices of X and n, although the case where X and n are both scalar particles cannot be discriminated from the direct X → γγ decay in this manner. Information on the mass of n can be gained by considering the mass of each photon jet. (orig.)

  11. Adaptive optics improves multiphoton super-resolution imaging

    Science.gov (United States)

    Zheng, Wei; Wu, Yicong; Winter, Peter; Shroff, Hari

    2018-02-01

    Three dimensional (3D) fluorescence microscopy has been essential for biological studies. It allows interrogation of structure and function at spatial scales spanning the macromolecular, cellular, and tissue levels. Critical factors to consider in 3D microscopy include spatial resolution, signal-to-noise (SNR), signal-to-background (SBR), and temporal resolution. Maintaining high quality imaging becomes progressively more difficult at increasing depth (where optical aberrations, induced by inhomogeneities of refractive index in the sample, degrade resolution and SNR), and in thick or densely labeled samples (where out-of-focus background can swamp the valuable, in-focus-signal from each plane). In this report, we introduce our new instrumentation to address these problems. A multiphoton structured illumination microscope was simply modified to integrate an adpative optics system for optical aberrations correction. Firstly, the optical aberrations are determined using direct wavefront sensing with a nonlinear guide star and subsequently corrected using a deformable mirror, restoring super-resolution information. We demonstrate the flexibility of our adaptive optics approach on a variety of semi-transparent samples, including bead phantoms, cultured cells in collagen gels and biological tissues. The performance of our super-resolution microscope is improved in all of these samples, as peak intensity is increased (up to 40-fold) and resolution recovered (up to 176+/-10 nm laterally and 729+/-39 nm axially) at depths up to 250 μm from the coverslip surface.

  12. Visualizing liver anatomy, physiology and pharmacology using multiphoton microscopy.

    Science.gov (United States)

    Wang, Haolu; Liang, Xiaowen; Gravot, Germain; Thorling, Camilla A; Crawford, Darrell H G; Xu, Zhi Ping; Liu, Xin; Roberts, Michael S

    2017-01-01

    Multiphoton microscopy (MPM) has become increasingly popular and widely used in both basic and clinical liver studies over the past few years. This technology provides insights into deep live tissues with less photobleaching and phototoxicity, which helps us to better understand the cellular morphology, microenvironment, immune responses and spatiotemporal dynamics of drugs and therapeutic cells in the healthy and diseased liver. This review summarizes the principles, opportunities, applications and limitations of MPM in hepatology. A key emphasis is on the use of fluorescence lifetime imaging (FLIM) to add additional quantification and specificity to the detection of endogenous fluorescent species in the liver as well as exogenous molecules and nanoparticles that are applied to the liver in vivo. We anticipate that in the near future MPM-FLIM will advance our understanding of the cellular and molecular mechanisms of liver diseases, and will be evaluated from bench to bedside, leading to real-time histology of human liver diseases. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Label-free detection of breast masses using multiphoton microscopy.

    Directory of Open Access Journals (Sweden)

    Xiufeng Wu

    Full Text Available Histopathology forms the gold standard for the diagnosis of breast cancer. Multiphoton microscopy (MPM has been proposed to be a potentially powerful adjunct to current histopathological techniques. A label-free imaging based on two- photon excited fluorescence and second-harmonic generation is developed for differentiating normal breast tissues, benign, as well as breast cancer tissues. Human breast biopsies (including human normal breast tissues, benign as well as breast cancer tissues that are first imaged (fresh, unfixed, and unstained with MPM and are then processed for routine H-E histopathology. Our results suggest that the MPM images, obtained from these unprocessed biopsies, can readily distinguish between benign lesions and breast cancers. In the tissues of breast cancers, MPM showed that the tumor cells displayed marked cellular and nuclear pleomorphism. The tumor cells, characterized by irregular size and shape, enlarged nuclei, and increased nuclear-cytoplasmic ratio, infiltrated into disrupted connective tissue, leading to the loss of second-harmonic generation signals. For breast cancer, MPM diagnosis was 100% correct because the tissues of breast cancers did not have second-harmonic generation signals in MPM imaging. On the contrary, in benign breast masses, second-harmonic generation signals could be seen easily in MPM imaging. These observations indicate that MPM could be an important potential tool to provide label-free noninvasive diagnostic impressions that can guide surgeon in biopsy and patient management.

  14. Distinguishing human normal or cancerous esophagus tissue ex vivo using multiphoton microscopy

    International Nuclear Information System (INIS)

    Liu, N R; Chen, G N; Wu, S S; Chen, R

    2014-01-01

    Application of multiphoton microscopy (MPM) to clinical cancer research has greatly developed over the last few years. In this paper, we mainly focus on two-photon excitation fluorescence (TPEF) and second harmonic generation (SHG) for investigating esophageal cancer. We chiefly discuss the SHG/TPEF image and spectral characteristics of normal and cancerous esophagus submucosa with the combined multi-channel imaging mode and Lambda mode of a multiphoton microscope (LSM 510 META). Great differences can be detected, such as collagen content and morphology, glandular-shaped cancer cells, TPEF/SHG intensity ratio, and so on, which demonstrate that the multiphoton imaging technique has the potential ability for minimally-invasive early cancer diagnosis. (paper)

  15. Distinguishing human normal or cancerous esophagus tissue ex vivo using multiphoton microscopy

    Science.gov (United States)

    Liu, N. R.; Chen, G. N.; Wu, S. S.; Chen, R.

    2014-02-01

    Application of multiphoton microscopy (MPM) to clinical cancer research has greatly developed over the last few years. In this paper, we mainly focus on two-photon excitation fluorescence (TPEF) and second harmonic generation (SHG) for investigating esophageal cancer. We chiefly discuss the SHG/TPEF image and spectral characteristics of normal and cancerous esophagus submucosa with the combined multi-channel imaging mode and Lambda mode of a multiphoton microscope (LSM 510 META). Great differences can be detected, such as collagen content and morphology, glandular-shaped cancer cells, TPEF/SHG intensity ratio, and so on, which demonstrate that the multiphoton imaging technique has the potential ability for minimally-invasive early cancer diagnosis.

  16. Multiphoton processes in the field of two-frequency circularly polarized plane electromagnetic waves

    International Nuclear Information System (INIS)

    Yu, An

    1997-01-01

    The authors solve Dirac's equation for an electron in the field of a two-frequency plane electromagnetic wave, deriving general formulae for the probabilities of radiation of a photon by the electron, and for the probabilities for pair production by a photon when the two-frequency wave is circularly polarized. In contrast to the case of a monochromatic-plane electromagnetic wave, when an electron is in the field of a two-frequency circularly polarized wave, besides the absorption of multiphotons and emission of simple harmonics of the individual waves, stimulated multiphoton emission processes and various composite harmonic-photon emission processes are occurred: when a high-energy photon is in a such a field, multiphoton processes also follow the pair production processes

  17. Processing multiphoton states through operation on a single photon: Methods and applications

    International Nuclear Information System (INIS)

    Lin Qing; He Bing; Bergou, Janos A.; Ren, Yuhang

    2009-01-01

    Multiphoton states are widely applied in quantum information technology. By the methods presented in this paper, the structure of a multiphoton state in the form of multiple single-photon qubit products can be mapped to a single-photon qudit, which could also be in a separable product with other photons. This makes possible the manipulation of such multiphoton states by processing single-photon states. The optical realization of unknown qubit discrimination [B. He, J. A. Bergou, and Y.-H. Ren, Phys. Rev. A 76, 032301 (2007)] is simplified with the transformation methods. Another application is the construction of quantum logic gates, where the inverse transformations back to the input state spaces are also necessary. We especially show that the modified setups to implement the transformations can realize the deterministic multicontrol gates (including Toffoli gate) operating directly on the products of single-photon qubits.

  18. Imaging rat esophagus using combination of reflectance confocal and multiphoton microscopy

    International Nuclear Information System (INIS)

    Zhuo, S M; Chen, J X; Jiang, X S; Lu, K C; Xie, S S

    2008-01-01

    We combine reflectance confocal microscopy (RCM) with multiphoton microscopy (MPM) to image rat esophagus. The two imaging modalities allow detection of layered–resolved complementary information from esophagus. In the keratinizing layer, the keratinocytes boundaries can be characterized by RCM, while the keratinocytes cytoplasm (keratin) can be further imaged by multiphoton autofluorescence signal. In the epithelium, the epithelial cellular boundaries and nucleus can be detected by RCM, and MPM can be used for imaging epithelial cell cytoplasm and monitoring metabolic state of epithelium. In the stroma, multiphoton autofluorescence signal is used to image elastin and second harmonic generation signal is utilized to detect collagen, while RCM is used to determine the optical property of stroma. Overall, these results suggest that the combination of RCM and MPM has potential to provide more important and comprehensive information for early diagnosis of esophageal cancer

  19. Polarization control of multi-photon absorption under intermediate femtosecond laser field

    International Nuclear Information System (INIS)

    Cheng Wenjing; Liang Guo; Wu Ping; Liu Pei; Jia Tianqing; Sun Zhenrong; Zhang Shian

    2017-01-01

    It has been shown that the femtosecond laser polarization modulation is a very simple and well-established method to control the multi-photon absorption process by the light–matter interaction. Previous studies mainly focused on the multi-photon absorption control in the weak field. In this paper, we further explore the polarization control behavior of multi-photon absorption process in the intermediate femtosecond laser field. In the weak femtosecond laser field, the second-order perturbation theory can well describe the non-resonant two-photon absorption process. However, the higher order nonlinear effect (e.g., four-photon absorption) can occur in the intermediate femtosecond laser field, and thus it is necessary to establish new theoretical model to describe the multi-photon absorption process, which includes the two-photon and four-photon transitions. Here, we construct a fourth-order perturbation theory to study the polarization control behavior of this multi-photon absorption under the intermediate femtosecond laser field excitation, and our theoretical results show that the two-photon and four-photon excitation pathways can induce a coherent interference, while the coherent interference is constructive or destructive that depends on the femtosecond laser center frequency. Moreover, the two-photon and four-photon transitions have the different polarization control efficiency, and the four-photon absorption can obtain the higher polarization control efficiency. Thus, the polarization control efficiency of the whole excitation process can be increased or decreased by properly designing the femtosecond laser field intensity and laser center frequency. These studies can provide a clear physical picture for understanding and controlling the multi-photon absorption process in the intermediate femtosecond laser field, and also can provide a theoretical guidance for the future experimental realization. (paper)

  20. Study on infrared multiphoton excitation of the linear triatomic molecule by the Lie-algebra approach

    International Nuclear Information System (INIS)

    Feng, H.; Zheng, Y.; Ding, S.

    2007-01-01

    Infrared multiphoton vibrational excitation of the linear triatomic molecule has been studied using the quadratic anharmonic Lie-algebra model, unitary transformations, and Magnus approximation. An explicit Lie-algebra expression for the vibrational transition probability is obtained by using a Lie-algebra approach. This explicit Lie-algebra expressions for time-evolution operator and vibrational transition probabilities make the computation clearer and easier. The infrared multiphoton vibrational excitation of the DCN linear tri-atomic molecule is discussed as an example

  1. Additive Manufacture of Three Dimensional Nanocomposite Based Objects through Multiphoton Fabrication

    Directory of Open Access Journals (Sweden)

    Yaan Liu

    2016-09-01

    Full Text Available Three-dimensional structures prepared from a gold-polymer composite formulation have been fabricated using multiphoton lithography. In this process, gold nanoparticles were simultaneously formed through photoreduction whilst polymerisation of two possible monomers was promoted. The monomers, trimethylopropane triacrylate (TMPTA and pentaerythritol triacrylate (PETA were mixed with a gold salt, but it was found that the addition of a ruthenium(II complex enhanced both the geometrical uniformity and integrity of the polymerised/reduced material, enabling the first production of 3D gold-polymer structures by single step multiphoton lithography.

  2. Multiphoton microscopy for the in-situ investigation of cellular processes and integrity in cryopreservation.

    Science.gov (United States)

    Doerr, Daniel; Stark, Martin; Ehrhart, Friederike; Zimmermann, Heiko; Stracke, Frank

    2009-08-01

    In this study we demonstrate a new noninvasive imaging method to monitor freezing processes in biological samples and to investigate life in the frozen state. It combines a laser scanning microscope with a computer-controlled cryostage. Nearinfrared (NIR) femtosecond laser pulses evoke the fluorescence of endogenous fluorophores and fluorescent labels due to multiphoton absorption.The inherent optical nonlinearity of multiphoton absorption allows 3D fluorescence imaging for optical tomography of frozen biological material in-situ. As an example for functional imaging we use fluorescence lifetime imaging (FLIM) to create images with chemical and physical contrast.

  3. High-resolution multiphoton microscopy with a low-power continuous wave laser pump.

    Science.gov (United States)

    Chen, Xiang-Dong; Li, Shen; Du, Bo; Dong, Yang; Wang, Ze-Hao; Guo, Guang-Can; Sun, Fang-Wen

    2018-02-15

    Multiphoton microscopy (MPM) has been widely used for three-dimensional biological imaging. Here, based on the photon-induced charge state conversion process, we demonstrated a low-power high-resolution MPM with a nitrogen vacancy (NV) center in diamond. Continuous wave green and orange lasers were used to pump and detect the two-photon charge state conversion, respectively. The power of the laser for multiphoton excitation was 40 μW. Both the axial and lateral resolutions were improved approximately 1.5 times compared with confocal microscopy. The results can be used to improve the resolution of the NV center-based quantum sensing and biological imaging.

  4. Complementary roles of platelets and coagulation in thrombus formation on plaques acutely ruptured by targeted ultrasound treatment: a novel intravital model.

    Science.gov (United States)

    Kuijpers, M J E; Gilio, K; Reitsma, S; Nergiz-Unal, R; Prinzen, L; Heeneman, S; Lutgens, E; van Zandvoort, M A M J; Nieswandt, B; Egbrink, M G A Oude; Heemskerk, J W M

    2009-01-01

    Atherothrombosis is a major cause of cardiovascular events. However, animal models to study this process are scarce. We describe the first murine model of acute thrombus formation upon plaque rupture to study atherothrombosis by intravital fluorescence microscopy. Localized rupture of an atherosclerotic plaque in a carotid artery from Apoe(-/-) mice was induced in vivo using ultrasound. Rupture of the plaque and formation of localized thrombi were verified by two-photon laser scanning microscopy (TPLSM) in isolated arteries, and by immunohistochemistry. The thrombotic reaction was quantified by intravital fluorescence microscopy. Inspection of the ultrasound-treated plaques by histochemistry and TPLSM demonstrated local damage, collagen exposure, luminal thrombus formation as well as intra-plaque intrusion of erythrocytes and fibrin. Ultrasound treatment of healthy carotid arteries resulted in endothelial damage and limited platelet adhesion. Real-time intravital fluorescence microscopy demonstrated rapid platelet deposition on plaques and formation of a single thrombus that remained subocclusive. The thrombotic process was antagonized by thrombin inhibition, or by blocking of collagen or adenosine diphosphate receptor pathways. Multiple thrombi were formed in 70% of mice lacking CD40L. Targeted rupture of murine plaques results in collagen exposure and non-occlusive thrombus formation. The thrombotic process relies on platelet activation as well as on thrombin generation and coagulation, and is sensitive to established and novel antithrombotic medication. This model provides new possibilities to study atherothrombosis in vivo.

  5. The Use of Intravital Two-Photon and Thick Section Confocal Imaging to Analyze B Lymphocyte Trafficking in Lymph Nodes and Spleen.

    Science.gov (United States)

    Park, Chung; Hwang, Il-Young; Kehrl, John H

    2018-01-01

    Intravital two-photon laser scanning microscopy (TP-LSM) has allowed the direct observation of immune cells in intact organs of living animals. In the B cell biology field TP-LSM has detailed the movement of B cells in high endothelial venules and during their transmigration into lymph organs; described the movement and positioning of B cells within lymphoid organs; outlined the mechanisms by which antigen is delivered to B cells; observed B cell interacting with T cells, other cell types, and even with pathogens; and delineated the egress of B cells from the lymph node (LN) parenchyma into the efferent lymphatics. As the quality of TP-LSM improves and as new fluorescent probes become available additional insights into B cell behavior and function await new investigations. Yet intravital TP-LSM has some disadvantages including a lower resolution than standard confocal microscopy, a narrow imaging window, and a shallow depth of imaging. We have found that supplementing intravital TP-LSM with conventional confocal microscopy using thick LN sections helps to overcome some of these shortcomings. Here, we describe procedures for visualizing the behavior and trafficking of fluorescently labeled, adoptively transferred antigen-activated B cells within the inguinal LN of live mice using two-photon microscopy. Also, we introduce procedures for fixed thick section imaging using standard confocal microscopy, which allows imaging of fluorescently labeled cells deep in the LN cortex and in the spleen with high resolution.

  6. Multi-Photon Absorption Spectra: A Comparison Between Transmittance Change and Fluorescence Methods

    Science.gov (United States)

    2015-05-21

    AFRL-OSR-VA-TR-2015-0134 multi-photon absorption spectra Cleber Mendonca INSTITUTO DE FISICA DE SAO CARLOS Final Report 05/21/2015 DISTRIBUTION A...5f. WORK UNIT NUMBER 7. PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES) Instituto de Fisica de Sao Carlos - Universidade de Sao Paulo Av

  7. The layered-resolved microstructure and spectroscopy of mouse oral mucosa using multiphoton microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Zhuo Shuangmu [Key Laboratory of Optoelectronic Science and Technology for Medicine, Fujian Normal University, Ministry of Education, Fuzhou 350007 (China); Chen Jianxin [Key Laboratory of Optoelectronic Science and Technology for Medicine, Fujian Normal University, Ministry of Education, Fuzhou 350007 (China); Jiang Xingshan [Key Laboratory of Optoelectronic Science and Technology for Medicine, Fujian Normal University, Ministry of Education, Fuzhou 350007 (China); Xie Shusen [Key Laboratory of Optoelectronic Science and Technology for Medicine, Fujian Normal University, Ministry of Education, Fuzhou 350007 (China); Chen Rong [Key Laboratory of Optoelectronic Science and Technology for Medicine, Fujian Normal University, Ministry of Education, Fuzhou 350007 (China); Cao Ning [Fujian Medical University, Fuzhou 350004 (China); Zou Qilian [Fujian Medical University, Fuzhou 350004 (China); Xiong Shuyuan [Fujian Medical University, Fuzhou 350004 (China)

    2007-08-21

    The layered-resolved microstructure and spectroscopy of mouse oral mucosa are obtained using a combination of multiphoton imaging and spectral analysis with different excitation wavelengths. In the keratinizing layer, the keratinocytes microstructure can be characterized and the keratinizing thickness can be measured. The keratin fluorescence signal can be further characterized by emission maxima at 510 nm. In the epithelium, the cellular microstructure can be quantitatively visualized with depth and the epithelium thickness can be determined by multiphoton imaging excited at 730 nm. The study also shows that the epithelial spectra excited at 810 nm, showing a combination of NADH and FAD fluorescence, can be used for the estimation of the metabolic state in epithelium. Interestingly, a second-harmonic generation (SHG) signal from DNA was observed for the first time within the epithelial layer in backscattering geometry and provides the possibility of analyzing the chromatin structure. In the stroma, the combination of multiphoton imaging and spectral analysis excited at 850 nm in tandem can obtain quantitative information regarding the biomorphology and biochemistry of stroma. Specifically, the microstructure of collagen, minor salivary glands and elastic fibers, and the optical property of the stroma can be quantitatively displayed. Overall, these results suggest that the combination of multiphoton imaging and spectral analysis with different excitation wavelengths has the potential to provide important and comprehensive information for early diagnosis of oral cancer.

  8. Statistical properties of multiphoton time-dependent three-boson coupled oscillators

    Czech Academy of Sciences Publication Activity Database

    Abdalla, M. S.; Peřina, Jan; Křepelka, Jaromír

    2006-01-01

    Roč. 23, č. 6 (2006), s. 1146-1160 ISSN 0740-3224 R&D Projects: GA MŠk(CZ) OC P11.003 Institutional research plan: CEZ:AV0Z10100522 Keywords : quantum statistic * coupled oscillators * multiphoton Subject RIV: BH - Optics, Masers, Lasers Impact factor: 2.002, year: 2006

  9. Temporal shaping of nanosecond CO2 laser pulses in multiphoton saturable absorbers

    International Nuclear Information System (INIS)

    Haglund, R.F. Jr.

    1981-01-01

    It was shown that substantial temporal distortion of nanosecond 10.6 μm laser pulses occurs in traversing multiphoton saturable absorbers. The risetime and pulse delay effects appear to depend both on fluence and wavelength, and to be qualitatively consistent with predictions of a simple two-level absorption model

  10. Electron-Nuclear Energy Sharing in Above-Threshold Multiphoton Dissociative Ionization of H2

    DEFF Research Database (Denmark)

    Wu, J.; Kunitski, M.; Pitzer, M.

    2013-01-01

    We report experimental observation of the energy sharing between electron and nuclei in above-threshold multiphoton dissociative ionization of H2 by strong laser fields. The absorbed photon energy is shared between the ejected electron and nuclei in a correlated fashion, resulting in multiple...... diagonal lines in their joint energy spectrum governed by the energy conservation of all fragment particles....

  11. Multiphoton absorption probabilities in strong laser fields with application to H-

    International Nuclear Information System (INIS)

    Mu, X.; University of Oregon, Eugene, OR

    1990-01-01

    The commonly used Keldysh multiphoton ionization rate is shown to follow from the zeroth-order approximation of an exact expression, based on the formal time-independent theory of scattering. The formulation is applied to the loosely bound H - system; good agreement is obtained with a recent experimental measurement

  12. Self-referenced axial chromatic dispersion measurement in multiphoton microscopy through 2-color THG imaging.

    Science.gov (United States)

    Du, Yu; Zhuang, Ziwei; He, Jiexing; Liu, Hongji; Qiu, Ping; Wang, Ke

    2018-05-16

    With tunable excitation light, multiphoton microscopy (MPM) is widely used for imaging biological structures at subcellular resolution. Axial chromatic dispersion, present in virtually every transmissive optical system including the multiphoton microscope, leads to focal (and the resultant image) plane separation. Here we demonstrate experimentally a technique to measure the axial chromatic dispersion in a multiphoton microscope, using simultaneous 2-color third-harmonic generation (THG) imaging excited by a 2-color soliton source with tunable wavelength separation. Our technique is self-referenced, eliminating potential measurement error when 1-color tunable excitation light is used which necessitates reciprocating motion of the mechanical translation stage. Using this technique, we demonstrate measured axial chromatic dispersion with 2 different objective lenses in a multiphoton microscope. Further measurement in a biological sample also indicates that this axial chromatic dispersion, in combination with 2-color imaging, may open up opportunity for simultaneous imaging of two different axial planes. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  13. Exact results for emission from one and two atoms in an ideal cavity at multiphoton resonance

    International Nuclear Information System (INIS)

    Fam Le Kien; Shumovskij, A.S.; Tran Quang.

    1987-01-01

    The emission from the system of one or two two-level atoms in an ideal cavity with one mode at mutiphoton resonance is examined. Exact results for the two-time dipole correlation function and the time-dependent spectra of multiphoton-induced fluorescence are presented

  14. Electron energy spectrum and maximum disruption angle under multi-photon beamstrahlung

    Energy Technology Data Exchange (ETDEWEB)

    Yokoya, Kaoru; Chen, Pisin

    1989-03-01

    The final electron energy spectrum under multi-photon beamstrahlung process is derived analytically in the classical and the intermediate regimes. The maximum disruption angle from the low energy tail of the spectrum is also estimated. The results are then applied to the TLC and the CLIC parameters. 6 refs., 1 fig., 1 tab.

  15. Potential of ultraviolet widefield imaging and multiphoton microscopy for analysis of dehydroergosterol in cellular membranes

    DEFF Research Database (Denmark)

    Wüstner, Daniel; Brewer, Jonathan R.; Bagatolli, Luis

    2011-01-01

    Dehydroergosterol (DHE) is an intrinsically fluorescent sterol with absorption/emission in the ultraviolet (UV) region and biophysical properties similar to those of cholesterol. We compared the potential of UV-sensitive low-light-level wide-field (UV-WF) imaging with that of multiphoton (MP) exc...

  16. Semiclassical analysis of long-wavelength multiphoton processes: The Rydberg atom

    International Nuclear Information System (INIS)

    Vela-Arevalo, Luz V.; Fox, Ronald F.

    2004-01-01

    We study the problem of multiphoton processes for intense, long-wavelength irradiation of atomic and molecular electrons. An exact, nonperturbative approach is applied to the standard vector potential coupling Hamiltonian for a three-dimensional hydrogenlike atom in a microwave field treated semiclassically. Multiphoton probability exchange is calculated in both the velocity and the length gauges, by applying the Goeppert-Mayer gauge transformation. The expansion of the time-dependent solution in terms of Floquet states delineates the mechanism of multiphoton transitions. A detailed analysis of the Floquet states and quasienergies as functions of the field parameters allows us to describe the relation between avoided quasienergy crossings and multiphoton probability exchange. We formulate analytical expressions for the variation of quasienergies and Floquet states with respect to the field parameters, and demonstrate that avoided quasienergy crossings are accompanied by dramatic changes in the Floquet states. Analysis of the Floquet states, for small values of the field strength, yields selection rules for the avoided quasienergy crossings. In the case of strong fields, the simultaneous choice of frequency and strength of the field producing an avoided crossing results in improved ionization probability

  17. Investigations of multiphoton excitation and ionization in a short range potential

    International Nuclear Information System (INIS)

    Susskind, S.M.; Cowley, S.C.; Valeo, E.J.

    1989-02-01

    We introduce an approach to the study of excitation and ionization for a system with a short range potential. In particular, analytical and numerical results are presented for the multiphoton ionization rate, under strong field conditions, of an electron confined by a δ-function potential. 9 refs., 3 figs

  18. Investigations of multiphoton excitation and ionization in a short range potential

    Energy Technology Data Exchange (ETDEWEB)

    Susskind, S.M.; Cowley, S.C.; Valeo, E.J.

    1989-02-01

    We introduce an approach to the study of excitation and ionization for a system with a short range potential. In particular, analytical and numerical results are presented for the multiphoton ionization rate, under strong field conditions, of an electron confined by a delta-function potential. 9 refs., 3 figs.

  19. Improving spatial resolution in quantum imaging beyond the Rayleigh diffraction limit using multiphoton W entangled states

    Energy Technology Data Exchange (ETDEWEB)

    Wen Jianming, E-mail: jianming.wen@gmail.co [National Laboratory of Solid State Microstructures and Department of Physics, Nanjing University, Nanjing 210093 (China); Department of Physics, University of Arkansas, Fayetteville, AR 72701 (United States); Du, Shengwang [Department of Physics, Hong Kong University of Science and Technology, Clear Bay (Hong Kong); Xiao Min [National Laboratory of Solid State Microstructures and Department of Physics, Nanjing University, Nanjing 210093 (China); Department of Physics, University of Arkansas, Fayetteville, AR 72701 (United States); School of Modern Engineering and Applied Science, Nanjing University, Nanjing 210093 (China)

    2010-08-23

    Using multiphoton entangled states, we demonstrate improving spatial imaging resolution beyond the Rayleigh diffraction limit in the quantum imaging process. In particular, we examine resolution enhancement using triphoton W state and a factor of 2 is achievable as with the use of the Greenberger-Horne-Zeilinger state, compared to using a classical-light source.

  20. Generating multiphoton Greenberger-Horne-Zeilinger states with weak cross-Kerr nonlinearity

    International Nuclear Information System (INIS)

    Jin, Guang-Sheng; Lin, Yuan; Wu, Biao

    2007-01-01

    We propose a scheme to generate polarization-entangled multiphoton Greenberger-Horne-Zeilinger states with weak cross-Kerr nonlinearity based on controlled bus rotation and subsequent homodyne measurement. Our method is simple in operation and has high success probabilities with near perfect fidelities in an ideal case

  1. Multi-photon entanglement and applications in quantum information

    Energy Technology Data Exchange (ETDEWEB)

    Schmid, Christian I.T.

    2008-05-30

    In this thesis, two new linear optics networks are introduced and their application for several quantum information tasks is presented. Spontaneous parametric down conversion, is used in different configurations to provide the input states for the networks. The first network is a new design of a controlled phase gate which is particularly interesting for applications in multi-photon experiments as it constitutes an improvement of former realizations with respect to stability and reliability. This is explicitly demonstrated by employing the gate in four-photon experiments. In this context, a teleportation and entanglement swapping protocol is performed in which all four Bell states are distinguished by means of the phase gate. A similar type of measurement applied to the subsystem parts of two copies of a quantum state, allows further the direct estimation of the state's entanglement in terms of its concurrence. Finally, starting from two Bell states, the controlled phase gate is applied for the observation of a four photon cluster state. The analysis of the results focuses on measurement based quantum computation, the main usage of cluster states. The second network, fed with the second order emission of non-collinear type ii spontaneous parametric down conversion, constitutes a tunable source of a whole family of states. Up to now the observation of one particular state required one individually tailored setup. With the network introduced here many different states can be obtained within the same arrangement by tuning a single, easily accessible experimental parameter. These states exhibit many useful properties and play a central role in several applications of quantum information. Here, they are used for the solution of a four-player quantum Minority game. It is shown that, by employing four-qubit entanglement, the quantum version of the game clearly outperforms its classical counterpart. Experimental data obtained with both networks are utilized to

  2. Multi-photon entanglement and applications in quantum information

    International Nuclear Information System (INIS)

    Schmid, Christian I.T.

    2008-01-01

    In this thesis, two new linear optics networks are introduced and their application for several quantum information tasks is presented. Spontaneous parametric down conversion, is used in different configurations to provide the input states for the networks. The first network is a new design of a controlled phase gate which is particularly interesting for applications in multi-photon experiments as it constitutes an improvement of former realizations with respect to stability and reliability. This is explicitly demonstrated by employing the gate in four-photon experiments. In this context, a teleportation and entanglement swapping protocol is performed in which all four Bell states are distinguished by means of the phase gate. A similar type of measurement applied to the subsystem parts of two copies of a quantum state, allows further the direct estimation of the state's entanglement in terms of its concurrence. Finally, starting from two Bell states, the controlled phase gate is applied for the observation of a four photon cluster state. The analysis of the results focuses on measurement based quantum computation, the main usage of cluster states. The second network, fed with the second order emission of non-collinear type ii spontaneous parametric down conversion, constitutes a tunable source of a whole family of states. Up to now the observation of one particular state required one individually tailored setup. With the network introduced here many different states can be obtained within the same arrangement by tuning a single, easily accessible experimental parameter. These states exhibit many useful properties and play a central role in several applications of quantum information. Here, they are used for the solution of a four-player quantum Minority game. It is shown that, by employing four-qubit entanglement, the quantum version of the game clearly outperforms its classical counterpart. Experimental data obtained with both networks are utilized to demonstrate

  3. Computational code in atomic and nuclear quantum optics: Advanced computing multiphoton resonance parameters for atoms in a strong laser field

    Science.gov (United States)

    Glushkov, A. V.; Gurskaya, M. Yu; Ignatenko, A. V.; Smirnov, A. V.; Serga, I. N.; Svinarenko, A. A.; Ternovsky, E. V.

    2017-10-01

    The consistent relativistic energy approach to the finite Fermi-systems (atoms and nuclei) in a strong realistic laser field is presented and applied to computing the multiphoton resonances parameters in some atoms and nuclei. The approach is based on the Gell-Mann and Low S-matrix formalism, multiphoton resonance lines moments technique and advanced Ivanov-Ivanova algorithm of calculating the Green’s function of the Dirac equation. The data for multiphoton resonance width and shift for the Cs atom and the 57Fe nucleus in dependence upon the laser intensity are listed.

  4. Data-adaptive image-denoising for detecting and quantifying nanoparticle entry in mucosal tissues through intravital 2-photon microscopy

    Directory of Open Access Journals (Sweden)

    Torsten Bölke

    2014-11-01

    Full Text Available Intravital 2-photon microscopy of mucosal membranes across which nanoparticles enter the organism typically generates noisy images. Because the noise results from the random statistics of only very few photons detected per pixel, it cannot be avoided by technical means. Fluorescent nanoparticles contained in the tissue may be represented by a few bright pixels which closely resemble the noise structure. We here present a data-adaptive method for digital denoising of datasets obtained by 2-photon microscopy. The algorithm exploits both local and non-local redundancy of the underlying ground-truth signal to reduce noise. Our approach automatically adapts the strength of noise suppression in a data-adaptive way by using a Bayesian network. The results show that the specific adaption to both signal and noise characteristics improves the preservation of fine structures such as nanoparticles while less artefacts were produced as compared to reference algorithms. Our method is applicable to other imaging modalities as well, provided the specific noise characteristics are known and taken into account.

  5. Differentiating the two main histologic categories of fibroadenoma tissue from normal breast tissue by using multiphoton microscopy.

    Science.gov (United States)

    Nie, Y T; Wu, Y; Fu, F M; Lian, Y E; Zhuo, S M; Wang, C; Chen, J X

    2015-04-01

    Multiphoton microscopy has become a novel biological imaging technique that allows cellular and subcellular microstructure imaging based on two-photon excited fluorescence and second harmonic generation. In this work, we used multiphoton microscopy to obtain the high-contrast images of human normal breast tissue and two main histologic types of fibroadenoma (intracanalicular, pericanalicular). Moreover, quantitative image analysis was performed to characterize the changes of collagen morphology (collagen content, collagen orientation). The results show that multiphoton microscopy combined with quantitative method has the ability to identify the characteristics of fibroadenoma including changes of the duct architecture and collagen morphology in stroma. With the advancement of multiphoton microscopy, we believe that the technique has great potential to be a real-time histopathological diagnostic tool for intraoperative detection of fibroadenoma in the future. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

  6. In vivo real-time multiphoton imaging of T lymphocytes in the mouse brain after experimental stroke

    DEFF Research Database (Denmark)

    Fumagalli, Stefano; Coles, Jonathan A; Ejlerskov, Patrick

    2011-01-01

    To gain a better understanding of T cell behavior after stroke, we have developed real-time in vivo brain imaging of T cells by multiphoton microscopy after middle cerebral artery occlusion.......To gain a better understanding of T cell behavior after stroke, we have developed real-time in vivo brain imaging of T cells by multiphoton microscopy after middle cerebral artery occlusion....

  7. In vivo 3D measurement of moxifloxacin and gatifloxacin distributions in the mouse cornea using multiphoton microscopy

    OpenAIRE

    Lee, Seunghun; Lee, Jun Ho; Park, Jin Hyoung; Yoon, Yeoreum; Chung, Wan Kyun; Tchah, Hungwon; Kim, Myoung Joon; Kim, Ki Hean

    2016-01-01

    Moxifloxacin and gatifloxacin are fourth-generation fluoroquinolone antibiotics used in the clinic to prevent or treat ocular infections. Their pharmacokinetics in the cornea is usually measured from extracted ocular fluids or tissues, and in vivo direct measurement is difficult. In this study multiphoton microscopy (MPM), which is a 3D optical microscopic technique based on multiphoton fluorescence, was applied to the measurement of moxifloxacin and gatifloxacin distribution in the cornea. I...

  8. The selective Cox-2 inhibitor Celecoxib suppresses angiogenesis and growth of secondary bone tumors: An intravital microscopy study in mice

    International Nuclear Information System (INIS)

    Klenke, Frank Michael; Gebhard, Martha-Maria; Ewerbeck, Volker; Abdollahi, Amir; Huber, Peter E; Sckell, Axel

    2006-01-01

    The inhibition of angiogenesis is a promising strategy for the treatment of malignant primary and secondary tumors in addition to established therapies such as surgery, chemotherapy, and radiation. There is strong experimental evidence in primary tumors that Cyclooxygenase-2 (Cox-2) inhibition is a potent mechanism to reduce angiogenesis. For bone metastases which occur in up to 85% of the most frequent malignant primary tumors, the effects of Cox-2 inhibition on angiogenesis and tumor growth remain still unclear. Therefore, the aim of this study was to investigate the effects of Celecoxib, a selective Cox-2 inhibitor, on angiogenesis, microcirculation and growth of secondary bone tumors. In 10 male severe combined immunodeficient (SCID) mice, pieces of A549 lung carcinomas were implanted into a newly developed cranial window preparation where the calvaria serves as the site for orthotopic implantation of the tumors. From day 8 after tumor implantation, five animals (Celecoxib) were treated daily with Celecoxib (30 mg/kg body weight, s.c.), and five animals (Control) with the equivalent amount of the CMC-based vehicle. Angiogenesis, microcirculation, and growth of A549 tumors were analyzed by means of intravital microscopy. Apoptosis was quantified using the TUNEL assay. Treatment with Celecoxib reduced both microvessel density and tumor growth. TUNEL reaction showed an increase in apoptotic cell death of tumor cells after treatment with Celecoxib as compared to Controls. Celecoxib is a potent inhibitor of tumor growth of secondary bone tumors in vivo which can be explained by its anti-angiogenic and pro-apoptotic effects. The results indicate that a combination of established therapy regimes with Cox-2 inhibition represents a possible application for the treatment of bone metastases

  9. Direct visualization of electroporation-assisted in vivo gene delivery to tumors using intravital microscopy – spatial and time dependent distribution

    Directory of Open Access Journals (Sweden)

    Dachs Gabi U

    2004-11-01

    Full Text Available Abstract Background Electroporation is currently receiving much attention as a way to increase drug and DNA delivery. Recent studies demonstrated the feasibility of electrogene therapy using a range of therapeutic genes for the treatment of experimental tumors. However, the transfection efficiency of electroporation-assisted DNA delivery is still low compared to viral methods and there is a clear need to optimize this approach. In order to optimize treatment, knowledge about spatial and time dependency of gene expression following delivery is of utmost importance in order to improve gene delivery. Intravital microscopy of tumors growing in dorsal skin fold window chambers is a useful method for monitoring gene transfection, since it allows non-invasive dynamic monitoring of gene expression in tumors in a live animal. Methods Intravital microscopy was used to monitor real time spatial distribution of the green fluorescent protein (GFP and time dependence of transfection efficiency in syngeneic P22 rat tumor model. DNA alone, liposome-DNA complexes and electroporation-assisted DNA delivery using two different sets of electric pulse parameters were compared. Results Electroporation-assisted DNA delivery using 8 pulses, 600 V/cm, 5 ms, 1 Hz was superior to other methods and resulted in 22% increase in fluorescence intensity in the tumors up to 6 days post-transfection, compared to the non-transfected area in granulation tissue. Functional GFP was detected within 5 h after transfection. Cells expressing GFP were detected throughout the tumor, but not in the surrounding tissue that was not exposed to electric pulses. Conclusions Intravital microscopy was demonstrated to be a suitable method for monitoring time and spatial distribution of gene expression in experimental tumors and provided evidence that electroporation-assisted gene delivery using 8 pulses, 600 V/cm, 5 ms, 1 Hz is an effective method, resulting in early onset and homogenous

  10. Direct visualization of electroporation-assisted in vivo gene delivery to tumors using intravital microscopy – spatial and time dependent distribution

    International Nuclear Information System (INIS)

    Cemazar, Maja; Wilson, Ian; Dachs, Gabi U; Tozer, Gillian M; Sersa, Gregor

    2004-01-01

    Electroporation is currently receiving much attention as a way to increase drug and DNA delivery. Recent studies demonstrated the feasibility of electrogene therapy using a range of therapeutic genes for the treatment of experimental tumors. However, the transfection efficiency of electroporation-assisted DNA delivery is still low compared to viral methods and there is a clear need to optimize this approach. In order to optimize treatment, knowledge about spatial and time dependency of gene expression following delivery is of utmost importance in order to improve gene delivery. Intravital microscopy of tumors growing in dorsal skin fold window chambers is a useful method for monitoring gene transfection, since it allows non-invasive dynamic monitoring of gene expression in tumors in a live animal. Intravital microscopy was used to monitor real time spatial distribution of the green fluorescent protein (GFP) and time dependence of transfection efficiency in syngeneic P22 rat tumor model. DNA alone, liposome-DNA complexes and electroporation-assisted DNA delivery using two different sets of electric pulse parameters were compared. Electroporation-assisted DNA delivery using 8 pulses, 600 V/cm, 5 ms, 1 Hz was superior to other methods and resulted in 22% increase in fluorescence intensity in the tumors up to 6 days post-transfection, compared to the non-transfected area in granulation tissue. Functional GFP was detected within 5 h after transfection. Cells expressing GFP were detected throughout the tumor, but not in the surrounding tissue that was not exposed to electric pulses. Intravital microscopy was demonstrated to be a suitable method for monitoring time and spatial distribution of gene expression in experimental tumors and provided evidence that electroporation-assisted gene delivery using 8 pulses, 600 V/cm, 5 ms, 1 Hz is an effective method, resulting in early onset and homogenous distribution of gene expression in the syngeneic P22 rat tumor model

  11. Development of novel murine mammary imaging windows to examine wound healing effects on leukocyte trafficking in mammary tumors with intravital imaging.

    Science.gov (United States)

    Sobolik, Tammy; Su, Ying-Jun; Ashby, Will; Schaffer, David K; Wells, Sam; Wikswo, John P; Zijlstra, Andries; Richmond, Ann

    2016-01-01

    We developed mammary imaging windows (MIWs) to evaluate leukocyte infiltration and cancer cell dissemination in mouse mammary tumors imaged by confocal microscopy. Previous techniques relied on surgical resection of a skin flap to image the tumor microenvironment restricting imaging time to a few hours. Utilization of mammary imaging windows offers extension of intravital imaging of the tumor microenvironment. We have characterized strengths and identified some previously undescribed potential weaknesses of MIW techniques. Through iterative enhancements of a transdermal portal we defined conditions for improved quality and extended confocal imaging time for imaging key cell-cell interactions in the tumor microenvironment.

  12. Multiphoton atomic ionization in the field of a very short laser pulse

    International Nuclear Information System (INIS)

    Popov, V.S.

    2001-01-01

    Closed analytic expressions are derived for the probability of multiphoton atomic and ionic ionization in a variable electric field E(t), which are applicable for arbitrary Keldysh parameters γ. Dependencies of the ionization probability and photoelectron pulse spectrum on the shape of a very short laser pulse are analyzed. Examples of pulse fields of various forms, including a modulated light pulse with a Gaussian or Lorentz envelope, are considered in detail. The interference effect in the photoelectron energy spectrum during atomic ionization by a periodic field of a general form is examined. The range of applicability of the adiabatic approximation in the multiphoton ionization theory is discussed. The imaginary time method is used in the calculations, which allows the probability of particle tunneling through oscillating barriers to be effectively calculated

  13. Dirac Equation in (1 +1 )-Dimensional Curved Spacetime and the Multiphoton Quantum Rabi Model

    Science.gov (United States)

    Pedernales, J. S.; Beau, M.; Pittman, S. M.; Egusquiza, I. L.; Lamata, L.; Solano, E.; del Campo, A.

    2018-04-01

    We introduce an exact mapping between the Dirac equation in (1 +1 )-dimensional curved spacetime (DCS) and a multiphoton quantum Rabi model (QRM). A background of a (1 +1 )-dimensional black hole requires a QRM with one- and two-photon terms that can be implemented in a trapped ion for the quantum simulation of Dirac particles in curved spacetime. We illustrate our proposal with a numerical analysis of the free fall of a Dirac particle into a (1 +1 )-dimensional black hole, and find that the Zitterbewegung effect, measurable via the oscillatory trajectory of the Dirac particle, persists in the presence of gravity. From the duality between the squeezing term in the multiphoton QRM and the metric coupling in the DCS, we show that gravity generates squeezing of the Dirac particle wave function.

  14. Label-free identification of intestinal metaplasia in the stomach using multiphoton microscopy

    International Nuclear Information System (INIS)

    Wu, G; Wei, J; Zheng, Z; Ye, J; Zeng, S

    2014-01-01

    The early diagnosis of intestinal metaplasia (IM) in the stomach together with effective therapeutic interventions is crucial to reducing the mortality-rates of the patients associated with gastric cancer. However, it is challenging during conventional white-light endoscopy, and histological analysis remains the ‘gold standard’ for the final diagnosis. Here, we describe a label-free imaging method, multiphoton microscopy (MPM), for the identification of IM in the stomach. It was found that multiphoton imaging provides cellular and subcellular details to the identification of IM from normal gastric tissues. In particular, there is significant difference in the population density of goblet cells between normal and IM gastric tissues, providing substantial potential to become a quantitative intrinsic marker for in vivo clinical diagnosis of early gastric lesions. To our knowledge, this is the first demonstration of the potential of MPM for the identification of IM. (letters)

  15. Identification of normal and cancerous human colorectal muscularis propria by multiphoton microscopy in different sections

    International Nuclear Information System (INIS)

    Zhou, Yi; Li, Lianhuang; Zhuo, Shuangmu; Zhu, Xiaoqin; Chen, Jianxin; Chen, Zhifen; Guan, Guoxian; Kang, Deyong

    2016-01-01

    Multiphoton microscopy (MPM) based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) as a potential diagnostic tool is attractive. MPM can effectively provide information about morphological and biochemical changes in biological tissues at the molecular level. In this paper, we attempt to identify normal and cancerous human colorectal muscularis propria by multiphoton microscopy in different sections (both in transverse and longitudinal sections). The results show that MPM can display different microstructure changes in the transverse and longitudinal sections of colorectal muscularis propria. MPM also can quantitatively describe the alteration of collagen content between normal and cancerous muscle layers. These are important pathological findings that MPM images can bring more detailed complementary information about tissue architecture and cell morphology through observing the transverse and longitudinal sections of colorectal muscularis propria. This work demonstrates that MPM can be better for identifying the microstructural characteristics of normal and cancerous human colorectal muscularis propria in different sections. (paper)

  16. Multiphoton ionization of the hydrogen atom by a circularly polarized electromagnetic field

    International Nuclear Information System (INIS)

    Prepelitsa, O.B.

    1999-01-01

    This paper examines the multiphoton ionization of the ground state of the hydrogen atom in the field of a circularly polarized intense electromagnetic wave. To describe the states of photoelectrons, quasiclassical wave functions are introduced that partially allow for the effect of an intense electromagnetic wave and that of the Coulomb potential. Expressions are derived for the angular and energy distributions of photoelectrons with energies much lower than the ionization potential of an unperturbed atom. It is found that, due to allowance for the Coulomb potential in the wave function of the final electron states, the transition probability near the ionization threshold tends to a finite value. In addition, the well-known selection rules for multiphoton transitions in a circularly polarized electromagnetic field are derived in a natural way. Finally, the results are compared with those obtained in the Keldysh-Faisal-Reiss approximation

  17. Multi-photon processes brought about by a laser; Processus multiphotoniques provoques par un laser

    Energy Technology Data Exchange (ETDEWEB)

    Nelson, P [Commissariat a l' Energie Atomique, Saclay (France). Centre d' Etudes Nucleaires

    1965-07-01

    We calculate the critical intensity characterizing the multiphoton processes. The multiphoton effects corresponding to the Compton scattering, the Bremsstrahlung, the photoelectric effect are investigated. The cross sections are evaluated. We show how the introduction of a refractive index, in clothing the photons, allows the elimination of the infrared divergence. The theory seems consistent with experiment. (author) [French] Nous calculons l'intensite critique caracterisant les processus multiphotoniques. Les effets multiphotoniques correspondant a la diffusion Compton, au bremsstrahlung, a l'effet photoelectrique sont etudies. Les sections efficaces sont evaluees. Nous montrons comment l'introduction d'un indice de refraction, en habillant les photons, permet d'eliminer les divergences infra-rouges. La theorie semble compatible avec l'experience. (auteur)

  18. Protocol for generating multiphoton entangled states from quantum dots in the presence of nuclear spin fluctuations

    DEFF Research Database (Denmark)

    Denning, Emil Vosmar; Iles-Smith, Jake; McCutcheon, Dara P. S.

    2017-01-01

    Multiphoton entangled states are a crucial resource for many applications inquantum information science. Semiconductor quantum dots offer a promising route to generate such states by mediating photon-photon correlations via a confinedelectron spin, but dephasing caused by the host nuclear spin...... environment typically limits coherence (and hence entanglement) between photons to the spin T2* time of a few nanoseconds. We propose a protocol for the deterministic generation of multiphoton entangled states that is inherently robust against the dominating slow nuclear spin environment fluctuations, meaning...... that coherence and entanglement is instead limited only by the much longer spin T2 time of microseconds. Unlike previous protocols, the present schemeallows for the generation of very low error probability polarisation encoded three-photon GHZ states and larger entangled states, without the need for spin echo...

  19. Multiphoton Laser Microscopy and Fluorescence Lifetime Imaging for the Evaluation of the Skin

    Directory of Open Access Journals (Sweden)

    Stefania Seidenari

    2012-01-01

    Full Text Available Multiphoton laser microscopy is a new, non-invasive technique providing access to the skin at a cellular and subcellular level, which is based both on autofluorescence and fluorescence lifetime imaging. Whereas the former considers fluorescence intensity emitted by epidermal and dermal fluorophores and by the extra-cellular matrix, fluorescence lifetime imaging (FLIM, is generated by the fluorescence decay rate. This innovative technique can be applied to the study of living skin, cell cultures and ex vivo samples. Although still limited to the clinical research field, the development of multiphoton laser microscopy is thought to become suitable for a practical application in the next few years: in this paper, we performed an accurate review of the studies published so far, considering the possible fields of application of this imaging method and providing high quality images acquired in the Department of Dermatology of the University of Modena.

  20. vuv fluorescence from selective high-order multiphoton excitation of N2

    International Nuclear Information System (INIS)

    Coffee, Ryan N.; Gibson, George N.

    2004-01-01

    Recent fluorescence studies suggest that ultrashort pulse laser excitation may be highly selective. Selective high-intensity laser excitation holds important consequences for the physics of multiphoton processes. To establish the extent of this selectivity, we performed a detailed comparative study of the vacuum ultraviolet fluorescence resulting from the interaction of N 2 and Ar with high-intensity infrared ultrashort laser pulses. Both N 2 and Ar reveal two classes of transitions, inner-valence ns ' l ' . From their pressure dependence, we associate each transition with either plasma or direct laser excitation. Furthermore, we qualitatively confirm such associations with the time dependence of the fluorescence signal. Remarkably, only N 2 presents evidence of direct laser excitation. This direct excitation produces ionic nitrogen fragments with inner-valence (2s) holes, two unidentified transitions, and one molecular transition, the N 2 + :X 2 Σ g + 2 Σ u + . We discuss these results in the light of a recently proposed model for multiphoton excitation

  1. Characterizing multi-photon quantum interference with practical light sources and threshold single-photon detectors

    Science.gov (United States)

    Navarrete, Álvaro; Wang, Wenyuan; Xu, Feihu; Curty, Marcos

    2018-04-01

    The experimental characterization of multi-photon quantum interference effects in optical networks is essential in many applications of photonic quantum technologies, which include quantum computing and quantum communication as two prominent examples. However, such characterization often requires technologies which are beyond our current experimental capabilities, and today's methods suffer from errors due to the use of imperfect sources and photodetectors. In this paper, we introduce a simple experimental technique to characterize multi-photon quantum interference by means of practical laser sources and threshold single-photon detectors. Our technique is based on well-known methods in quantum cryptography which use decoy settings to tightly estimate the statistics provided by perfect devices. As an illustration of its practicality, we use this technique to obtain a tight estimation of both the generalized Hong‑Ou‑Mandel dip in a beamsplitter with six input photons and the three-photon coincidence probability at the output of a tritter.

  2. Semiclassical analysis of long-wavelength multiphoton processes: The periodically driven harmonic oscillator

    International Nuclear Information System (INIS)

    Fox, Ronald F.; Vela-Arevalo, Luz V.

    2002-01-01

    The problem of multiphoton processes for intense, long-wavelength irradiation of atomic and molecular electrons is presented. The recently developed method of quasiadiabatic time evolution is used to obtain a nonperturbative analysis. When applied to the standard vector potential coupling, an exact auxiliary equation is obtained that is in the electric dipole coupling form. This is achieved through application of the Goeppert-Mayer gauge. While the analysis to this point is general and aimed at microwave irradiation of Rydberg atoms, a Floquet analysis of the auxiliary equation is presented for the special case of the periodically driven harmonic oscillator. Closed form expressions for a complete set of Floquet states are obtained. These are used to demonstrate that for the oscillator case there are no multiphoton resonances

  3. Identification of normal and cancerous human colorectal muscularis propria by multiphoton microscopy in different sections

    Science.gov (United States)

    Zhou, Yi; Chen, Zhifen; Kang, Deyong; li, Lianhuang; Zhuo, Shuangmu; Zhu, Xiaoqin; Guan, Guoxian; Chen, Jianxin

    2016-01-01

    Multiphoton microscopy (MPM) based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) as a potential diagnostic tool is attractive. MPM can effectively provide information about morphological and biochemical changes in biological tissues at the molecular level. In this paper, we attempt to identify normal and cancerous human colorectal muscularis propria by multiphoton microscopy in different sections (both in transverse and longitudinal sections). The results show that MPM can display different microstructure changes in the transverse and longitudinal sections of colorectal muscularis propria. MPM also can quantitatively describe the alteration of collagen content between normal and cancerous muscle layers. These are important pathological findings that MPM images can bring more detailed complementary information about tissue architecture and cell morphology through observing the transverse and longitudinal sections of colorectal muscularis propria. This work demonstrates that MPM can be better for identifying the microstructural characteristics of normal and cancerous human colorectal muscularis propria in different sections.

  4. Multi-photon transitions and Rabi resonance in continuous wave EPR.

    Science.gov (United States)

    Saiko, Alexander P; Fedaruk, Ryhor; Markevich, Siarhei A

    2015-10-01

    The study of microwave-radiofrequency multi-photon transitions in continuous wave (CW) EPR spectroscopy is extended to a Rabi resonance condition, when the radio frequency of the magnetic-field modulation matches the Rabi frequency of a spin system in the microwave field. Using the non-secular perturbation theory based on the Bogoliubov averaging method, the analytical description of the response of the spin system is derived for all modulation frequency harmonics. When the modulation frequency exceeds the EPR linewidth, multi-photon transitions result in sidebands in absorption EPR spectra measured with phase-sensitive detection at any harmonic. The saturation of different-order multi-photon transitions is shown to be significantly different and to be sensitive to the Rabi resonance. The noticeable frequency shifts of sidebands are found to be the signatures of this resonance. The inversion of two-photon lines in some spectral intervals of the out-of-phase first-harmonic signal is predicted under passage through the Rabi resonance. The inversion indicates the transition from absorption to stimulated emission or vice versa, depending on the sideband. The manifestation of the primary and secondary Rabi resonance is also demonstrated in the time evolution of steady-state EPR signals formed by all harmonics of the modulation frequency. Our results provide a theoretical framework for future developments in multi-photon CW EPR spectroscopy, which can be useful for samples with long spin relaxation times and extremely narrow EPR lines. Copyright © 2015 Elsevier Inc. All rights reserved.

  5. Effects of uniform dc electric fields on multiphoton ionization of cesium atoms

    International Nuclear Information System (INIS)

    Klots, C.E.; Compton, R.N.

    1985-01-01

    Multiphoton ionization of cesium atoms shows pronounced two-photon resonances at the nd states and, to a much smaller extent, at the ns states. A dc electric field augments the ns resonances and, for a complementary reason, induces resonances at the np and nf levels. A scaling law for field-induced signals, as a function of principal quantum number, is reported. Field ionization of high Rydberg states is also conveniently studied and quantified with our technique

  6. Multiphoton ionization and fragmentation study of acetone using 308 nm laser radiation

    Science.gov (United States)

    Liu Houxiang, Li Shutao, Han Jingcheng, Zhu Rong, Guan Yifu, Wu Cunkai

    1988-10-01

    Multiphoton ionization and fragmentation (MPI-F) of acetone molecules using 308 nm laser radiation was studied by using a molecular beam and quadrupole mass spectrometer. The ion peaks of acetone molecule appear at m/e=15 and 43, corresponding to the two fragments CH3+ and CH3CO+. It is considered that these two ions are, respectively, formed by direct (2+1) and 2-photon ionization of methyl and acetyl radicals, generated by photodissociation of acetone molecule.

  7. Examination of excited state populations in sputtering using Multiphoton Resonance Ionization

    International Nuclear Information System (INIS)

    Kimock, F.M.; Baxter, J.P.; Pappas, D.L.; Kobrin, P.H.; Winograd, N.

    1984-01-01

    Multiphoton Resonance Ionization has been employed to study the populations of excited state atoms ejected from ion bombarded metal surfaces. Preliminary investigations have focused on three model systems: aluminum, indium and cobalt. In this paper the authors examine the effect of primary ion energy (2 to 12 keV Ar + ) on excited state yields for these three systems. The influence of the sample matrix on excited state populations of sputtered atoms is also discussed

  8. Examination of excited state populations in sputtering using multiphoton resonance ionization

    International Nuclear Information System (INIS)

    Kimock, F.M.; Baxter, J.P.; Pappas, D.L.; Kobrin, P.H.; Winograd, N.

    1984-01-01

    Multiphoton Resonance Ionization has been employed to study the populations of excited state atoms ejected from ion bombarded metal surfaces. Preliminary investigations have focused on three model systems: aluminum, indium and cobalt. In this paper we examine the effect of primary ion energy (2 to 12 keV Ar + ) on excited state yields for these three systems. The influence of the sample matrix on excited state populations of sputtered atoms is also discussed. 8 refs., 4 figs

  9. Star-shaped ladder-type ter(p-phenylene)s for efficient multiphoton absorption.

    Science.gov (United States)

    Guo, Lei; Li, King Fai; Wong, Man Shing; Cheah, Kok Wai

    2013-05-04

    Star-shaped ladder-type ter(p-phenylene)s exhibit remarkably efficient multiphoton absorption properties with 2PA cross-section up to 2579 GM at 700 nm and 3PA cross-section up to 3.35 × 10(-76) cm(6) s(2) in the femtosecond regime for a blue-emissive molecule despite having such a short π-conjugated framework.

  10. Yield and enrichment studies of C-13 isotope by multi-photon ...

    Indian Academy of Sciences (India)

    Abstract. Multi-photon dissociation of Freon-22 (CF2HCl) at low temperatures has been carried out to separate the C-13 isotope using a TEA CO2 laser. Yield and enrichment of C-13 isotope in the product C2F4 are studied at 9P(22) laser line as a function of temperature (–50°C to 30°C). It is observed that at a given fluence ...

  11. Roles of Tunneling, Multiphoton Ionization, and Cascade Ionization for Femtosecond Optical Breakdown in Aqueous Media

    Science.gov (United States)

    2009-09-01

    This implies that a kinetic energy of Ecrit = 1.5 ~ is required for impact ionization [Kai00, Ret04]. The excess energy of 0.5  ~ that remains... Ecrit because the impact ioni- zation rate increases with kinetic energy [Arn92, Kai00, Kel60, Ret04]. To consider both factors, we assume that the...but then neglect it in their theoretical treatment of breakdown [Spa81]. The first comprehensive rate equation model including multiphoton and

  12. Multiphoton microscopy in every lab: the promise of ultrafast semiconductor disk lasers

    Science.gov (United States)

    Emaury, Florian; Voigt, Fabian F.; Bethge, Philipp; Waldburger, Dominik; Link, Sandro M.; Carta, Stefano; van der Bourg, Alexander; Helmchen, Fritjof; Keller, Ursula

    2017-07-01

    We use an ultrafast diode-pumped semiconductor disk laser (SDL) to demonstrate several applications in multiphoton microscopy. The ultrafast SDL is based on an optically pumped Vertical External Cavity Surface Emitting Laser (VECSEL) passively mode-locked with a semiconductor saturable absorber mirror (SESAM) and generates 170-fs pulses at a center wavelength of 1027 nm with a repetition rate of 1.63 GHz. We demonstrate the suitability of this laser for structural and functional multiphoton in vivo imaging in both Drosophila larvae and mice for a variety of fluorophores (including mKate2, tdTomato, Texas Red, OGB-1, and R-CaMP1.07) and for endogenous second-harmonic generation in muscle cell sarcomeres. We can demonstrate equivalent signal levels compared to a standard 80-MHz Ti:Sapphire laser when we increase the average power by a factor of 4.5 as predicted by theory. In addition, we compare the bleaching properties of both laser systems in fixed Drosophila larvae and find similar bleaching kinetics despite the large difference in pulse repetition rates. Our results highlight the great potential of ultrafast diode-pumped SDLs for creating a cost-efficient and compact alternative light source compared to standard Ti:Sapphire lasers for multiphoton imaging.

  13. Multiphoton effects in laser-assisted ionization of a helium atom by electron impact

    Energy Technology Data Exchange (ETDEWEB)

    Ghosh Deb, S.; Sinha, C. [Department of Theoretical Physics, Indian association for the Cultivation of Science, Jadavpur, Kokata (India)

    2010-11-15

    The dynamics of the electron impact multiphoton ionization of a He atom in the presence of an intense laser field (n{gamma}{sub e}, 2e) is studied theoretically for laser polarization (||{sup l}) and perpendicular to the incident momentum. The triple differential (TDCS) as well as the double differential (DDCS) cross sections are studied for the coplanar asymmetric geometry. The results are compared with the only available kinematically complete experiment at high incident energy (1000 eV). Significant laser modification (enhancement) is noted due to multiphoton effects in the present binary and recoil peak intensities of the TDCS for both the geometries, in qualitative agreement with the experiment. In the single photon case, the net effect of the laser field is to suppress the field free (FF) TDCS as well as the DDCS in the zeroth order approximation of the ejected electron wave function (CV), while in the first order (MCV), the cross sections are found to be enhanced. The CV multiphoton cross sections obey the famous Kroll Watson (KW) sum rule while the latter does not hold good in the corresponding MCV approximation. (authors)

  14. Focal switching of photochromic fluorescent proteins enables multiphoton microscopy with superior image contrast.

    Science.gov (United States)

    Kao, Ya-Ting; Zhu, Xinxin; Xu, Fang; Min, Wei

    2012-08-01

    Probing biological structures and functions deep inside live organisms with light is highly desirable. Among the current optical imaging modalities, multiphoton fluorescence microscopy exhibits the best contrast for imaging scattering samples by employing a spatially confined nonlinear excitation. However, as the incident laser power drops exponentially with imaging depth into the sample due to the scattering loss, the out-of-focus background eventually overwhelms the in-focus signal, which defines a fundamental imaging-depth limit. Herein we significantly improve the image contrast for deep scattering samples by harnessing reversibly switchable fluorescent proteins (RSFPs) which can be cycled between bright and dark states upon light illumination. Two distinct techniques, multiphoton deactivation and imaging (MPDI) and multiphoton activation and imaging (MPAI), are demonstrated on tissue phantoms labeled with Dronpa protein. Such a focal switch approach can generate pseudo background-free images. Conceptually different from wave-based approaches that try to reduce light scattering in turbid samples, our work represents a molecule-based strategy that focused on imaging probes.

  15. In vivo multiphoton and fluorescence lifetime imaging microscopy of the healthy and cholestatic liver

    Science.gov (United States)

    Kuznetsova, Daria S.; Dudenkova, Varvara V.; Rodimova, Svetlana A.; Bobrov, Nikolai V.; Zagainov, Vladimir E.; Zagaynova, Elena V.

    2018-02-01

    A cholestatic liver disease presents one of the most common liver diseases and can potentially progress to cirrhosis or even cholangiocarcinoma. Conventional techniques are insufficient to precisely describe the complex internal structure, heterogeneous cell populations and the dynamics of biological processes of the liver. Currently, the methods of multiphoton and fluorescence lifetime imaging microscopy are actively introducing to biomedical research. Those methods are extremely informative and non-destructive that allows studying of a large number of processes occurring inside cells and tissues, analyzing molecular cellular composition, as well as evaluating the state of connective tissue fibers due to their ability to generate a second optical harmonic. Multiphoton and FLIM microscopy do not need additional staining of samples or the incorporation of any markers to study metabolism, lipid composition, microstructure analysis, evaluation of fibrous structures. These parameters have pronounced changes in hepatocytes of liver with common pathological diseases. Thereby in this study we investigated metabolic changes in the healthy and cholestatic liver based on the fluorescence of the metabolic co-factors NAD(P)H and FAD by multiphoton microscopy combined with FLIM. To estimate the contribution of energy metabolism and lipogenesis in the observed changes of the metabolic profile, a separate analysis of NADH and NADPH was presented. The data can be used to develop new criteria for the identification of hepatic pathology at the level of hepatocyte changes directed to personalized medicine in the future.

  16. Polarization control of intermediate state absorption in resonance-mediated multi-photon absorption process

    International Nuclear Information System (INIS)

    Xu, Shuwu; Yao, Yunhua; Jia, Tianqing; Ding, Jingxin; Zhang, Shian; Sun, Zhenrong; Huang, Yunxia

    2015-01-01

    We theoretically and experimentally demonstrate the control of the intermediate state absorption in an (n + m) resonance-mediated multi-photon absorption process by the polarization-modulated femtosecond laser pulse. An analytical solution of the intermediate state absorption in a resonance-mediated multi-photon absorption process is obtained based on the time-dependent perturbation theory. Our theoretical results show that the control efficiency of the intermediate state absorption by the polarization modulation is independent of the laser intensity when the transition from the intermediate state to the final state is coupled by the single-photon absorption, but will be affected by the laser intensity when this transition is coupled by the non-resonant multi-photon absorption. These theoretical results are experimentally confirmed via a two-photon fluorescence control in (2 + 1) resonance-mediated three-photon absorption of Coumarin 480 dye and a single-photon fluorescence control in (1 + 2) resonance-mediated three-photon absorption of IR 125 dye. (paper)

  17. Clinical optical coherence tomography combined with multiphoton tomography for evaluation of several skin disorders

    Science.gov (United States)

    König, Karsten; Speicher, Marco; Bückle, Rainer; Reckfort, Julia; McKenzie, Gordon; Welzel, Julia; Koehler, Martin J.; Elsner, Peter; Kaatz, Martin

    2010-02-01

    The first clinical trial of optical coherence tomography (OCT) combined with multiphoton tomography (MPT) and dermoscopy is reported. State-of-the-art (i) OCT systems for dermatology (e.g. multibeam swept source OCT), (ii) the femtosecond laser multiphoton tomograph DermaInspectTM, and (iii) digital dermoscopes were applied to 47 patients with a diversity of skin diseases and disorders such as skin cancer, psoriasis, hemangioma, connective tissue diseases, pigmented lesions, and autoimmune bullous skin diseases. Dermoscopy, also called 'epiluminescent microscopy', provides two-dimensional color images of the skin surface. OCT imaging is based on the detection of optical reflections within the tissue measured interferometrically whereas nonlinear excitation of endogenous fluorophores and the second harmonic generation are the bases of MPT images. OCT cross sectional "wide field" image provides a typical field of view of 5 x 2 mm2 and offers fast information on the depth and the volume of the investigated lesion. In comparison, multiphoton tomography presents 0.36 x 0.36 mm2 horizontal or diagonal sections of the region of interest within seconds with submicron resolution and down to a tissue depth of 200 μm. The combination of OCT and MPT provides a synergistic optical imaging modality for early detection of skin cancer and other skin diseases.

  18. Visualizing the Acute Effects of Vascular-Targeted Therapy In Vivo Using Intravital Microscopy and Magnetic Resonance Imaging: Correlation with Endothelial Apoptosis, Cytokine Induction, and Treatment Outcome

    Directory of Open Access Journals (Sweden)

    Mukund Seshadri

    2007-02-01

    Full Text Available The acute effects of the vascular-disrupting agent 5,6-dimethylxanthenone-4-acetic acid (DMXAA were investigated in vivo using intravital microscopy (IVM and magnetic resonance imaging (MRI. Changes in vascular permeability and blood flow of syngeneic CT-26 murine colon adenocarcinomas were assessed at 4 and 24 hours after DMXAA treatment (30 mg/kg, i.p. and correlated with induction of tumor necrosis factor-α (TNF-α, endothelial damage [CD31/terminal deoxynucleotidyl transferase (TdT], and treatment outcome. Intravital imaging revealed a marked increase in vascular permeability 4 hours after treatment, consistent with increases in intratumoral mRNA and protein levels of TNF-α. Parallel contrast-enhanced MRI studies showed a ~ 4-fold increase in longitudinal relaxation rates (ΔR1, indicative of increased contrast agent accumulation within the tumor. Dualimmunostained tumor sections (CD31/TdT revealed evidence of endothelial apoptosis at this time point. Twenty-four hours after treatment, extensive hemorrhage and complete disruption of vascular architecture were observed with IVM, along with a significant reduction in ΔR1 and virtual absence of CD31 immunostaining. DMXAA-induced tumor vascular damage resulted in significant long-term (60-day cures compared to untreated controls. Multimodality imaging approaches are useful in visualizing the effects of antivascular therapy in vivo. Such approaches allow cross validation and correlation of findings with underlying molecular changes contributing to treatment outcome.

  19. Where are we? The anatomy of the murine cortical meninges revisited for intravital imaging, immunology, and clearance of waste from the brain.

    Science.gov (United States)

    Coles, Jonathan A; Myburgh, Elmarie; Brewer, James M; McMenamin, Paul G

    2017-09-01

    Rapid progress is being made in understanding the roles of the cerebral meninges in the maintenance of normal brain function, in immune surveillance, and as a site of disease. Most basic research on the meninges and the neural brain is now done on mice, major attractions being the availability of reporter mice with fluorescent cells, and of a huge range of antibodies useful for immunocytochemistry and the characterization of isolated cells. In addition, two-photon microscopy through the unperforated calvaria allows intravital imaging of the undisturbed meninges with sub-micron resolution. The anatomy of the dorsal meninges of the mouse (and, indeed, of all mammals) differs considerably from that shown in many published diagrams: over cortical convexities, the outer layer, the dura, is usually thicker than the inner layer, the leptomeninx, and both layers are richly vascularized and innervated, and communicate with the lymphatic system. A membrane barrier separates them and, in disease, inflammation can be localized to one layer or the other, so experimentalists must be able to identify the compartment they are studying. Here, we present current knowledge of the functional anatomy of the meninges, particularly as it appears in intravital imaging, and review their role as a gateway between the brain, blood, and lymphatics, drawing on information that is scattered among works on different pathologies. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Application of Negative Curvature Hollow-Core Fiber in an Optical Fiber Sensor Setup for Multiphoton Spectroscopy.

    Science.gov (United States)

    Popenda, Maciej Andrzej; Stawska, Hanna Izabela; Mazur, Leszek Mateusz; Jakubowski, Konrad; Kosolapov, Alexey; Kolyadin, Anton; Bereś-Pawlik, Elżbieta

    2017-10-06

    In this paper, an application of negative curvature hollow core fiber (NCHCF) in an all-fiber, multiphoton fluorescence sensor setup is presented. The dispersion parameter (D) of this fiber does not exceed the value of 5 ps/nm × km across the optical spectrum of (680-750) nm, making it well suited for the purpose of multiphoton excitation of biological fluorophores. Employing 1.5 m of this fiber in a simple, all-fiber sensor setup allows us to perform multiphoton experiments without any dispersion compensation methods. Multiphoton excitation of nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) with this fiber shows a 6- and 9-fold increase, respectively, in the total fluorescence signal collected when compared with the commercial solution in the form of a hollow-core photonic band gap fiber (HCPBF). To the author's best knowledge, this is the first time an NCHCF was used in an optical-fiber sensor setup for multiphoton fluorescence experiments.

  1. In vivo multiphoton imaging of human skin: assessment of topical corticosteroid-induced epidermis atrophy and depigmentation

    Science.gov (United States)

    Ait El Madani, Hassan; Tancrède-Bohin, Emmanuelle; Bensussan, Armand; Colonna, Anne; Dupuy, Alain; Bagot, Martine; Pena, Ana-Maria

    2012-02-01

    Multiphoton microscopy has emerged in the past decade as a promising tool for noninvasive skin imaging. Our aim was to evaluate the potential of multiphoton microscopy to detect topical corticosteroids side effects within the epidermis and to provide new insights into their dynamics. Healthy volunteers were topically treated with clobetasol propionate on a small region of their forearms under overnight occlusion for three weeks. The treated region of each patient was investigated at D0, D7, D15, D22 (end of the treatment), and D60. Our study shows that multiphoton microscopy allows for the detection of corticoid-induced epidermis modifications: thinning of stratum corneum compactum and epidermis, decrease of keratinocytes size, and changes in their morphology from D7 to D22. We also show that multiphoton microscopy enables in vivo three-dimensional (3-D) quantitative assessment of melanin content. We observe that melanin density decreases during treatment and almost completely disappears at D22. Moreover, these alterations are reversible as they are no longer present at D60. Our study demonstrates that multiphoton microscopy is a convenient and powerful tool for noninvasive 3-D dynamical studies of skin integrity and pigmentation.

  2. Design and synthesis of hyperstructured molecules based on cyclophosphazene core for multiphoton absorption

    International Nuclear Information System (INIS)

    Naik, K. Praveen Kumar; Sreeramulu, V.; Ramya, E.; Muralidharan, K.; Rao, D. Narayana

    2016-01-01

    Cyclophosphazene based hyperstructured molecules were synthesized through simple nucleophilic substitution reactions. All these molecules were characterized by multinuclear NMR, MALDI and HRMS spectral data. Third order nonlinear optical properties of the hyperstructured molecules were measured using Z-scan technique with 532 nm, picosecond (ps) laser and 800 nm, femtosecond (fs) laser pulses. The molecules showed reverse saturable absorption on excitation at both 532 nm and 800 nm, which could be attributed to the two-photon absorption (2 PA) and three-photon absorption (3 PA), respectively. The 2 PA and 3 PA cross section values exhibited by the molecules based on cyclophosphazene are as high as 527 GM and 1.86 × 10"−"7"6 cm"−"6 s"2 photon"−"1, respectively. The 2PA, 3PA coefficients and optical limiting properties make them suitable candidates for nonlinear optical devices in the visible and near IR range. - Graphical abstract: The hyperstructured molecules based on cyclophosphazene core were synthesized and used for multiphoton absorption. Open aperture Z-scan curves of hyper structured molecules at the excitation of (a) picosecond laser and (b) femtosecond laser representing multiphoton absorption properties are reported. - Highlights: • Two hyperstructured molecules based on cyclophosphazene core are designed for multiphoton absorption. • NLO properties are measured using Z-scan technique at 532 nm and 800 nm wavelengths. • The molecules were tested for the optical limiting applications at 532 nm and 800 nm laser pulses.

  3. Effects of multi-photon interferences from internally generated fields in strongly resonant systems

    International Nuclear Information System (INIS)

    Deng, Lu; Payne, Marvin G.; Garrett, William R.

    2006-01-01

    In studies of various nonlinear optical phenomena, strong resonant features in the atomic or molecular response to multi-photon driven processes have been used to greatly enhance the visibility of otherwise weak higher-order processes. However, there are well defined circumstances where a multi-photon-resonant response of a target system leads to the generation of one or more new electromagnetic fields that can drastically change the overall system response from what would be expected from the imposed laser fields alone. New effects can occur and dominate some aspects of the nonlinear optical response because of the constructive or destructive interference between transition amplitudes along multiple excitation pathways between a given set of optically coupled states, where one of the pathways involve internally generated field(s). Under destructive interference some resonant enhancements can become completely canceled (suppressed). This review focuses on the class of optical interference effects associated with internally generated fields, that have been found to be capable of influencing a very significant number of basic physical phenomena in gas or vapor phase systems. It provides a historical overview of experimental and theoretical developments and a modern understanding of the underlying physics and its various manifestations that include: suppression of multi-photon excitation processes, suppression of stimulated emissions (Raman, hyper-Raman, and optically pumped stimulated emissions), saturation of parametric wave-mixing, pressure and beam-geometry dependent shifting of multi-photon-resonant absorption lines, and the suppression of Autler-Townes splitting and ac-stark shifts. Additionally, optical interference effects in some modern contexts, such as achieving multi-photon induced transparency, establishing single-photon self-interference based induced transparency, and generating entangled single photon states, are reviewed

  4. Design and synthesis of hyperstructured molecules based on cyclophosphazene core for multiphoton absorption

    Energy Technology Data Exchange (ETDEWEB)

    Naik, K. Praveen Kumar [School of Chemistry, University of Hyderabad, Hyderabad 500046 India (India); Sreeramulu, V. [School of Physics, University of Hyderabad, Hyderabad 500046 India (India); CNR-IFN CSMFO Laboratory, Via alla Cascata, 56/C Povo, Trento (Italy); Ramya, E. [School of Physics, University of Hyderabad, Hyderabad 500046 India (India); Muralidharan, K., E-mail: murali@uohyd.ac.in [School of Chemistry, University of Hyderabad, Hyderabad 500046 India (India); Rao, D. Narayana [School of Physics, University of Hyderabad, Hyderabad 500046 India (India)

    2016-09-01

    Cyclophosphazene based hyperstructured molecules were synthesized through simple nucleophilic substitution reactions. All these molecules were characterized by multinuclear NMR, MALDI and HRMS spectral data. Third order nonlinear optical properties of the hyperstructured molecules were measured using Z-scan technique with 532 nm, picosecond (ps) laser and 800 nm, femtosecond (fs) laser pulses. The molecules showed reverse saturable absorption on excitation at both 532 nm and 800 nm, which could be attributed to the two-photon absorption (2 PA) and three-photon absorption (3 PA), respectively. The 2 PA and 3 PA cross section values exhibited by the molecules based on cyclophosphazene are as high as 527 GM and 1.86 × 10{sup −76} cm{sup −6} s{sup 2} photon{sup −1}, respectively. The 2PA, 3PA coefficients and optical limiting properties make them suitable candidates for nonlinear optical devices in the visible and near IR range. - Graphical abstract: The hyperstructured molecules based on cyclophosphazene core were synthesized and used for multiphoton absorption. Open aperture Z-scan curves of hyper structured molecules at the excitation of (a) picosecond laser and (b) femtosecond laser representing multiphoton absorption properties are reported. - Highlights: • Two hyperstructured molecules based on cyclophosphazene core are designed for multiphoton absorption. • NLO properties are measured using Z-scan technique at 532 nm and 800 nm wavelengths. • The molecules were tested for the optical limiting applications at 532 nm and 800 nm laser pulses.

  5. Characterizing germania concentration and structure in fiber soot using multiphoton microscopy and spectroscopy technology

    Science.gov (United States)

    Chen, Minghan; Li, Ming-Jun; Liu, Anping

    2015-02-01

    Germania doping is commonly used in the core of optical fiber due to its advantages compared to other materials such as superior transparency in near-infrared telecommunication wavelength region. During fiber preform manufacturing using the outside vapor deposition (OVD) process, Ge is doped into a silica soot preform by chemical vapor deposition. Since the Ge doping concentration profile is directly correlated with the fiber refractive index profile, its characterization is critical for the fiber industry. Electron probe micro-analyzer (EPMA) is a conventional analysis method for characterizing the Ge concentration profile. However, it requires extensive sample preparation and lengthy measurement. In this paper, a multiphoton microscopy technique is utilized to measure the Ge doping profile based on the multiphoton fluorescence intensity of the soot layers. Two samples, one with ramped and another with stepped Ge doping profiles were prepared for measurements. Measured results show that the technique is capable of distinguishing ramped and stepped Ge doping profiles with good accuracy. In the ramped soot sample, a sharp increment of doping level was observed in about 2 mm range from soot edge followed by a relative slow gradient doping accretion. As for the stepped doping sample, step sizes ranging from around 1 mm (at soot edge) to 3 mm (at soot center) were observed. All the measured profiles are in close agreement with that of the EPMA measurements. In addition, both multiphoton fluorescence (around 420 nm) and sharp second harmonic generations (at 532 nm) were observed, which indicates the co-existence of crystal and amorphous GeO2.

  6. High Resolution Multiphoton Ionization/Dissociation of Molecular Beam of Acetone from 582.60 to 585.80 nm

    Science.gov (United States)

    Mejia-Ospino, Enrique; Garcia, Gladis; Guerrero, Alfonso; Alvarez, Ignacio; Cisneros, Carmen

    2003-05-01

    Multiphoton ionization and dissociation of a jet supersonic of acetone at wavelength from 582.60 to 585.80 nm is studied using a Nd:YAG-OPO (optical parametric oscillator) system coupled to time-of-flight mass spectrometer. We present high-resolution (1.5 cm-1) three-photon resonance multiphoton spectra of the acetone 3s CH3CO+), (CH3+) and (COH+), being CH3COCH3+ ---> CH3CO+ + CH3 the more likely channel. The molecular and acetyl ions are observed practically in overall wavelength range.

  7. Design, Fabrication and Computational Characterization of a 3D Micro-Valve Built by Multi-Photon Polymerization

    Directory of Open Access Journals (Sweden)

    Stratos Galanopoulos

    2014-08-01

    Full Text Available We report on the design, modeling and fabrication by multi-photon polymerization of a complex medical fluidic device. The physical dimensions of the built micro-valve prototype are compared to those of its computer-designed model. Important fabrication issues such as achieving high dimensional resolution and ability to control distortion due to shrinkage are presented and discussed. The operational performance of both multi-photon and CAD-created models under steady blood flow conditions was evaluated and compared through computational fluid dynamics analysis.

  8. Influence of evanescent waves on the voxel profile in multipulse multiphoton polymerization nanofabrication

    International Nuclear Information System (INIS)

    Li Wei; Cao Tianxiang; Zhai Zhaohui; Yu Xuanyi; Zhang Xinzheng; Xu Jingjun

    2013-01-01

    The relationship between the profile of the structures obtained by multiphoton polymerization and the optical parameters of nanofabrication systems has been studied theoretically for a multipulse scheme. We find that the profile of sub-wavelength structures is greatly affected by the evanescent waves affect. Not only is the photocured polymer voxel affected by the beam profile, but the beam propagation behavior is influenced by the photocured polymer voxel. This gives us a new view of matter–light interactions in multipulse polymerization process, which is useful to the accurate control of the nanofabrication profile and the selection of new nanofabrication materials. (paper)

  9. Waveguide-integrated single- and multi-photon detection at telecom wavelengths using superconducting nanowires

    International Nuclear Information System (INIS)

    Ferrari, Simone; Kahl, Oliver; Kovalyuk, Vadim; Goltsman, Gregory N.; Korneev, Alexander; Pernice, Wolfram H. P.

    2015-01-01

    We investigate single- and multi-photon detection regimes of superconducting nanowire detectors embedded in silicon nitride nanophotonic circuits. At near-infrared wavelengths, simultaneous detection of up to three photons is observed for 120 nm wide nanowires biased far from the critical current, while narrow nanowires below 100 nm provide efficient single photon detection. A theoretical model is proposed to determine the different detection regimes and to calculate the corresponding internal quantum efficiency. The predicted saturation of the internal quantum efficiency in the single photon regime agrees well with plateau behavior observed at high bias currents

  10. Waveguide-integrated single- and multi-photon detection at telecom wavelengths using superconducting nanowires

    Energy Technology Data Exchange (ETDEWEB)

    Ferrari, Simone; Kahl, Oliver [Institute of Nanotechnology, Karlsruhe Institute of Technology, Karlsruhe 76132 (Germany); Kovalyuk, Vadim [Institute of Nanotechnology, Karlsruhe Institute of Technology, Karlsruhe 76132 (Germany); Department of Physics, Moscow State Pedagogical University, Moscow 119992 (Russian Federation); Goltsman, Gregory N. [Department of Physics, Moscow State Pedagogical University, Moscow 119992 (Russian Federation); National Research University Higher School of Economics, 20 Myasnitskaya Ulitsa, Moscow 101000 (Russian Federation); Korneev, Alexander [Department of Physics, Moscow State Pedagogical University, Moscow 119992 (Russian Federation); Moscow Institute of Physics and Technology (State University), Moscow 141700 (Russian Federation); Pernice, Wolfram H. P., E-mail: wolfram.pernice@kit.edu [Institute of Nanotechnology, Karlsruhe Institute of Technology, Karlsruhe 76132 (Germany); Department of Physics, University of Münster, 48149 Münster (Germany)

    2015-04-13

    We investigate single- and multi-photon detection regimes of superconducting nanowire detectors embedded in silicon nitride nanophotonic circuits. At near-infrared wavelengths, simultaneous detection of up to three photons is observed for 120 nm wide nanowires biased far from the critical current, while narrow nanowires below 100 nm provide efficient single photon detection. A theoretical model is proposed to determine the different detection regimes and to calculate the corresponding internal quantum efficiency. The predicted saturation of the internal quantum efficiency in the single photon regime agrees well with plateau behavior observed at high bias currents.

  11. Athermal electron distribution probed by femtosecond multiphoton photoemission from image potential states

    International Nuclear Information System (INIS)

    Ferrini, Gabriele; Giannetti, Claudio; Pagliara, Stefania; Banfi, Francesco; Galimberti, Gianluca; Parmigiani, Fulvio

    2005-01-01

    Image potential states are populated through indirect, scattering-mediated multiphoton absorption induced by femtosecond laser pulses and revealed by single-photon photoemission. The measured effective mass is significantly different from that obtained with direct, resonant population. These features reveal a strong coupling of the electrons residing in the image potential state, outside the solid, with the underlying hot electron population created by the laser pulse. The coupling is mediated by a many-body scattering interaction between the image potential state electrons and bulk electrons in highly excited states

  12. Experimental quantum teleportation and multiphoton entanglement via interfering narrowband photon sources

    International Nuclear Information System (INIS)

    Yang Jian; Zhang Han; Peng Chengzhi; Chen Zengbing; Bao Xiaohui; Chen Shuai; Pan Jianwei

    2009-01-01

    In this paper, we report a realization of synchronization-free quantum teleportation and narrowband three-photon entanglement through interfering narrowband photon sources. Since both the single-photon and the entangled photon pair utilized are completely autonomous, it removes the requirement of high-demanding synchronization techniques in long-distance quantum communication with pulsed spontaneous parametric down-conversion sources. The frequency linewidth of the three-photon entanglement realized is on the order of several MHz, which matches the requirement of atomic ensemble based quantum memories. Such a narrowband multiphoton source will have applications in some advanced quantum communication protocols and linear optical quantum computation.

  13. Atomic-structure effects in strong-field multiphoton detachment and ionization

    International Nuclear Information System (INIS)

    AAberg, T.; Mu, X.; Ruscheinski, J.; Crasemann, B.

    1994-01-01

    Above-threshold photoelectron detachment and ionization spectra are investigated theoretically in the tunneling and over-barrier regime as a function of wavelength (≥ 1.064 μm) and polarization of the electromagnetic field. It is found that the zeros in the initial-state wave function can drastically affect the shape of the high-energy photoelectron distribution. The phenomenon is not predicted by tunneling and related models and hence can test their validity and reveal whether Keldysh-type theories are in general applicable to strong-field multiphoton dynamics. (orig.)

  14. A multiphoton objective design with incorporated beam splitter for enhanced fluorescence collection.

    Science.gov (United States)

    McMullen, Jesse D; Zipfel, Warren R

    2010-03-15

    We present a de novo design of an objective for use in multi-photon (MPM) and second harmonic generation (SHG) microscopy. This objective was designed to have a large field of view (FOV), while maintaining a moderate numerical aperture (NA) and relative straight forward construction. A dichroic beam splitter was incorporated within the objective itself allowing for an increase in the front aperture of the objective and corresponding enhancement of the solid angle of collected emission by an order of magnitude over existing designs.

  15. On the separation of enantiomers of 1,1,1,2-tetrafluoroiodoethane by IR multiphoton excitation

    International Nuclear Information System (INIS)

    Pochert, J.; Quack, M.; Seyfang, G.

    2002-01-01

    The first attempt to separate enantiomers of chiral molecules using IR-multiphoton excitation with circularly polarized light is reported. 1,1,1,2-Tetrafuoroiodoethane CF 3 CHFI has been chosen as its IR-spectroscopy and its IR-photochemistry is well characterized by our previous work. A theoretical model based on a master equation is presented to predict the enrichment factor. The experimental results show that the experimental sensitivity must be improved to reach the limit of the theoretical prediction. (author)

  16. Microwave multiphoton excitation of helium Rydberg atoms: The analogy with atomic collisions

    International Nuclear Information System (INIS)

    van de Water, W.; van Leeuwen, K.A.H.; Yoakum, S.; Galvez, E.J.; Moorman, L.; Bergeman, T.; Sauer, B.E.; Koch, P.M.

    1989-01-01

    We study multiphoton transitions in helium Rydberg atoms subjected to a microwave electric field of fixed frequency but varying intensity. For each principal quantum number in the range n=25--32, the n 3 S to n 3 (L>2), n=25--32, transition probability exhibits very sharp structures as a function of the field amplitude. Their positions could be reproduced precisely using a Floquet Hamiltonian for the interaction between atom and field. Their shapes are determined by the transients of field turn-on and turn-off in a way that makes a close analogy with the theory of slow atomic collisions

  17. Ionisation of hydrogen-like atoms by a multiphoton absorption process

    International Nuclear Information System (INIS)

    Gontier, Y.; Trahin, M.

    1967-01-01

    The general expression for the amplitude of the probability of ionisation by a multiphoton absorption process is derived. Its non-relativistic limit is taken and the bipolar approximation is used for calculating the ionisation cross-section of hydrogen-like atoms. This latter involves the summation over intermediate virtual states by means of: a) a recursion relationship concerning angular functions, b) a particular technique which when applied to radial functions makes it possible to solve a system of inhomogeneous first-order differential equations. (authors) [fr

  18. In vivo stepwise multi-photon activation fluorescence imaging of melanin in human skin

    Science.gov (United States)

    Lai, Zhenhua; Gu, Zetong; Abbas, Saleh; Lowe, Jared; Sierra, Heidy; Rajadhyaksha, Milind; DiMarzio, Charles

    2014-03-01

    The stepwise multi-photon activated fluorescence (SMPAF) of melanin is a low cost and reliable method of detecting melanin because the activation and excitation can be a continuous-wave (CW) mode near infrared (NIR) laser. Our previous work has demonstrated the melanin SMPAF images in sepia melanin, mouse hair, and mouse skin. In this study, we show the feasibility of using SMPAF to detect melanin in vivo. in vivo melanin SMPAF images of normal skin and benign nevus are demonstrated. SMPAF images add specificity for melanin detection than MPFM images and CRM images. Melanin SMPAF is a promising technology to enable early detection of melanoma for dermatologists.

  19. Search for anomalous multiphoton production at 100-300 GeV

    International Nuclear Information System (INIS)

    Burke, D.L.; Gustafson, H.R.; Jones, L.W.; Longo, M.J.

    1975-01-01

    A search for anomalous multiphoton production in neutron-CH 2 collisions has been carried out at Fermilab. Both anomalous γ events as might be produced in the annihilation of a magnetic monopole pair, as well as events with smaller opening angles, such as those observed in cosmic ray emulsions by Schein et al. and others were sought. No evidence for either type of event was found. An upper limit approximately 2.7 μb is placed on the production cross section for 'Schein' events or approximately10 -2 that deduced from the cosmic ray data. (Auth.)

  20. Multiphoton ionization of H+2 at critical internuclear separations: non-Hermitian Floquet analysis

    International Nuclear Information System (INIS)

    Likhatov, P V; Telnov, D A

    2009-01-01

    We present ab initio time-dependent non-Hermitian Floquet calculations of multiphoton ionization (MPI) rates of hydrogen molecular ions subject to an intense linearly polarized monochromatic laser field with a wavelength of 800 nm. The orientation of the molecular axis is parallel to the polarization vector of the laser field. The MPI rates are computed for a wide range of internuclear separations R with high resolution in R and reproduce resonance and near-threshold structures. We show that enhancement of ionization at critical internuclear separations is related to resonance series with higher electronic states. The effect of two-centre interference on the MPI signal is discussed.

  1. Resonance-enhanced multiphoton ionization (REMPI) spectroscopy of p-chlorofluorobenzene

    Science.gov (United States)

    Tuttle, William D.; Gardner, Adrian M.; Wright, Timothy G.

    2017-09-01

    The S1 ← S0 (A˜1 B2 ← X˜1 A1) electronic transition of para-chlorofluorobenzene has been investigated using resonance-enhanced multiphoton ionization (REMPI) spectroscopy. Assignment of the vibrational structure has been achieved by comparison with corresponding spectra of related molecules, via quantum chemical calculations, and via shifts in bands between the spectra of the 35Cl and 37Cl isotopologues. In addition, we have also partially reassigned a previously-published spectrum of para-dichlorobenzene.

  2. Periodicity in tumor vasculature targeting kinetics of ligand-functionalized nanoparticles studied by dynamic contrast enhanced magnetic resonance imaging and intravital microscopy

    DEFF Research Database (Denmark)

    Hak, Sjoerd; Cebulla, Jana; Huuse, Else Marie

    2014-01-01

    In the past two decades advances in the development of targeted nanoparticles have facilitated their application as molecular imaging agents and targeted drug delivery vehicles. Nanoparticle-enhanced molecular imaging of the angiogenic tumor vasculature has been of particular interest. Not only...... because angiogenesis plays an important role in various pathologies, but also since endothelial cell surface receptors are directly accessible for relatively large circulating nanoparticles. Typically, nanoparticle targeting towards these receptors is studied by analyzing the contrast distribution...... kinetics. These kinetics will not only depend on nanoparticle characteristics, but also on receptor binding and recycling. In this study, we monitored the in vivo targeting kinetics of αvβ3-integrin specific nanoparticles with intravital microscopy and dynamic contrast enhanced magnetic resonance imaging...

  3. Privacy Act

    Science.gov (United States)

    Learn about the Privacy Act of 1974, the Electronic Government Act of 2002, the Federal Information Security Management Act, and other information about the Environmental Protection Agency maintains its records.

  4. Analyzing Structure and Function of Vascularization in Engineered Bone Tissue by Video-Rate Intravital Microscopy and 3D Image Processing.

    Science.gov (United States)

    Pang, Yonggang; Tsigkou, Olga; Spencer, Joel A; Lin, Charles P; Neville, Craig; Grottkau, Brian

    2015-10-01

    Vascularization is a key challenge in tissue engineering. Three-dimensional structure and microcirculation are two fundamental parameters for evaluating vascularization. Microscopic techniques with cellular level resolution, fast continuous observation, and robust 3D postimage processing are essential for evaluation, but have not been applied previously because of technical difficulties. In this study, we report novel video-rate confocal microscopy and 3D postimage processing techniques to accomplish this goal. In an immune-deficient mouse model, vascularized bone tissue was successfully engineered using human bone marrow mesenchymal stem cells (hMSCs) and human umbilical vein endothelial cells (HUVECs) in a poly (D,L-lactide-co-glycolide) (PLGA) scaffold. Video-rate (30 FPS) intravital confocal microscopy was applied in vitro and in vivo to visualize the vascular structure in the engineered bone and the microcirculation of the blood cells. Postimage processing was applied to perform 3D image reconstruction, by analyzing microvascular networks and calculating blood cell viscosity. The 3D volume reconstructed images show that the hMSCs served as pericytes stabilizing the microvascular network formed by HUVECs. Using orthogonal imaging reconstruction and transparency adjustment, both the vessel structure and blood cells within the vessel lumen were visualized. Network length, network intersections, and intersection densities were successfully computed using our custom-developed software. Viscosity analysis of the blood cells provided functional evaluation of the microcirculation. These results show that by 8 weeks, the blood vessels in peripheral areas function quite similarly to the host vessels. However, the viscosity drops about fourfold where it is only 0.8 mm away from the host. In summary, we developed novel techniques combining intravital microscopy and 3D image processing to analyze the vascularization in engineered bone. These techniques have broad

  5. Hotspot related plasmon assisted multiphoton photocurrents in metal-insulator-metal junctions

    Energy Technology Data Exchange (ETDEWEB)

    Differt, Dominik; Pfeiffer, Walter [Universitaet Bielefeld, Universitaetsstr. 25, 33615 Bielefeld (Germany); Diesing, Detlef [Universitaet Duisburg-Essen, Universitaetsstr. 5, 45117 Essen (Germany)

    2011-07-01

    Scanning photocurrent microscopy of metal-insulator-metal junctions (MIM) is used to investigate the mechanisms of femtosecond multiphoton photocurrent injection at liquid nitrogen temperature. The locally induced multiphoton photocurrent in a Ag-TaO-Ta MIM junction is measured in a scanning microscope cryostat under focused illumination (5{mu}m focus diameter, 800 nm, 30 fs, 80 MHz repetition rate). The intensity dependence reveals a mixture of two-photon and three-photon processes that are responsible for the photocurrent. Its lateral variation shows hotspot-like behaviour with significant magnitude variations on a 100 to 200 nm length scale. Assuming an injection current duration of 40fs the peak injection current density of about 10{sup 4} A cm{sup -2} is estimated - 10{sup 6} times higher than that for 400 nm continuous wave illumination slightly below the damage threshold. The simultaneously measured extinction of the incident radiation reveals a 20 to 30% increased absorption at the hotspots. We attribute the local photocurrent enhancement to the defect-assisted excitation of surface plasmon polaritons at the silver electrode leading to an enhanced local excitation.

  6. Controllable surfaces of path interference in the multiphoton ionization of atoms by a weak trichromatic field

    International Nuclear Information System (INIS)

    Mercouris, Theodoros; Nicolaides, Cleanthes A

    2005-01-01

    Multiphoton detachment rates for the H - 1 S ground state irradiated by a weak trichromatic ac field consisting of the fundamental frequency ω 0.272 eV and its second, third or fourth higher harmonics were computed from first principles. The weak intensities are in the range of 10 7 -10 8 W cm -2 . The calculations incorporated systematically electronic structure and electron correlation effects. They were done by implementing a time-independent, nonperturbative many-electron, many-photon theory (MEMPT) which obtains cycle-averaged complex eigenvalues, whose real part gives the field-induced energy shift, Δ, and the imaginary part is the multiphoton ionization rate, Γ. Through analysis, plausible arguments and computation, we show that when the intensities are weak the dependence of Γ on phase differences is simple. Specifically, Γs are depicted in the form of plane surfaces, with minor ripples due to higher order ionization paths, in terms of trigonometric functions of the phase differences. This dependence is likely to be applicable to other atomic systems as well, and to provide a definition of the weak field regime in the trichromatic case. When the field intensities are such that higher order ionization paths become important, these dependences break down and we reach the strong field regime

  7. A least squares approach to estimating the probability distribution of unobserved data in multiphoton microscopy

    Science.gov (United States)

    Salama, Paul

    2008-02-01

    Multi-photon microscopy has provided biologists with unprecedented opportunities for high resolution imaging deep into tissues. Unfortunately deep tissue multi-photon microscopy images are in general noisy since they are acquired at low photon counts. To aid in the analysis and segmentation of such images it is sometimes necessary to initially enhance the acquired images. One way to enhance an image is to find the maximum a posteriori (MAP) estimate of each pixel comprising an image, which is achieved by finding a constrained least squares estimate of the unknown distribution. In arriving at the distribution it is assumed that the noise is Poisson distributed, the true but unknown pixel values assume a probability mass function over a finite set of non-negative values, and since the observed data also assumes finite values because of low photon counts, the sum of the probabilities of the observed pixel values (obtained from the histogram of the acquired pixel values) is less than one. Experimental results demonstrate that it is possible to closely estimate the unknown probability mass function with these assumptions.

  8. Multiphoton crosslinking for biocompatible 3D printing of type I collagen.

    Science.gov (United States)

    Bell, Alex; Kofron, Matthew; Nistor, Vasile

    2015-09-03

    Multiphoton fabrication is a powerful technique for three-dimensional (3D) printing of structures at the microscale. Many polymers and proteins have been successfully structured and patterned using this method. Type I collagen comprises a large part of the extracellular matrix for most tissue types and is a widely used cellular scaffold material for tissue engineering. Current methods for creating collagen tissue scaffolds do not allow control of local geometry on a cellular scale. This means the environment experienced by cells may be made up of the native material but unrelated to native cellular-scale structure. In this study, we present a novel method to allow multiphoton crosslinking of type I collagen with flavin mononucleotide photosensitizer. The method detailed allows full 3D printing of crosslinked structures made from unmodified type I collagen and uses only demonstrated biocompatible materials. Resolution of 1 μm for both standing lines and high-aspect ratio gaps between structures is demonstrated and complex 3D structures are fabricated. This study demonstrates a means for 3D printing with one of the most widely used tissue scaffold materials. High-resolution, 3D control of the fabrication of collagen scaffolds will facilitate higher fidelity recreation of the native extracellular environment for engineered tissues.

  9. A novel multiphoton microscopy images segmentation method based on superpixel and watershed.

    Science.gov (United States)

    Wu, Weilin; Lin, Jinyong; Wang, Shu; Li, Yan; Liu, Mingyu; Liu, Gaoqiang; Cai, Jianyong; Chen, Guannan; Chen, Rong

    2017-04-01

    Multiphoton microscopy (MPM) imaging technique based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) shows fantastic performance for biological imaging. The automatic segmentation of cellular architectural properties for biomedical diagnosis based on MPM images is still a challenging issue. A novel multiphoton microscopy images segmentation method based on superpixels and watershed (MSW) is presented here to provide good segmentation results for MPM images. The proposed method uses SLIC superpixels instead of pixels to analyze MPM images for the first time. The superpixels segmentation based on a new distance metric combined with spatial, CIE Lab color space and phase congruency features, divides the images into patches which keep the details of the cell boundaries. Then the superpixels are used to reconstruct new images by defining an average value of superpixels as image pixels intensity level. Finally, the marker-controlled watershed is utilized to segment the cell boundaries from the reconstructed images. Experimental results show that cellular boundaries can be extracted from MPM images by MSW with higher accuracy and robustness. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Multiphoton imaging of low grade, high grade intraepithelial neoplasia and intramucosal invasive cancer of esophagus

    Science.gov (United States)

    Xu, Jian; Jiang, Liwei; Kang, Deyong; Wu, Xuejing; Xu, Meifang; Zhuo, Shuangmu; Zhu, Xiaoqin; Lin, Jiangbo; Chen, Jianxin

    2017-04-01

    Esophageal squamous cell carcinoma (ESCC) is devastating because of its aggressive lymphatic spread and clinical course. It is believed to occur through low-grade intraepithelial neoplasia (LGIN), high-grade intraepithelial neoplasia (HGIN), and intramucosal invasive cancer (IMC) before transforming to submucosal cancer. In particular, these early lesions (LGIN, HGIN and IMC), which involve no lymph node nor distant metastasis, can be cured by endoscopic treatment. Therefore, early identification of these lesions is important so as to offer a curative endoscopic resection, thus slowing down the development of ESCC. In this work, spectral information and morphological features of the normal esophageal mucosa are first studied. Then, the morphological changes of LGIN, HGIN and IMC are described. Lastly, quantitative parameters are also extracted by calculating the nuclear-to-cytoplasmic ratio of epithelial cells and the pixel density of collagen in the lamina propria. These results show that multiphoton microscopy (MPM) has the ability to identify normal esophageal mucosa, LGIN, HGIN and IMC. With the development of multiphoton endoscope systems for in vivo imaging, combined with a laser ablation system, MPM has the potential to provide immediate pathologic diagnosis and curative treatment of ESCC before the transformation to submucosal cancer in the future.

  11. Insights on proximity effect and multiphoton induced luminescence from gold nanospheres in far field optical microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Borglin, Johan [Biomedical Photonics Group, Department of Chemistry and Molecular Biology, University of Gothenburg, Kemivägen 10, 412 96 Gothenburg (Sweden); Department of Physics, University of Gothenburg, Kemivägen 10, 412 96 Gothenburg (Sweden); Guldbrand, Stina [Department of Physics, University of Gothenburg, Kemivägen 10, 412 96 Gothenburg (Sweden); Evenbratt, Hanne [Pharmaceutical Technology, Department of Chemistry and Chemical Engineering, Chalmers University of Technology, Kemigården 4, 412 96 Gothenburg (Sweden); Kirejev, Vladimir; Ericson, Marica B., E-mail: marica.ericson@chem.gu.se [Biomedical Photonics Group, Department of Chemistry and Molecular Biology, University of Gothenburg, Kemivägen 10, 412 96 Gothenburg (Sweden); Grönbeck, Henrik [Department of Applied Physics, Chalmers University of Technology, Kemivägen 9, 412 96 Gothenburg (Sweden)

    2015-12-07

    Gold nanoparticles can be visualized in far-field multiphoton laser-scanning microscopy (MPM) based on the phenomena of multiphoton induced luminescence (MIL). This is of interest for biomedical applications, e.g., for cancer diagnostics, as MPM allows for working in the near-infrared (NIR) optical window of tissue. It is well known that the aggregation of particles causes a redshift of the plasmon resonance, but its implications for MIL applying far-field MPM should be further exploited. Here, we explore MIL from 10 nm gold nanospheres that are chemically deposited on glass substrates in controlled coverage gradients using MPM operating in NIR range. The substrates enable studies of MIL as a function of inter-particle distance and clustering. It was shown that MIL was only detected from areas on the substrates where the particle spacing was less than one particle diameter, or where the particles have aggregated. The results are interpreted in the context that the underlying physical phenomenon of MIL is a sequential two-photon absorption process, where the first event is driven by the plasmon resonance. It is evident that gold nanospheres in this size range have to be closely spaced or clustered to exhibit detectable MIL using far-field MPM operating in the NIR region.

  12. Stepwise multiphoton activation fluorescence reveals a new method of melanin detection

    Science.gov (United States)

    Lai, Zhenhua; Kerimo, Josef; Mega, Yair; DiMarzio, Charles A.

    2013-06-01

    The stepwise multiphoton activated fluorescence (SMPAF) of melanin, activated by a continuous-wave mode near infrared (NIR) laser, reveals a broad spectrum extending from the visible spectra to the NIR and has potential application for a low-cost, reliable method of detecting melanin. SMPAF images of melanin in mouse hair and skin are compared with conventional multiphoton fluorescence microscopy and confocal reflectance microscopy (CRM). By combining CRM with SMPAF, we can locate melanin reliably. However, we have the added benefit of eliminating background interference from other components inside mouse hair and skin. The melanin SMPAF signal from the mouse hair is a mixture of a two-photon process and a third-order process. The melanin SMPAF emission spectrum is activated by a 1505.9-nm laser light, and the resulting spectrum has a peak at 960 nm. The discovery of the emission peak may lead to a more energy-efficient method of background-free melanin detection with less photo-bleaching.

  13. PScan 1.0: flexible software framework for polygon based multiphoton microscopy

    Science.gov (United States)

    Li, Yongxiao; Lee, Woei Ming

    2016-12-01

    Multiphoton laser scanning microscopes exhibit highly localized nonlinear optical excitation and are powerful instruments for in-vivo deep tissue imaging. Customized multiphoton microscopy has a significantly superior performance for in-vivo imaging because of precise control over the scanning and detection system. To date, there have been several flexible software platforms catered to custom built microscopy systems i.e. ScanImage, HelioScan, MicroManager, that perform at imaging speeds of 30-100fps. In this paper, we describe a flexible software framework for high speed imaging systems capable of operating from 5 fps to 1600 fps. The software is based on the MATLAB image processing toolbox. It has the capability to communicate directly with a high performing imaging card (Matrox Solios eA/XA), thus retaining high speed acquisition. The program is also designed to communicate with LabVIEW and Fiji for instrument control and image processing. Pscan 1.0 can handle high imaging rates and contains sufficient flexibility for users to adapt to their high speed imaging systems.

  14. Multi-photon creation and single-photon annihilation of electron-positron pairs

    Energy Technology Data Exchange (ETDEWEB)

    Hu, Huayu

    2011-04-27

    In this thesis we study multi-photon e{sup +}e{sup -} pair production in a trident process, and singlephoton e{sup +}e{sup -} pair annihilation in a triple interaction. The pair production is considered in the collision of a relativistic electron with a strong laser beam, and calculated within the theory of laser-dressed quantum electrodynamics. A regularization method is developed systematically for the resonance problem arising in the multi-photon process. Total production rates, positron spectra, and relative contributions of different reaction channels are obtained in various interaction regimes. Our calculation shows good agreement with existing experimental data from SLAC, and adds further insights into the experimental findings. Besides, we study the process in a manifestly nonperturbative domain, whose accessibility to future all-optical experiments based on laser acceleration is shown. In the single-photon e{sup +}e{sup -} pair annihilation, the recoil momentum is absorbed by a spectator particle. Various kinematic configurations of the three incoming particles are examined. Under certain conditions, the emitted photon exhibits distinct angular and polarization distributions which could facilitate the detection of the process. Considering an equilibrium relativistic e{sup +}e{sup -} plasma, it is found that the single-photon process becomes the dominant annihilation channel for plasma temperatures above 3 MeV. Multi-particle correlation effects are therefore essential for the e{sup +}e{sup -} dynamics at very high density. (orig.)

  15. Multi-photon creation and single-photon annihilation of electron-positron pairs

    International Nuclear Information System (INIS)

    Hu, Huayu

    2011-01-01

    In this thesis we study multi-photon e + e - pair production in a trident process, and singlephoton e + e - pair annihilation in a triple interaction. The pair production is considered in the collision of a relativistic electron with a strong laser beam, and calculated within the theory of laser-dressed quantum electrodynamics. A regularization method is developed systematically for the resonance problem arising in the multi-photon process. Total production rates, positron spectra, and relative contributions of different reaction channels are obtained in various interaction regimes. Our calculation shows good agreement with existing experimental data from SLAC, and adds further insights into the experimental findings. Besides, we study the process in a manifestly nonperturbative domain, whose accessibility to future all-optical experiments based on laser acceleration is shown. In the single-photon e + e - pair annihilation, the recoil momentum is absorbed by a spectator particle. Various kinematic configurations of the three incoming particles are examined. Under certain conditions, the emitted photon exhibits distinct angular and polarization distributions which could facilitate the detection of the process. Considering an equilibrium relativistic e + e - plasma, it is found that the single-photon process becomes the dominant annihilation channel for plasma temperatures above 3 MeV. Multi-particle correlation effects are therefore essential for the e + e - dynamics at very high density. (orig.)

  16. Insights on proximity effect and multiphoton induced luminescence from gold nanospheres in far field optical microscopy

    International Nuclear Information System (INIS)

    Borglin, Johan; Guldbrand, Stina; Evenbratt, Hanne; Kirejev, Vladimir; Ericson, Marica B.; Grönbeck, Henrik

    2015-01-01

    Gold nanoparticles can be visualized in far-field multiphoton laser-scanning microscopy (MPM) based on the phenomena of multiphoton induced luminescence (MIL). This is of interest for biomedical applications, e.g., for cancer diagnostics, as MPM allows for working in the near-infrared (NIR) optical window of tissue. It is well known that the aggregation of particles causes a redshift of the plasmon resonance, but its implications for MIL applying far-field MPM should be further exploited. Here, we explore MIL from 10 nm gold nanospheres that are chemically deposited on glass substrates in controlled coverage gradients using MPM operating in NIR range. The substrates enable studies of MIL as a function of inter-particle distance and clustering. It was shown that MIL was only detected from areas on the substrates where the particle spacing was less than one particle diameter, or where the particles have aggregated. The results are interpreted in the context that the underlying physical phenomenon of MIL is a sequential two-photon absorption process, where the first event is driven by the plasmon resonance. It is evident that gold nanospheres in this size range have to be closely spaced or clustered to exhibit detectable MIL using far-field MPM operating in the NIR region

  17. Single photon and multiphoton events with missing energy in $e^{+} e^{-}$ collisions at LEP

    CERN Document Server

    Achard, P; Aguilar-Benítez, M; Alcaraz, J; Alemanni, G; Allaby, James V; Aloisio, A; Alviggi, M G; Anderhub, H; Andreev, V P; Anselmo, F; Arefev, A; Azemoon, T; Aziz, T; Bagnaia, P; Bajo, A; Baksay, G; Baksay, L; Baldew, S V; Banerjee, S; Banerjee, Sw; Barczyk, A; Barillère, R; Bartalini, P; Basile, M; Batalova, N; Battiston, R; Bay, A; Becattini, F; Becker, U; Behner, F; Bellucci, L; Berbeco, R; Berdugo, J; Berges, P; Bertucci, B; Betev, B L; Biasini, M; Biglietti, M; Biland, A; Blaising, J J; Blyth, S C; Bobbink, Gerjan J; Böhm, A; Boldizsar, L; Borgia, B; Bottai, S; Bourilkov, D; Bourquin, Maurice; Braccini, S; Branson, J G; Brochu, F; Burger, J D; Burger, W J; Cai, X D; Capell, M; Cara Romeo, G; Carlino, G; Cartacci, A M; Casaus, J; Cavallari, F; Cavallo, N; Cecchi, C; Cerrada, M; Chamizo-Llatas, M; Chang, Y H; Chemarin, M; Chen, A; Chen, G; Chen, G M; Chen, H F; Chen, H S; Chiefari, G; Cifarelli, Luisa; Cindolo, F; Clare, I; Clare, R; Coignet, G; Colino, N; Costantini, S; de la Cruz, B; Cucciarelli, S; van Dalen, J A; De Asmundis, R; Déglon, P L; Debreczeni, J; Degré, A; Dehmelt, K; Deiters, K; Della Volpe, D; Delmeire, E; Denes, P; De Notaristefani, F; De Salvo, A; Diemoz, M; Dierckxsens, M; Dionisi, C; Dittmar, M; Doria, A; Dova, M T; Duchesneau, D; Duda, M; Echenard, B; Eline, A; El-Hage, A; El-Mamouni, H; Engler, A; Eppling, F J; Extermann, P; Falagán, M A; Falciano, S; Favara, A; Fay, J; Fedin, O; Felcini, M; Ferguson, T; Fesefeldt, H S; Fiandrini, E; Field, J H; Filthaut, F; Fisher, P H; Fisher, W; Fisk, I; Forconi, G; Freudenreich, Klaus; Furetta, C; Galaktionov, Yu; Ganguli, S N; García-Abia, P; Gataullin, M; Gentile, S; Giagu, S; Gong, Z F; Grenier, G; Grimm, O; Grünewald, M W; Guida, M; van Gulik, R; Gupta, V K; Gurtu, A; Gutay, L J; Haas, D; Hatzifotiadou, D; Hebbeker, T; Hervé, A; Hirschfelder, J; Hofer, H; Hohlmann, M; Holzner, G; Hou, S R; Hu, Y; Jin, B N; Jones, L W; de Jong, P; Josa-Mutuberria, I; Käfer, D; Kaur, M; Kienzle-Focacci, M N; Kim, J K; Kirkby, Jasper; Kittel, E W; Klimentov, A; König, A C; Kopal, M; Koutsenko, V F; Kräber, M H; Krämer, R W; Krüger, A; Kunin, A; Ladrón de Guevara, P; Laktineh, I; Landi, G; Lebeau, M; Lebedev, A; Lebrun, P; Lecomte, P; Lecoq, P; Le Coultre, P; Le Goff, J M; Leiste, R; Levtchenko, M; Levchenko, P M; Li, C; Likhoded, S; Lin, C H; Lin, W T; Linde, Frank L; Lista, L; Liu, Z A; Lohmann, W; Longo, E; Lü, Y S; Luci, C; Luminari, L; Lustermann, W; Ma Wen Gan; Malgeri, L; Malinin, A; Maña, C; Mans, J; Martin, J P; Marzano, F; Mazumdar, K; McNeil, R R; Mele, S; Merola, L; Meschini, M; Metzger, W J; Mihul, A; Milcent, H; Mirabelli, G; Mnich, J; Mohanty, G B; Muanza, G S; Muijs, A J M; Musicar, B; Musy, M; Nagy, S; Natale, S; Napolitano, M; Nessi-Tedaldi, F; Newman, H; Nisati, A; Novák, T; Nowak, H; Ofierzynski, R A; Organtini, G; Pal, I; Palomares, C; Paolucci, P; Paramatti, R; Passaleva, G; Patricelli, S; Paul, T; Pauluzzi, M; Paus, C; Pauss, Felicitas; Pedace, M; Pensotti, S; Perret-Gallix, D; Petersen, B; Piccolo, D; Pierella, F; Pioppi, M; Piroué, P A; Pistolesi, E; Plyaskin, V; Pohl, M; Pozhidaev, V; Pothier, J; Prokofev, D; Prokofiev, D O; Quartieri, J; Rahal-Callot, G; Rahaman, M A; Raics, P; Raja, N; Ramelli, R; Rancoita, P G; Ranieri, R; Raspereza, A V; Razis, P A; Ren, D; Rescigno, M; Reucroft, S; Riemann, S; Riles, K; Roe, B P; Romero, L; Rosca, A; Rosenbleck, C; Rosier-Lees, S; Roth, S; Rubio, J A; Ruggiero, G; Rykaczewski, H; Sakharov, A; Saremi, S; Sarkar, S; Salicio, J; Sánchez, E; Schäfer, C; Shchegelskii, V; Schopper, Herwig Franz; Schotanus, D J; Sciacca, C; Servoli, L; Shevchenko, S; Shivarov, N; Shoutko, V; Shumilov, E; Shvorob, A V; Son, D; Souga, C; Spillantini, P; Steuer, M; Stickland, D P; Stoyanov, B; Strässner, A; Sudhakar, K; Sultanov, G G; Sun, L Z; Sushkov, S; Suter, H; Swain, J D; Szillási, Z; Tang, X W; Tarjan, P; Tauscher, Ludwig; Taylor, L; Tellili, B; Teyssier, D; Timmermans, C; Ting, Samuel C C; Ting, S M; Tonwar, S C; Tóth, J; Tully, C; Tung, K L; Ulbricht, J; Valente, E; Van de Walle, R T; Vásquez, R; Veszpremi, V; Vesztergombi, G; Vetlitskii, I; Vicinanza, D; Viertel, Gert M; Villa, S; Vivargent, M; Vlachos, S; Vodopyanov, I; Vogel, H; Vogt, H; Vorobev, I; Vorobyov, A A; Wadhwa, M; Wang, Q; Wang, X L; Wang, Z M; Weber, M; Wienemann, P; Wilkens, H; Wynhoff, S; Xia, L; Xu, Z Z; Yamamoto, J; Yang, B Z; Yang, C G; Yang, H J; Yang, M; Yeh, S C; Zalite, A; Zalite, Yu; Zhang, Z P; Zhao, J; Zhu, G Y; Zhu, R Y; Zhuang, H L; Zichichi, A; Zimmermann, B; Zöller, M

    2004-01-01

    Single- and multi-photon events with missing energy are selected in 619/pb of data collected by the L3 detector at LEP at centre-of-mass energies between 189GeV and 209GeV. The cross sections of the process e^+e^- -> nu nu gamma (gamma) are found to be in agreement with the Standard Model expectations, and the number of light neutrino species is determined, including lower energy data, to be N_nu = 2.98 +/- 0.05 +/- 0.04. Selection results are also given in the form of tables which can be used to test future models involving single- and multi-photon signatures at LEP. These final states are also predicted by models with large extra dimensions and by several supersymmetric models. No evidence for such models is found. Among others, lower limits between 1.5TeV and 0.65TeV are set, at 95% confidence level, on the new scale of gravity for the number of extra dimensions between 2 and 8.

  18. IR and visible luminescence studies in the infrared multiphoton dissociation of 1,2-dibromo-1,1-difluoroethane

    Science.gov (United States)

    Pushpa, K. K.; Kumar, Awadhesh; Vatsa, R. K.; Naik, P. D.; Annaji Rao, K.; Mittal, J. P.; Parthasarathy, V.; Sarkar, S. K.

    1995-07-01

    The infrared multiphoton dissociation of 1,2-dibromo-1,1-difluoroethane gives rise to IR and visible luminescence. Vibrationally excited parent molecules dissociate via two primary channels yielding bromine and vibrationally excited HBr. The strong visible emission observed between 350 to 750 nm has been assigned to electronically excited carbene CF 2Br CH.

  19. Comparison of analytical and Monte Carlo calculations of multi-photon effects in bremsstrahlung emission by high-energy electrons

    DEFF Research Database (Denmark)

    Mangiarotti, Alessio; Sona, Pietro; Ballestrero, Sergio

    2012-01-01

    Approximate analytical calculations of multi-photon effects in the spectrum of total radiated energy by high-energy electrons crossing thin targets are compared to the results of Monte Carlo type simulations. The limits of validity of the analytical expressions found in the literature are establi...

  20. In vivo 3D measurement of moxifloxacin and gatifloxacin distributions in the mouse cornea using multiphoton microscopy

    Science.gov (United States)

    Lee, Seunghun; Lee, Jun Ho; Park, Jin Hyoung; Yoon, Yeoreum; Chung, Wan Kyun; Tchah, Hungwon; Kim, Myoung Joon; Kim, Ki Hean

    2016-05-01

    Moxifloxacin and gatifloxacin are fourth-generation fluoroquinolone antibiotics used in the clinic to prevent or treat ocular infections. Their pharmacokinetics in the cornea is usually measured from extracted ocular fluids or tissues, and in vivo direct measurement is difficult. In this study multiphoton microscopy (MPM), which is a 3D optical microscopic technique based on multiphoton fluorescence, was applied to the measurement of moxifloxacin and gatifloxacin distribution in the cornea. Intrinsic multiphoton fluorescence properties of moxifloxacin and gatifloxacin were characterized, and their distributions in mouse cornea in vivo were measured by 3D MPM imaging. Both moxifloxacin and gatifloxacin had similar multiphoton spectra, while moxifloxacin had stronger fluorescence than gatifloxacin. MPM imaging of mouse cornea in vivo showed (1) moxifloxacin had good penetration through the superficial corneal epithelium, while gatifloxacin had relatively poor penetration, (2) both ophthalmic solutions had high intracellular distribution. In vivo MPM results were consistent with previous studies. This study demonstrates the feasibility of MPM as a method for in vivo direct measurement of moxifloxacin and gatifloxacin in the cornea.

  1. Photoleucine Survives Backbone Cleavage by Electron Transfer Dissociation. A Near-UV Photodissociation and Infrared Multiphoton Dissociation Action Spectroscopy Study

    Czech Academy of Sciences Publication Activity Database

    Shaffer, C. J.; Martens, J.; Marek, Aleš; Oomens, J.; Tureček, F.

    2016-01-01

    Roč. 27, č. 7 (2016), s. 1176-1185 ISSN 1044-0305 Institutional support: RVO:61388963 Keywords : peptide ions * electron transfer dissociation * photoleucine label * near-UV photodissociation * infrared multiphoton dissociation action spectroscopy Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.786, year: 2016

  2. Generation of a multi-photon Greenberger-Horne-Zeilinger state with linear optical elements and photon detectors

    International Nuclear Information System (INIS)

    Zou, X B; Pahlke, K; Mathis, W

    2005-01-01

    We present a scheme to generate a multi-photon Greenberger-Horne-Zeilinger (GHZ) state by using single-photon sources, linear optical elements and photon detectors. Such a maximum entanglement has wide applications in the demonstration of quantum nonlocality and quantum information processing

  3. Resonance-enhanced multiphoton ionization photoelectron spectroscopy of even-parity autoionizing Rydberg states of atomic sulphur

    NARCIS (Netherlands)

    Woutersen, S.; de Milan, J.B.; de Lange, C.A.; Buma, W.J.

    1997-01-01

    Several previously unobserved Rydberg states of the sulphur atom above the lowest ionization threshold are identified and assigned using (2 + 1) resonance-enhanced multiphoton-ionization photoelectron spectroscopy. All states were accessed by two-photon transitions from either the 3P ground or the

  4. Multi-photon ionization of atoms in intense short-wavelength radiation fields

    Science.gov (United States)

    Meyer, Michael

    2015-05-01

    The unprecedented characteristics of XUV and X-ray Free Electron Lasers (FELs) have stimulated numerous investigations focusing on the detailed understanding of fundamental photon-matter interactions in atoms and molecules. In particular, the high intensities (up to 106 W/cm2) giving rise to non-linear phenomena in the short wavelength regime. The basic phenomenology involves the production of highly charged ions via electron emission to which both sequential and direct multi-photon absorption processes contribute. The detailed investigation of the role and relative weight of these processes under different conditions (wavelength, pulse duration, intensity) is the key element for a comprehensive understanding of the ionization dynamics. Here the results of recent investigations are presented, performed at the FELs in Hamburg (FLASH) and Trieste (FERMI) on atomic systems with electronic structures of increasing complexity (Ar, Ne and Xe). Mainly, electron spectroscopy is used to obtain quantitative information about the relevance of various multi-photon ionization processes. For the case of Ar, a variety of processes including above threshold ionization (ATI) from 3p and 3s valence shells, direct 2p two-photon ionization and resonant 2p-4p two-photon excitations were observed and their role was quantitatively determined comparing the experimental ionization yields to ab-initio calculations of the cross sections for the multi-photon processes. Using Ar as a benchmark to prove the reliability of the combined experimental and theoretical approach, the more complex and intriguing case of Xe was studied. Especially, the analysis of the two-photon ATI from the Xe 4d shell reveals new insight into the character of the 4d giant resonance, which was unresolved in the linear one-photon regime. Finally, the influence of intense XUV radiation to the relaxation dynamics of the Ne 2s-3p resonance was investigated by angle-resolved electron spectroscopy, especially be observing

  5. Coherent beam control through inhomogeneous media in multi-photon microscopy

    Science.gov (United States)

    Paudel, Hari Prasad

    Multi-photon fluorescence microscopy has become a primary tool for high-resolution deep tissue imaging because of its sensitivity to ballistic excitation photons in comparison to scattered excitation photons. The imaging depth of multi-photon microscopes in tissue imaging is limited primarily by background fluorescence that is generated by scattered light due to the random fluctuations in refractive index inside the media, and by reduced intensity in the ballistic focal volume due to aberrations within the tissue and at its interface. We built two multi-photon adaptive optics (AO) correction systems, one for combating scattering and aberration problems, and another for compensating interface aberrations. For scattering correction a MEMS segmented deformable mirror (SDM) was inserted at a plane conjugate to the objective back-pupil plane. The SDM can pre-compensate for light scattering by coherent combination of the scattered light to make an apparent focus even at a depths where negligible ballistic light remains (i.e. ballistic limit). This problem was approached by investigating the spatial and temporal focusing characteristics of a broad-band light source through strongly scattering media. A new model was developed for coherent focus enhancement through or inside the strongly media based on the initial speckle contrast. A layer of fluorescent beads under a mouse skull was imaged using an iterative coherent beam control method in the prototype two-photon microscope to demonstrate the technique. We also adapted an AO correction system to an existing in three-photon microscope in a collaborator lab at Cornell University. In the second AO correction approach a continuous deformable mirror (CDM) is placed at a plane conjugate to the plane of an interface aberration. We demonstrated that this "Conjugate AO" technique yields a large field-of-view (FOV) advantage in comparison to Pupil AO. Further, we showed that the extended FOV in conjugate AO is maintained over a

  6. Imaging photoelectron circular dichroism of chiral molecules by femtosecond multiphoton coincidence detection

    Energy Technology Data Exchange (ETDEWEB)

    Lehmann, C. Stefan; Ram, N. Bhargava; Janssen, Maurice H. M., E-mail: m.h.m.janssen@vu.nl [LaserLaB Amsterdam, VU University Amsterdam, De Boelelaan 1081, 1081 HV Amsterdam (Netherlands); Powis, Ivan [School of Chemistry, University of Nottingham, Nottingham NG7 2RD (United Kingdom)

    2013-12-21

    Here, we provide a detailed account of novel experiments employing electron-ion coincidence imaging to discriminate chiral molecules. The full three-dimensional angular scattering distribution of electrons is measured after photoexcitation with either left or right circular polarized light. The experiment is performed using a simplified photoelectron-photoion coincidence imaging setup employing only a single particle imaging detector. Results are reported applying this technique to enantiomers of the chiral molecule camphor after three-photon ionization by circularly polarized femtosecond laser pulses at 400 nm and 380 nm. The electron-ion coincidence imaging provides the photoelectron spectrum of mass-selected ions that are observed in the time-of-flight mass spectra. The coincident photoelectron spectra of the parent camphor ion and the various fragment ions are the same, so it can be concluded that fragmentation of camphor happens after ionization. We discuss the forward-backward asymmetry in the photoelectron angular distribution which is expressed in Legendre polynomials with moments up to order six. Furthermore, we present a method, similar to one-photon electron circular dichroism, to quantify the strength of the chiral electron asymmetry in a single parameter. The circular dichroism in the photoelectron angular distribution of camphor is measured to be 8% at 400 nm. The electron circular dichroism using femtosecond multiphoton excitation is of opposite sign and about 60% larger than the electron dichroism observed before in near-threshold one-photon ionization with synchrotron excitation. We interpret our multiphoton ionization as being resonant at the two-photon level with the 3s and 3p Rydberg states of camphor. Theoretical calculations are presented that model the photoelectron angular distribution from a prealigned camphor molecule using density functional theory and continuum multiple scattering X alpha photoelectron scattering calculations

  7. Modified method for intravital-fluorescent microscopy for on-line measurement of vasomotion in cutaneous microcirculation of mice in vivo

    International Nuclear Information System (INIS)

    Traikov, L.; Jia, F.; Ushiyama, A.; Masuda, H.; Ohkubo, C.

    2004-01-01

    The purpose of this study was establishing of the suitable method for long term investigations of changes of the blood vessel diameter up to 60 min continuous measurements with the main aim to be detected possible changes in low frequency component of vasomotion at cutaneous microcirculation of mice in vivo without using anesthetized animals. For investigation of micro-circulatory changes of blood vessel diameter, it is necessary to be used as high as possible quality video images with high resolution (1591x1061 pixels) - at this time the best method for this purpose is intravital microscopy, with high magnification and high resolution this method allows to be observed blood vessels from 5 to 100 μm - and also to be determined blood vessel wall and surrounded blood vessel cells. In this investigation BALB/c mice were used, and a Dorsal Skin-fold Chamber (DSC), were surgically implanted 4 days before the experiment. Intravital video-microscopy measurements of vasomotion in vivo were realized by 3CCD-camera, SONY Inc. and Nikon Inc. microscopy system. Fluorescence signal from the blood vessel, were enhanced by using DVS-3000 system (Hamamatsu Inc.) Vasomotion or temporal changes of the blood vessel diameter was measured by High-speed Digital Machine Vision System CV-2100 (KEYENCE Inc.), using an edge-gap detection algorithm (with a sampling time 389 ms), for on-line calculating the outer diameter of the blood vessels by means of fluorescence-image visualization (with precision 0.5 μm), after caudal vein injection of (from 50h150 μl per 25g animal) Fluorescein Isothiocyanate (FITC)-labeled Dextrans from 60-250 kDa concentration 2.5% in PBS solution pH=7.5. First part of this study was set up the intensity of the fluorescence using different concentrations and molecular weight Dextrans for visualization of the blood plasma with enough high intensity and as low as possible effects on the blood viscosity. Set up of the edge finding algorithm from off-line calculations

  8. Beyond the Floquet theorem: generalized Floquet formalisms and quasienergy methods for atomic and molecular multiphoton processes in intense laser fields

    Science.gov (United States)

    Chu, Shih-I.; Telnov, Dmitry A.

    2004-02-01

    The advancement of high-power and short-pulse laser technology in the past two decades has generated considerable interest in the study of multiphoton and very high-order nonlinear optical processes of atomic and molecular systems in intense and superintense laser fields, leading to the discovery of a host of novel strong-field phenomena which cannot be understood by the conventional perturbation theory. The Floquet theorem and the time-independent Floquet Hamiltonian method are powerful theoretical framework for the study of bound-bound multiphoton transitions driven by periodically time-dependent fields. However, there are a number of significant strong-field processes cannot be directly treated by the conventional Floquet methods. In this review article, we discuss several recent developments of generalized Floquet theorems, formalisms, and quasienergy methods, beyond the conventional Floquet theorem, for accurate nonperturbative treatment of a broad range of strong-field atomic and molecular processes and phenomena of current interests. Topics covered include (a) artificial intelligence (AI)-most-probable-path approach (MPPA) for effective treatment of ultralarge Floquet matrix problem; (b) non-Hermitian Floquet formalisms and complex quasienergy methods for nonperturbative treatment of bound-free and free-free processes such as multiphoton ionization (MPI) and above-threshold ionization (ATI) of atoms and molecules, multiphoton dissociation (MPD) and above-threshold dissociation (ATD) of molecules, chemical bond softening and hardening, charge-resonance enhanced ionization (CREI) of molecular ions, and multiple high-order harmonic generation (HHG), etc.; (c) many-mode Floquet theorem (MMFT) for exact treatment of multiphoton processes in multi-color laser fields with nonperiodic time-dependent Hamiltonian; (d) Floquet-Liouville supermatrix (FLSM) formalism for exact nonperturbative treatment of time-dependent Liouville equation (allowing for relaxations and

  9. Beyond the Floquet theorem: generalized Floquet formalisms and quasienergy methods for atomic and molecular multiphoton processes in intense laser fields

    International Nuclear Information System (INIS)

    Chu, S.-I.; Telnov, D.A.

    2004-01-01

    The advancement of high-power and short-pulse laser technology in the past two decades has generated considerable interest in the study of multiphoton and very high-order nonlinear optical processes of atomic and molecular systems in intense and superintense laser fields, leading to the discovery of a host of novel strong-field phenomena which cannot be understood by the conventional perturbation theory. The Floquet theorem and the time-independent Floquet Hamiltonian method are powerful theoretical framework for the study of bound-bound multiphoton transitions driven by periodically time-dependent fields. However, there are a number of significant strong-field processes cannot be directly treated by the conventional Floquet methods. In this review article, we discuss several recent developments of generalized Floquet theorems, formalisms, and quasienergy methods, beyond the conventional Floquet theorem, for accurate nonperturbative treatment of a broad range of strong-field atomic and molecular processes and phenomena of current interests. Topics covered include (a) artificial intelligence (AI)-most-probable-path approach (MPPA) for effective treatment of ultralarge Floquet matrix problem; (b) non-Hermitian Floquet formalisms and complex quasienergy methods for nonperturbative treatment of bound-free and free-free processes such as multiphoton ionization (MPI) and above-threshold ionization (ATI) of atoms and molecules, multiphoton dissociation (MPD) and above-threshold dissociation (ATD) of molecules, chemical bond softening and hardening, charge-resonance enhanced ionization (CREI) of molecular ions, and multiple high-order harmonic generation (HHG), etc.; (c) many-mode Floquet theorem (MMFT) for exact treatment of multiphoton processes in multi-color laser fields with nonperiodic time-dependent Hamiltonian; (d) Floquet-Liouville supermatrix (FLSM) formalism for exact nonperturbative treatment of time-dependent Liouville equation (allowing for relaxations and

  10. Visualization of dermal alteration in skin lesions with discoid lupus erythematosus by multiphoton microscopy

    International Nuclear Information System (INIS)

    Lin, L H; Yu, H B; Zhu, X Q; Zhuo, S M; Chen, J X; Wang, Y Y; Yang, Y H

    2013-01-01

    Discoid lupus erythematosus (DLE) is a chronic dermatological disease which lacks valid methods for early diagnosis and therapeutic monitoring. Considering the collagen and elastin disorder due to mucin deposition of DLE, multiphoton microscopy (MPM) imaging techniques were employed to obtain high-resolution collagen and elastin images from the dermis. The content and distribution of collagen and elastin were quantified to characterize the dermal pathological status of skin lesions with DLE in comparison with normal skin. Our results showed a significant difference between skin lesions with DLE and normal skin in terms of the morphological structure of collagen and elastin in the dermis, demonstrating the possibility of MPM for noninvasively tracking the pathological process of DLE even in its early stages and evaluating the therapeutic efficacy at the molecular level. (paper)

  11. Multiphoton effects in electron-ion scattering: A limitation of the cross-section treatment

    International Nuclear Information System (INIS)

    Torres Silva, H.; Sakanaka, P.H.; Braga, L.C.

    1991-07-01

    The differential cross-section for inelastic scattering in the presence of an intense laser field, when applied to the calculation of energy balance and heating by multiphoton process, is a problem which is not completely solved yet. One of the main difficulties is the calculation of the absorption coefficients α-bar for a monoenergetic beam of electrons scattered by a static potential. There are contradictory results shown by different authors. Here we have derived α-bar starting under the framework of quantum mechanics and then making the classical correspondence (h/2π → 0) according to the kinetic theory, and show that the absorption coefficient is always positive for all values of the particle incoming velocity, v-vector i . Furthermore, we show that in the calculation of α-bar we recover the Coulomb logarithm term. (author). 18 refs, 5 figs, 2 tabs

  12. Electron spectroscopy of He and NO using electron impact and multiphoton ionisation

    International Nuclear Information System (INIS)

    Kimman, J.T.N.

    1984-01-01

    This thesis describes two experimental studies which are intended to contribute to our knowledge of the structure of molecules and the decay dynamics of excited molecular states. The two studies have in common that they are both concerned with ionisation processes, in which an accurately known amount of energy is transferred to the target, and energy analysis of the ejected electrons is made. Ionisation is caused either by scattering electrons off the molecules (chapter 2: electron impact ionisation) or by a simultaneous absorption of several photons (chapter 3: multiphoton ionisation). In chapter 2 an electron impact ionisation experiment on Helium is described in which the kinematics of both the scattered and the ejected electrons is fully determined ((e,2e) experiment). (Auth.)

  13. Monitoring the effect of mechanical stress on mesenchymal stem cell collagen production by multiphoton microscopy

    Science.gov (United States)

    Chen, Wei-Liang; Chang, Chia-Cheng; Chiou, Ling-Ling; Li, Tsung-Hsien; Liu, Yuan; Lee, Hsuan-Shu; Dong, Chen-Yuan

    2008-02-01

    Tissue engineering is emerging as a promising method for repairing damaged tissues. Due to cartilage's common wear and injury, in vitro production of cartilage replacements have been an active area of research. Finding the optimal condition for the generation of the collagen matrix is crucial in reproducing cartilages that closely match those found in human. Using multiphoton autofluorescence and second-harmonic generation (SHG) microscopy we monitored the effect of mechanical stress on mesenchymal stem cell collagen production. Bone marrow mesenchymal stem cells in the form of pellets were cultured and periodically placed under different mechanical stress by centrifugation over a period of four weeks. The differently stressed samples were imaged several times during the four week period, and the collagen production under different mechanical stress is characterized.

  14. Non-dipole effects in multiphoton ionization of hydrogen atom in short superintense laser fields

    Energy Technology Data Exchange (ETDEWEB)

    Jobunga, Eric O. [AG Moderne Optik, Institut fuer Physik, Humboldt-Universitaet zu Berlin, Newtonstr. 15, D-12489 Berlin (Germany); Department of Mathematics and Physics, Technical University of Mombasa, P. O. Box 90420-80100, Mombasa (Kenya); Saenz, Alejandro [AG Moderne Optik, Institut fuer Physik, Humboldt-Universitaet zu Berlin, Newtonstr. 15, D-12489 Berlin (Germany)

    2014-07-01

    The development of novel light sources has enabled the realization of high-precision experiments investigating various non-linear processes in the dynamics of atomic, molecular, and ionic systems interacting with high intense laser pulses. At high intensities or short wavelengths, the analysis of these experiments would definitely require a reliable non-perturbative solution of the time-dependent Schroedinger or Dirac equation. These solutions should consider both the temporal and the spatial intensity variations of the laser pulse.We have solved the non-relativistic time dependent Schroedinger equation for a ground state hydrogen atom interacting with short intense spatially and temporally resolved laser fields corresponding to the multiphoton ATI regime for a monochromatic source with λ= 800 nm. We shall analyse the effects of the A{sup 2} term and the corresponding orders of the multipolar expansion of the transition matrix.

  15. The Application of Resonance-Enhanced Multiphoton Ionization Technique in Gas Chromatography Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Adan Li

    2014-01-01

    Full Text Available Gas chromatography resonance-enhanced multiphoton ionization time-of-flight mass spectrometry (GC/REMPI-TOFMS using a nanosecond laser has been applied to analyze the 16 polycyclic aromatic hydrocarbons (PAHs. The excited-state lifetime, absorption characters, and energy of electronic states of the 16 PAHs were investigated to optimize the ionization yield. A river water sample pretreated by means of solid phase extraction was analyzed to evaluate the performance of the analytical instrument. The results suggested that REMPI is superior to electron impact ionization method for soft ionization and suppresses the background signal due to aliphatic hydrocarbons. Thus, GC/REMPI-TOFMS is a more reliable method for the determination of PAHs present in the environment.

  16. Multiphoton microscopy guides neurotrophin modification with poly(ethylene glycol) to enhance interstitial diffusion

    Science.gov (United States)

    Stroh, Mark; Zipfel, Warren R.; Williams, Rebecca M.; Ma, Shu Chin; Webb, Watt W.; Saltzman, W. Mark

    2004-07-01

    Brain-derived neurotrophic factor (BDNF) is a promising therapeutic agent for the treatment of neurodegenerative diseases. However, the limited distribution of this molecule after administration into the brain tissue considerably hampers its efficacy. Here, we show how multiphoton microscopy of fluorescently tagged BDNF in brain-tissue slices provides a useful and rapid screening method for examining the diffusion of large molecules in tissues, and for studying the effects of chemical modifications-for example, conjugating with polyethylene glycol (PEG)-on the diffusion constant. This single variable, obtained by monitoring short-term diffusion in real time, can be effectively used for rational drug design. In this study on fluorescently tagged BDNF and BDNF-PEG, we identify slow diffusion as a major contributing factor to the limited penetration of BDNF, and demonstrate how chemical modification can be used to overcome this barrier.

  17. MOLECULAR BEAM STUDIES OF IR LASER INDUCED MULTIPHOTON DISSOCIATION AND VIBRATIONAL PREDISSOCIATION

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Yuan T.; Shen, Y. Ron

    1980-06-01

    The advancement of crossed molecular beam methods, modern spectroscopy and laser technology allows us to observe chemical reactions on atomic and molecular levels in great detail. After a brief history of crossed molecular beams studies, the author describes and discusses the universal molecular beam apparatus and gives examples of crossed molecular beam studies. The crossed beam technique is compared to other techniques used to provide microscopic information on reaction dynamics. Application of crossed laser and molecular beam studies to the problem of IR multiphoton dissociation of polyatomic molecules is discussed. Study of vibrational predissociation of hydrogen-bonded and van der Waals molecular clusters are discussed. Future cases that the author considers worth pursuing that could benefit from the collisionless environment of molecular beams are enumerated.

  18. Time and spectrum-resolving multiphoton correlator for 300–900 nm

    Energy Technology Data Exchange (ETDEWEB)

    Johnsen, Kelsey D.; Thibault, Marilyne; Jennewein, Thomas [Institute for Quantum Computing and Department for Physics and Astronomy, University of Waterloo, 200 University Ave. West, Waterloo, Ontario N2L 3G1 (Canada); Kolenderski, Piotr, E-mail: kolenderski@fizyka.umk.pl [Institute for Quantum Computing and Department for Physics and Astronomy, University of Waterloo, 200 University Ave. West, Waterloo, Ontario N2L 3G1 (Canada); Faculty of Physics, Astronomy and Informatics, Institute of Physics, Nicolaus Copernicus University, Grudziadzka 5, 87-100 Torun (Poland); Scarcella, Carmelo; Tosi, Alberto [Dipartimento di Elettronica, Informazione e Bioingegneria, Politecnico di Milano, Piazza Leonardo da Vinci 32, I-20133 Milano (Italy)

    2014-10-14

    We demonstrate a single-photon sensitive spectrometer in the visible range, which allows us to perform time-resolved and multi-photon spectral correlation measurements at room temperature. It is based on a monochromator composed of two gratings, collimation optics, and an array of single photon avalanche diodes. The time resolution can reach 110 ps and the spectral resolution is 2 nm/pixel, limited by the design of the monochromator. This technique can easily be combined with commercial monochromators and can be useful for joint spectrum measurements of two photons emitted in the process of parametric down conversion, as well as time-resolved spectrum measurements in optical coherence tomography or medical physics applications.

  19. Sub-15 fs multiphoton lithography of three-dimensional structures for live cell applications

    International Nuclear Information System (INIS)

    Licht, Martin; Uchugonova, Aisada; König, Karsten; Straub, Martin

    2012-01-01

    Development, morphology and intratissue location of cells are influenced by the 3D nano- and microenvironment. In this paper we demonstrate multiphoton photopolymerization to generate three-dimensional structures for cell culture applications with micro- and nanotopographic features using SU-8 photoresist and mr-NIL 6000 nanoimprint resist. Moving the focal spot of high-repetition rate near-infrared sub-15 fs pulsed laser light by a galvanometric beam scanner in combination with a piezoelectric vertical stage, nearly arbitrary trajectories of polymerized photoresist were generated. This technique can be used to generate cage structures with submicron interior features for live cell applications. Preliminary experiments with PC-3 and HT-1080 cells indicate the influence of the structures on cell behavior. (paper)

  20. Structural and dynamical aspects of skin studied by multiphoton excitation fluorescence microscopy-based methods

    DEFF Research Database (Denmark)

    Bloksgaard, Maria; Brewer, Jonathan R.; Bagatolli, Luis

    2013-01-01

    ' parameters. Specifically, by applying these methods, spatially resolved maps of water dipolar relaxation (generalized polarization function using the 6-lauroyl-2-(N,N-dimethylamino)naphthale probe), activity of protons (fluorescence lifetime imaging using a proton sensitive fluorescence probe--2,7-bis-(2......-carboxyethyl)-5-(and-6)-carboxyfluorescein) and diffusion coefficients of distinct fluorescence probes (raster imaging correlation spectroscopy) can be obtained from different regions of the tissue. Comparative studies of different tissue strata, but also between equivalent regions of normal and abnormal......This mini-review reports on applications of particular multiphoton excitation microscopy-based methodologies employed in our laboratory to study skin. These approaches allow in-depth optical sectioning of the tissue, providing spatially resolved information on specific fluorescence probes...

  1. Simultaneous multiphoton processes in the interaction of atoms with electromagnetic fields

    International Nuclear Information System (INIS)

    Levine, A.M.; Schreiber, W.M.; Weiszmann, A.N.

    1984-01-01

    It is impossible to obtain an exact description of multiphoton processes in the interaction of electromagnetic fields with atomic systems. Approximate approaches must be used to describe the physically different effects that can occur. One effect is the stepwise absorption/emission of many photons by a N-level system that evolves dynamically in between each absorption/emission. Another effect is described in the theories of Raman processes where the simultaneous absorption/emission of many photons is considered. In this paper, consideration is given to both processes allowing interference between the stepwise and simultaneous absorptions. An approximate Hamiltonian is obtained from the quantum mechanical multipole expansion. An exact solution of an atom-field system subject to this Hamiltonian will be presented. The extension of the method to multiple electromagnetic fields is discussed

  2. Differentiating fibroadenoma and ductal carcinoma in situ from normal breast tissue by multiphoton microscopy

    Science.gov (United States)

    Nie, Yuting; Wu, Yan; Lian, Yuane; Fu, Fangmeng; Wang, Chuan; Chen, Jianxin

    2014-09-01

    Fibroadenoma (FA) is the most common benign tumor of the female breast and several studies have reported that women with it have increased risk of breast cancer. While the ductal carcinoma in situ (DCIS) is a very early form of breast cancer. Thus, early detections of FA and DCIS are critical for improving breast tumor outcome and survival. In this paper, we use multiphoton microscopy (MPM) to obtain the high-contrast images of fresh, unfixed, unstained human breast specimens (normal breast tissue, FA and DCIS). Our results show that MPM has the ability to identify the characteristics of FA and DCIS including changes of duct architecture and collagen morphology. These results are consistent with the histological results. With the advancement of MPM, the technique has potential ability to serve as a real-time noninvasive imaging tool for early detection of breast tumor.

  3. A multiphoton laser scanning microscope setup for transcranial in vivo brain imaging on mice

    Science.gov (United States)

    Nase, Gabriele; Helm, P. Johannes; Reppen, Trond; Ottersen, Ole Petter

    2005-12-01

    We describe a multiphoton laser scanning microscope setup for transcranial in vivo brain imaging in mice. The modular system is based on a modified industrial standard Confocal Scanning Laser Microscope (CSLM) and is assembled mainly from commercially available components. A special multifunctional stage, which is optimized for both laser scanning microscopic observation and preparative animal surgery, has been developed and built. The detection unit includes a highly efficient photomultiplier tube installed in a Peltier-cooled thermal box shielding the detector from changes in room temperature and from distortions caused by external electromagnetic fields. The images are recorded using a 12-bit analog-to-digital converter. Depending on the characteristics of the staining, individual nerve cells can be imaged down to at least 100μm below the intact cranium and down to at least 200μm below the opened cranium.

  4. Pancreatic cancer cell detection by targeted lipid microbubbles and multiphoton imaging

    Science.gov (United States)

    Cromey, Benjamin; McDaniel, Ashley; Matsunaga, Terry; Vagner, Josef; Kieu, Khanh Quoc; Banerjee, Bhaskar

    2018-04-01

    Surgical resection of pancreatic cancer represents the only chance of cure and long-term survival in this common disease. Unfortunately, determination of a cancer-free margin at surgery is based on one or two tiny frozen section biopsies, which is far from ideal. Not surprisingly, cancer is usually left behind and is responsible for metastatic disease. We demonstrate a method of receptor-targeted imaging using peptide ligands, lipid microbubbles, and multiphoton microscopy that could lead to a fast and accurate way of examining the entire cut surface during surgery. Using a plectin-targeted microbubble, we performed a blinded in-vitro study to demonstrate avid binding of targeted microbubbles to pancreatic cancer cells but not noncancerous cell lines. Further work should lead to a much-needed point-of-care diagnostic test for determining clean margins in oncologic surgery.

  5. Relativistic theory of tunnel and multiphoton ionization of atoms in a strong laser field

    International Nuclear Information System (INIS)

    Popov, V. S.; Karnakov, B. M.; Mur, V. D.; Pozdnyakov, S. G.

    2006-01-01

    Relativistic generalization is developed for the semiclassical theory of tunnel and multiphoton ionization of atoms and ions in the field of an intense electromagnetic wave (Keldysh theory). The cases of linear, circular, and elliptic polarizations of radiation are considered. For arbitrary values of the adiabaticity parameter γ, the exponential factor in the ionization rate for a relativistic bound state is calculated. For low-frequency laser radiation , an asymptotically exact formula for the tunnel ionization rate for the atomic s level is obtained including the Coulomb, spin, and adiabatic corrections and the preexponential factor. The ionization rate for the ground level of a hydrogen-like atom (ion) with Z ≤ 100 is calculated as a function of the laser radiation intensity. The range of applicability is determined for nonrelativistic ionization theory. The imaginary time method is used in the calculations

  6. Field enhancement of multiphoton induced luminescence processes in ZnO nanorods

    Science.gov (United States)

    Hyyti, Janne; Perestjuk, Marko; Mahler, Felix; Grunwald, Rüdiger; Güell, Frank; Gray, Ciarán; McGlynn, Enda; Steinmeyer, Günter

    2018-03-01

    The near-ultraviolet photoluminescence of ZnO nanorods induced by multiphoton absorption of unamplified Ti:sapphire pulses is investigated. Power dependence measurements have been conducted with an adaptation of the ultrashort pulse characterization method of interferometric frequency-resolved optical gating. These measurements enable the separation of second harmonic and photoluminescence bands due to their distinct coherence properties. A detailed analysis yields fractional power dependence exponents in the range of 3-4, indicating the presence of multiple nonlinear processes. The range in measured exponents is attributed to differences in local field enhancement, which is supported by independent photoluminescence and structural measurements. Simulations based on Keldysh theory suggest contributions by three- and four-photon absorption as well as avalanche ionization in agreement with experimental findings.

  7. 2008 Multiphoton Processes Gordon Research Conferences - June 8-13, 2008

    Energy Technology Data Exchange (ETDEWEB)

    Mette B. Gaarde

    2009-08-28

    In 2008 the Gordon Research Conference on Multiphoton Processes is held for the 14th time. The meeting continues to evolve as it embraces both the rapid technological and intellectual growth in the field as well as the multi-disciplinary expertise of the participants. This time the sessions will focus on: (1) Attosecond Science; (2) Free-electron laser experiments and theory; (3) Ultrafast dynamics of molecules; (4) Laser control of molecules; (5) Ultrafast imaging; (6) Super-high intensity and x-rays; (7) Strong field processes in molecules; and (8) Control atoms with light and vice versa. The scientific program will blur traditional disciplinary boundaries as the presenters and discussion leaders involve chemists, physicists, and optical engineers, representing both experiment and theory. The broad range of expertise and different perspectives of attendees should provide a stimulating and unique environment for solving problems and developing new ideas in this rapidly evolving field.

  8. Multiphoton photoemission from a copper cathode illuminated by ultrashort laser pulses in an RF photoinjector.

    Science.gov (United States)

    Musumeci, P; Cultrera, L; Ferrario, M; Filippetto, D; Gatti, G; Gutierrez, M S; Moody, J T; Moore, N; Rosenzweig, J B; Scoby, C M; Travish, G; Vicario, C

    2010-02-26

    In this Letter we report on the use of ultrashort infrared laser pulses to generate a copious amount of electrons by a copper cathode in an rf photoinjector. The charge yield verifies the generalized Fowler-Dubridge theory for multiphoton photoemission. The emission is verified to be prompt using a two pulse autocorrelation technique. The thermal emittance associated with the excess kinetic energy from the emission process is comparable with the one measured using frequency tripled uv laser pulses. In the high field of the rf gun, up to 50 pC of charge can be extracted from the cathode using a 80 fs long, 2 microJ, 800 nm pulse focused to a 140 mum rms spot size. Taking into account the efficiency of harmonic conversion, illuminating a cathode directly with ir laser pulses can be the most efficient way to employ the available laser power.

  9. Multiphoton Photoemission from a Copper Cathode Illuminated by Ultrashort Laser Pulses in an rf Photoinjector

    International Nuclear Information System (INIS)

    Musumeci, P.; Gutierrez, M. S.; Moody, J. T.; Moore, N.; Rosenzweig, J. B.; Scoby, C. M.; Travish, G.; Cultrera, L.; Ferrario, M.; Filippetto, D.; Gatti, G.; Vicario, C.

    2010-01-01

    In this Letter we report on the use of ultrashort infrared laser pulses to generate a copious amount of electrons by a copper cathode in an rf photoinjector. The charge yield verifies the generalized Fowler-Dubridge theory for multiphoton photoemission. The emission is verified to be prompt using a two pulse autocorrelation technique. The thermal emittance associated with the excess kinetic energy from the emission process is comparable with the one measured using frequency tripled uv laser pulses. In the high field of the rf gun, up to 50 pC of charge can be extracted from the cathode using a 80 fs long, 2 μJ, 800 nm pulse focused to a 140 μm rms spot size. Taking into account the efficiency of harmonic conversion, illuminating a cathode directly with ir laser pulses can be the most efficient way to employ the available laser power.

  10. Monitoring the beam flux in molecular beam epitaxy using laser multiphoton ionization

    International Nuclear Information System (INIS)

    Chien, R.; Sogard, M.R.

    1990-01-01

    In this paper, we will describe a method using laser nonresonant multiphoton ionization to measure beam flux in molecular beam epitaxy (MBE) systems. The results were obtained in a test chamber where a focused excimer laser beam was used to photoionize a small fraction of the atomic and molecular beams. The constituents of the beams were identified by a time-of-flight mass spectrometer. Ion signal strength was found to be directly correlated to the temperature of the atomic beam oven. Good stability and sensitivity on gallium, aluminum, and silicon atomic beams was demonstrated. Arsenic was also detected. We demonstrated very sensitive detection of contaminant atomic and molecular constituents of our system. We have also detected the presence of short-term fluctuations in the gallium flux from an effusion source. These fluctuations, previously suspected, can be in excess of ±10%

  11. Coherent Control of Multiphoton Transitions in the Gas and Condensed Phases with Shaped Ultrashort Pulses

    International Nuclear Information System (INIS)

    Dantus, Marcos

    2008-01-01

    Controlling laser-molecule interactions has become an integral part of developing devices and applications in spectroscopy, microscopy, optical switching, micromachining and photochemistry. Coherent control of multiphoton transitions could bring a significant improvement of these methods. In microscopy, multi-photon transitions are used to activate different contrast agents and suppress background fluorescence; coherent control could generate selective probe excitation. In photochemistry, different dissociative states are accessed through two, three, or more photon transitions; coherent control could be used to select the reaction pathway and therefore the yield-specific products. For micromachining and processing a wide variety of materials, femtosecond lasers are now used routinely. Understanding the interactions between the intense femtosecond pulse and the material could lead to technologically important advances. Pulse shaping could then be used to optimize the desired outcome. The scope of our research program is to develop robust and efficient strategies to control nonlinear laser-matter interactions using ultrashort shaped pulses in gas and condensed phases. Our systematic research has led to significant developments in a number of areas relevant to the AMO Physics group at DOE, among them: generation of ultrashort phase shaped pulses, coherent control and manipulation of quantum mechanical states in gas and condensed phases, behavior of isolated molecules under intense laser fields, behavior of condensed phase matter under intense laser field and implications on micromachining with ultrashort pulses, coherent control of nanoparticles their surface plasmon waves and their nonlinear optical behavior, and observation of coherent Coulomb explosion processes at 10 16 W/cm 2 . In all, the research has resulted in 36 publications (five journal covers) and nine invention disclosures, five of which have continued on to patenting

  12. Considerable improvement of entanglement swapping by considering multiphoton transitions via cavity quantum electrodynamics method

    Science.gov (United States)

    Pakniat, R.; Soltani, M.; Tavassoly, M. K.

    2018-03-01

    Recently we studied the effect of photon addition in the initial coherent field on the entanglement swapping which causes some improvements in the process [Soltani et al., Int. J. Mod. Phys. B 31, 1750198 (2017)]. In this paper, we investigate the influence of multiphoton transitions in the atom-field interaction based on the cavity quantum electrodynamics on the entanglement swapping and show its considerable constructive effect on this process. The presented model consists of two two-level atoms namely A1 and A2 and two distinct cavity fields F1 and F2. Initially, the atoms are prepared in a maximally entangled state and the fields in the cavities are prepared in hybrid entangled state of number and coherent states, separately. Making the atom A2 to interact with the field F1 (via the generalized Jaynes-Cummings model which allows m-photon transitions between atomic levels in the emission and absorption processes) followed by their detection allows us to arrive at the entanglement swapping from the two atoms A1, A2 and the two fields F1, F2 to the atom-field A1-F2 system. Then, we pay our attention to the time evolution of success probability of detecting processes and fidelity. Also, to determine the amount of entanglement of the generated entangled state in the swapping process, the linear entropy is evaluated and the effect of parameter m concerning the multiphoton transitions on these quantities is investigated, numerically. It is observed that, by increasing the number of photons in the transition process, one may obtain considerable improvement in the relevant quantities of the entanglement swapping. In detail, the satisfactorily acceptable values 1 and 0.5 corresponding to success probability and fidelity are obtained for most of the times during observing of the above-mentioned procedure. We concluded that the presented formalism in this paper is much more advantageous than our presentation model in our earlier work mentioned above.

  13. ACTS 2014

    DEFF Research Database (Denmark)

    Co-curator of ACTS 2014 together with Rasmus Holmboe, Judith Schwarzbart and Sanne Kofoed. ACTS is the Museum of Contemporary Art’s international bi-annual festival. ACTS was established in 2011 and, while the primary focus is on sound and performance art, it also looks toward socially oriented art....... For the 2014 festival, the museum has entered into a collaboration with the Department for Performance Design at Roskilde University – with continued focus on sound and performance art, and social art in public spaces. With ACTS, art moves out of its usual exhibition space and instead utilizes the city, its...... various possibilities and public spaces as a stage. ACTS takes place in and around the museum and diverse locations in Roskilde city. ACTS is partly curated by the museum staff and partly by guest curators. ACTS 2014 is supported by Nordea-fonden and is a part of the project The Museum goes downtown....

  14. The use of spinning-disk confocal microscopy for the intravital analysis of platelet dynamics in response to systemic and local inflammation.

    Directory of Open Access Journals (Sweden)

    Craig N Jenne

    Full Text Available Platelets are central players in inflammation and are an important component of the innate immune response. The ability to visualize platelets within the live host is essential to understanding their role in these processes. Past approaches have involved adoptive transfer of labelled platelets, non-specific dyes, or the use of fluorescent antibodies to tag platelets in vivo. Often, these techniques result in either the activation of the platelet, or blockade of specific platelet receptors. In this report, we describe two new methods for intravital visualization of platelet biology, intravenous administration of labelled anti-CD49b, which labels all platelets, and CD41-YFP transgenic mice, in which a percentage of platelets express YFP. Both approaches label endogenous platelets and allow for their visualization using spinning-disk confocal fluorescent microscopy. Following LPS-induced inflammation, we were able to measure a significant increase in both the number and size of platelet aggregates observed within the vasculature of a number of different tissues. Real-time observation of these platelet aggregates reveals them to be large, dynamic structures that are continually expanding and sloughing-off into circulation. Using these techniques, we describe for the first time, platelet recruitment to, and behaviour within numerous tissues of the mouse, both under control conditions and following LPS induced inflammation.

  15. Extending the fundamental imaging-depth limit of multi-photon microscopy by imaging with photo-activatable fluorophores.

    Science.gov (United States)

    Chen, Zhixing; Wei, Lu; Zhu, Xinxin; Min, Wei

    2012-08-13

    It is highly desirable to be able to optically probe biological activities deep inside live organisms. By employing a spatially confined excitation via a nonlinear transition, multiphoton fluorescence microscopy has become indispensable for imaging scattering samples. However, as the incident laser power drops exponentially with imaging depth due to scattering loss, the out-of-focus fluorescence eventually overwhelms the in-focal signal. The resulting loss of imaging contrast defines a fundamental imaging-depth limit, which cannot be overcome by increasing excitation intensity. Herein we propose to significantly extend this depth limit by multiphoton activation and imaging (MPAI) of photo-activatable fluorophores. The imaging contrast is drastically improved due to the created disparity of bright-dark quantum states in space. We demonstrate this new principle by both analytical theory and experiments on tissue phantoms labeled with synthetic caged fluorescein dye or genetically encodable photoactivatable GFP.

  16. Imaging the morphological change of tissue structure during the early phase of esophageal tumor progression using multiphoton microscopy

    Science.gov (United States)

    Xu, Jian; Kang, Deyong; Xu, Meifang; Zhu, Xiaoqin; Zhuo, Shuangmu; Chen, Jianxin

    2012-12-01

    Esophageal cancer is a common malignancy with a very poor prognosis. Successful strategies for primary prevention and early detection are critically needed to control this disease. Multiphoton microscopy (MPM) is becoming a novel optical tool of choice for imaging tissue architecture and cellular morphology by two-photon excited fluorescence. In this study, we used MPM to image microstructure of human normal esophagus, carcinoma in situ (CIS), and early invasive carcinoma in order to establish the morphological features to differentiate these tissues. The diagnostic features such as the appearance of cancerous cells, the significant loss of stroma, the absence of the basement membrane were extracted to distinguish between normal and cancerous esophagus tissue. These results correlated well with the paired histological findings. With the advancement of clinically miniaturized MPM and the multi-photon probe, combining MPM with standard endoscopy will therefore allow us to make a real-time in vivo diagnosis of early esophageal cancer at the cellular level.

  17. Controlling the transmitted information of a multi-photon interacting with a single-Cooper pair box

    Energy Technology Data Exchange (ETDEWEB)

    Kadry, Heba, E-mail: hkadry1@yahoo.com; Abdel-Aty, Abdel-Haleem, E-mail: hkadry1@yahoo.com; Zakaria, Nordin, E-mail: hkadry1@yahoo.com [Computer and Information Science Department, Universiti Teknologi Petronas, Seri Iskandar, 31750 Tronoh, Perak (Malaysia); Cheong, Lee Yen [Fundamental and Applied Science Department, Universiti Teknologi Petronas, Seri Iskandar, 31750 Tronoh, Perak (Malaysia)

    2014-10-24

    We study a model of a multi-photon interaction of a single Cooper pair box with a cavity field. The exchange of the information using this system is studied. We quantify the fidelity of the transmitted information. The effect of the system parameters (detuning parameter, field photons, state density and mean photon number) in the fidelity of the transmitted information is investigated. We found that the fidelity of the transmitted information can be controlled using the system parameters.

  18. Controlling the transmitted information of a multi-photon interacting with a single-Cooper pair box

    International Nuclear Information System (INIS)

    Kadry, Heba; Abdel-Aty, Abdel-Haleem; Zakaria, Nordin; Cheong, Lee Yen

    2014-01-01

    We study a model of a multi-photon interaction of a single Cooper pair box with a cavity field. The exchange of the information using this system is studied. We quantify the fidelity of the transmitted information. The effect of the system parameters (detuning parameter, field photons, state density and mean photon number) in the fidelity of the transmitted information is investigated. We found that the fidelity of the transmitted information can be controlled using the system parameters

  19. Visible luminescence studies in the infrared multiphoton dissociation of 1,2-dichloro-1,1-difluoroethane

    Science.gov (United States)

    Pushpa, K. K.; Kumar, Awadesh; Naik, P. D.; Annaji Rao, K.; Parthasarathy, V.; Sarkar, S. K.; Mittal, J. P.

    1997-11-01

    A strong visible luminescence was observed in the CO 2 laser induced infrared multiphoton dissociation of 1,2-dichloro-1,1-difluoroethane. The emission observed between 350-750 nm is attributed to electronically excited carbene CF 2ClCH. The temporal profile of this luminescence was studied as a function of laser pulse duration, pulse energy, excitation frequency and substrate pressure. A suitable dissociation mechanism is presented considering various channels of IRMPD of this molecule.

  20. S-matrix analysis of vibrational and alignment effects in intense-field multiphoton ionization of molecules

    Energy Technology Data Exchange (ETDEWEB)

    Requate, A

    2007-03-15

    Theoretical analysis of the vibrational excitation of small molecules during multiphoton ionization in intense laser fields of optical and infrared frequencies. Analysis of the alignment dependence of the electron impact ionization of diatomic molecules in the presence of an intense laser field as the final step in the process of Nonsequential Double Ionization. Quantum mechanical description using S-matrix theory in Strong Field Approximation (SFA), i.e. beyond perturbation theory. (orig.)

  1. S-matrix analysis of vibrational and alignment effects in intense-field multiphoton ionization of molecules

    International Nuclear Information System (INIS)

    Requate, A.

    2007-03-01

    Theoretical analysis of the vibrational excitation of small molecules during multiphoton ionization in intense laser fields of optical and infrared frequencies. Analysis of the alignment dependence of the electron impact ionization of diatomic molecules in the presence of an intense laser field as the final step in the process of Nonsequential Double Ionization. Quantum mechanical description using S-matrix theory in Strong Field Approximation (SFA), i.e. beyond perturbation theory. (orig.)

  2. Curatorial Acts

    NARCIS (Netherlands)

    Bal, M.

    2012-01-01

    In a self-critical inquiry into my own recent work of co-curating and the experience of seeing my video work being curated by others, this article examines acts of framing as performative acts that seek to transform visitors' preconceptions. This affective effect is pursued by means of immersion,

  3. Multiphoton minimal inertia scanning for fast acquisition of neural activity signals

    Science.gov (United States)

    Schuck, Renaud; Go, Mary Ann; Garasto, Stefania; Reynolds, Stephanie; Dragotti, Pier Luigi; Schultz, Simon R.

    2018-04-01

    Objective. Multi-photon laser scanning microscopy provides a powerful tool for monitoring the spatiotemporal dynamics of neural circuit activity. It is, however, intrinsically a point scanning technique. Standard raster scanning enables imaging at subcellular resolution; however, acquisition rates are limited by the size of the field of view to be scanned. Recently developed scanning strategies such as travelling salesman scanning (TSS) have been developed to maximize cellular sampling rate by scanning only select regions in the field of view corresponding to locations of interest such as somata. However, such strategies are not optimized for the mechanical properties of galvanometric scanners. We thus aimed to develop a new scanning algorithm which produces minimal inertia trajectories, and compare its performance with existing scanning algorithms. Approach. We describe here the adaptive spiral scanning (SSA) algorithm, which fits a set of near-circular trajectories to the cellular distribution to avoid inertial drifts of galvanometer position. We compare its performance to raster scanning and TSS in terms of cellular sampling frequency and signal-to-noise ratio (SNR). Main Results. Using surrogate neuron spatial position data, we show that SSA acquisition rates are an order of magnitude higher than those for raster scanning and generally exceed those achieved by TSS for neural densities comparable with those found in the cortex. We show that this result also holds true for in vitro hippocampal mouse brain slices bath loaded with the synthetic calcium dye Cal-520 AM. The ability of TSS to ‘park’ the laser on each neuron along the scanning trajectory, however, enables higher SNR than SSA when all targets are precisely scanned. Raster scanning has the highest SNR but at a substantial cost in number of cells scanned. To understand the impact of sampling rate and SNR on functional calcium imaging, we used the Cramér-Rao Bound on evoked calcium traces recorded

  4. Multimodal microscopy and the stepwise multi-photon activation fluorescence of melanin

    Science.gov (United States)

    Lai, Zhenhua

    The author's work is divided into three aspects: multimodal microscopy, stepwise multi-photon activation fluorescence (SMPAF) of melanin, and customized-profile lenses (CPL) for on-axis laser scanners, which will be introduced respectively. A multimodal microscope provides the ability to image samples with multiple modalities on the same stage, which incorporates the benefits of all modalities. The multimodal microscopes developed in this dissertation are the Keck 3D fusion multimodal microscope 2.0 (3DFM 2.0), upgraded from the old 3DFM with improved performance and flexibility, and the multimodal microscope for targeting small particles (the "Target" system). The control systems developed for both microscopes are low-cost and easy-to-build, with all components off-the-shelf. The control system have not only significantly decreased the complexity and size of the microscope, but also increased the pixel resolution and flexibility. The SMPAF of melanin, activated by a continuous-wave (CW) mode near-infrared (NIR) laser, has potential applications for a low-cost and reliable method of detecting melanin. The photophysics of melanin SMPAF has been studied by theoretical analysis of the excitation process and investigation of the spectra, activation threshold, and photon number absorption of melanin SMPAF. SMPAF images of melanin in mouse hair and skin, mouse melanoma, and human black and white hairs are compared with images taken by conventional multi-photon fluorescence microscopy (MPFM) and confocal reflectance microscopy (CRM). SMPAF images significantly increase specificity and demonstrate the potential to increase sensitivity for melanin detection compared to MPFM images and CRM images. Employing melanin SMPAF imaging to detect melanin inside human skin in vivo has been demonstrated, which proves the effectiveness of melanin detection using SMPAF for medical purposes. Selective melanin ablation with micrometer resolution has been presented using the Target system

  5. Graphene oxide from silk cocoon: a novel magnetic fluorophore for multi-photon imaging.

    Science.gov (United States)

    Roy, Manas; Kusurkar, Tejas Sanjeev; Maurya, Sandeep Kumar; Meena, Sunil Kumar; Singh, Sushil Kumar; Sethy, Niroj; Bhargava, Kalpana; Sharma, Raj Kishore; Goswami, Debabrata; Sarkar, Sabyasachi; Das, Mainak

    2014-02-01

    In this work, we synthesized graphene oxide from silk cocoon embarking its new dimension as a magnetic fluorophore when compared with its present technical status, which at best is for extracting silk as a biomaterial for tissue engineering applications. We produced graphene oxide by pyrolysing the silk cocoon in an inert atmosphere. The collected raw carbon is oxidized by nitric acid that readily produces multilayer graphene oxide with nano carbon particulates. Structural properties of the graphene oxide were analyzed using scanning electron microscopy, transmission electron microscopy, Fourier transform infra-red spectroscopy, and Raman spectroscopy. The oxidized sample shows remarkable fluorescence, multi-photon imaging and magnetic properties. On increasing the excitation wavelength, the fluorescence emission intensity of the graphene oxide also increases and found maximum emission at 380 nm excitation wavelength. On studying the two photon absorption (TPA) property of aqueous graphene oxide using Z-scan technique, we found significant TPA activity at near infrared wavelength. In addition, the graphene oxide shows ferromagnetic behavior at room temperature. The observed fluorescence and magnetic property were attributed to the defects caused in the graphene oxide structure by introducing oxygen containing hydrophilic groups during the oxidation process. Previously silk cocoon has been used extensively in deriving silk-based tissue engineering materials and as gas filter. Here we show a novel application of silk cocoon by synthesizing graphene oxide based magnetic-fluorophore for bio-imaging applications.

  6. Non-descanned multifocal multiphoton microscopy with a multianode photomultiplier tube

    International Nuclear Information System (INIS)

    Cha, Jae Won; Yew, Elijah Y. S.; Kim, Daekeun; Subramanian, Jaichandar; Nedivi, Elly; So, Peter T. C.

    2015-01-01

    Multifocal multiphoton microscopy (MMM) improves imaging speed over a point scanning approach by parallelizing the excitation process. Early versions of MMM relied on imaging detectors to record emission signals from multiple foci simultaneously. For many turbid biological specimens, the scattering of emission photons results in blurred images and degrades the signal-to-noise ratio (SNR). We have recently demonstrated that a multianode photomultiplier tube (MAPMT) placed in a descanned configuration can effectively collect scattered emission photons from each focus into their corresponding anodes significantly improving image SNR for highly scattering specimens. Unfortunately, a descanned MMM has a longer detection path resulting in substantial emission photon loss. Optical design constraints in a descanned geometry further results in significant optical aberrations especially for large field-of-view (FOV), high NA objectives. Here, we introduce a non-descanned MMM based on MAPMT that substantially overcomes most of these drawbacks. We show that we improve signal efficiency up to fourfold with limited image SNR degradation due to scattered emission photons. The excitation foci can also be spaced wider to cover the full FOV of the objective with minimal aberrations. The performance of this system is demonstrated by imaging interneuron morphological structures deep in the brains of living mice

  7. Non-descanned multifocal multiphoton microscopy with a multianode photomultiplier tube

    Energy Technology Data Exchange (ETDEWEB)

    Cha, Jae Won; Yew, Elijah Y. S. [Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA (United States); Kim, Daekeun [Department of Mechanical Engineering, Dankook University (Korea, Republic of); Subramanian, Jaichandar [Picower Institute for Learning and Memory, Massachusetts Institute of Technology, Cambridge, MA (United States); Nedivi, Elly [Picower Institute for Learning and Memory, Massachusetts Institute of Technology, Cambridge, MA (United States); Departments of Brain and Cognitive Sciences, and Biology, Massachusetts Institute of Technology, Cambridge, MA (United States); So, Peter T. C. [Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA (United States); Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA (United States)

    2015-08-15

    Multifocal multiphoton microscopy (MMM) improves imaging speed over a point scanning approach by parallelizing the excitation process. Early versions of MMM relied on imaging detectors to record emission signals from multiple foci simultaneously. For many turbid biological specimens, the scattering of emission photons results in blurred images and degrades the signal-to-noise ratio (SNR). We have recently demonstrated that a multianode photomultiplier tube (MAPMT) placed in a descanned configuration can effectively collect scattered emission photons from each focus into their corresponding anodes significantly improving image SNR for highly scattering specimens. Unfortunately, a descanned MMM has a longer detection path resulting in substantial emission photon loss. Optical design constraints in a descanned geometry further results in significant optical aberrations especially for large field-of-view (FOV), high NA objectives. Here, we introduce a non-descanned MMM based on MAPMT that substantially overcomes most of these drawbacks. We show that we improve signal efficiency up to fourfold with limited image SNR degradation due to scattered emission photons. The excitation foci can also be spaced wider to cover the full FOV of the objective with minimal aberrations. The performance of this system is demonstrated by imaging interneuron morphological structures deep in the brains of living mice.

  8. Oxygen isotope separation by isotopically selective infrared multiphoton dissociation of 2,3-dihydropyran

    International Nuclear Information System (INIS)

    Yokoyama, Atsushi; Ohba, Hironori; Akagi, Hiroshi; Yokoyama, Keiichi; Saeki, Morihisa; Katsumata, Keiichi

    2008-01-01

    Oxygen isotopic selectivity on infrared multiphoton dissociation of 2,3-dihydropyran has been studied by the examination of the effects of excitation frequency, laser fluence, and gas pressure on the dissociation probability of 2,3-dihydropyran and isotopic composition of products. Oxygen-18 was enriched in a dissociation product: 2-propenal. The enrichment factor of 18 O and the dissociation probability were measured at laser frequency between 1033.5 and 1057.3 cm -1 ; the laser fluence of 2.2 - 2.3 J/cm 2 ; and the 2,3-dihydropyran pressure of 0.27 kPa. The dissociation probability decreases as the laser frequency being detuned from the absorption peak of 2,3-dihydropyran around 1081 cm -1 . On the other hand, the enrichment factor increases with detuning the frequency. The enrichment factor of 18 O increases with increasing the 2,3-dihydropyran pressure at the laser fluence below 3 J/cm 2 and the laser frequency of 1033.5 cm -1 , whereas the yield of 2-propenal decreases with increasing the pressure. Very high enrichment factor of 751 was obtained by the irradiation of 0.53 kPa of 2,3-dihydropyran at 2.1 J/cm 2 . (author)

  9. Population inversion of two atoms under the phase decoherence in the multiphoton process

    International Nuclear Information System (INIS)

    Zhang Dongxia; Sa Chuerfu; Mu Qier

    2011-01-01

    By means of the quantum theory, the population inversion of two atoms in the system of two two-level atoms coupled to a light field in the Binomial Optical Field are investigated in the presence of phase decoherence in the multiphoton Tavis-Cumming Model. The influences of the phase decoherence coefficient, the parameters η of the binomial optical field, the maximum number of photons and the number of the transitional photons on the properties of the population inversion of two atoms have been discussed. The results show that the phase decoherence reduced the oscillation amplitude of the population inversion of two atoms and destroyed the atomic quantum characteristic. Changing the number of the transitional photons, evolved cycle and evolved intensity the population inversion of two atoms can be changed. The phenomena of collapse and revival disappear as photon number increase. When the binomial optical state changes from a coherent state to a Fock state, the oscillation frequency of the atomic population reduces gradually, the phenomena of collapse and revival vanishes gradually. (authors)

  10. Proximal design for a multimodality endoscope with multiphoton microscopy, optical coherence microscopy and visual modalities

    Science.gov (United States)

    Kiekens, Kelli C.; Talarico, Olivia; Barton, Jennifer K.

    2018-02-01

    A multimodality endoscope system has been designed for early detection of ovarian cancer. Multiple illumination and detection systems must be integrated in a compact, stable, transportable configuration to meet the requirements of a clinical setting. The proximal configuration presented here supports visible light navigation with a large field of view and low resolution, high resolution multiphoton microscopy (MPM), and high resolution optical coherence microscopy (OCM). All modalities are integrated into a single optical system in the endoscope. The system requires two light sources: a green laser for visible light navigation and a compact fiber based femtosecond laser for MPM and OCM. Using an inline wavelength division multiplexer, the two sources are combined into a single mode fiber. To accomplish OCM, a fiber coupler is used to separate the femtosecond laser into a reference arm and signal arm. The reflected reference arm and the signal from the sample are interfered and wavelength separated by a reflection grating and detected using a linear array. The MPM signal is collimated and goes through a series of filters to separate the 2nd and 3rd harmonics as well as twophoton excitation florescence (2PEF) and 3PEF. Each signal is independently detected on a photo multiplier tube and amplified. The visible light is collected by multiple high numerical aperture fibers at the endoscope tip which are bundled into one SMA adapter at the proximal end and connected to a photodetector. This integrated system design is compact, efficient and meets both optical and mechanical requirements for clinical applications.

  11. Quantification of aortic and cutaneous elastin and collagen morphology in Marfan syndrome by multiphoton microscopy.

    Science.gov (United States)

    Cui, Jason Z; Tehrani, Arash Y; Jett, Kimberly A; Bernatchez, Pascal; van Breemen, Cornelis; Esfandiarei, Mitra

    2014-09-01

    In a mouse model of Marfan syndrome, conventional Verhoeff-Van Gieson staining displays severe fragmentation, disorganization and loss of the aortic elastic fiber integrity. However, this method involves chemical fixatives and staining, which may alter the native morphology of elastin and collagen. Thus far, quantitative analysis of fiber damage in aorta and skin in Marfan syndrome has not yet been explored. In this study, we have used an advanced noninvasive and label-free imaging technique, multiphoton microscopy to quantify fiber fragmentation, disorganization, and total volumetric density of aortic and cutaneous elastin and collagen in a mouse model of Marfan syndrome. Aorta and skin samples were harvested from Marfan and control mice aged 3-, 6- and 9-month. Elastin and collagen were identified based on two-photon excitation fluorescence and second-harmonic-generation signals, respectively, without exogenous label. Measurement of fiber length indicated significant fragmentation in Marfan vs. control. Fast Fourier transform algorithm analysis demonstrated markedly lower fiber organization in Marfan mice. Significantly reduced volumetric density of elastin and collagen and thinner skin dermis were observed in Marfan mice. Cutaneous content of elastic fibers and thickness of dermis in 3-month Marfan resembled those in the oldest control mice. Our findings of early signs of fiber degradation and thinning of skin dermis support the potential development of a novel non-invasive approach for early diagnosis of Marfan syndrome. Copyright © 2014 Elsevier Inc. All rights reserved.

  12. Evaluation of endogenous species involved in brain tumors using multiphoton photoacoustic spectroscopy

    Science.gov (United States)

    Dahal, Sudhir; Cullum, Brian M.

    2013-05-01

    It has been shown that using non-resonant multiphoton photoacoustic spectroscopy (NMPPAS), excised brain tumor (grade III astrocytoma) and healthy tissue can be differentiated from each other, even in neighboring biopsy samples[1, 2]. Because of this, this powerful technique offers a great deal of potential for use as a surgical guidance technique for tumor margining with up to cellular level spatial resolution[3]. NMPPAS spectra are obtained by monitoring the non-radiative relaxation pathways via ultrasonic detection, following two-photon excitation with light in the optical diagnostic window (740nm-1100nm). Based upon significant differences in the ratiometric absorption of the tissues following 970nm and 1100nm excitation, a clear classification of the tissue can be made. These differences are the result of variations in composition and oxidation state of certain endogenous biochemical species between healthy and malignant tissues. In this work, NADH, NAD+ and ATP were measured using NMPPAS in model gelatin tissue phantoms to begin to understand which species might be responsible for the observed spectral differences in the tissue. Each species was placed in specific pH environments to provide control over the ratio of oxidized to reduced forms of the species. Ratiometric analyses were then conducted to account for variability caused due to instrumental parameters. This paper will discuss the potential roles of each of the species for tumor determination and their contribution to the spectral signature.

  13. Multiphoton ionization of (Xe)n and (NO)n clusters using a picosecond laser

    International Nuclear Information System (INIS)

    Smith, D.B.; Miller, J.C.

    1989-01-01

    Mass-resolved multiphoton ionization (MPI) spectroscopy is an established technique for detecting and analyzing van der Waals molecules and larger clusters. MPI spectroscopy provides excellent detection sensitivity, moderately high resolution, and selectivity among cluster species. In addition to information provided by the analysis of photoions following MPI, photoelectron spectroscopy can reveal details regarding the structure of ionic states. Unfortunately, the technique is limited by its tendency to produce extensive fragmentation. Fragmentation is also a problem with other ionization techniques (e.g., electron impact ionization), but the intense laser beams required for MPI cause additional dissociation channels to become available. These channels include absorption of additional photons by parent ions (ion ladder mechanism), absorption of additional photons by fragment ions (ladder switching mechanism), and resonances with dissociative states in the neutral manifold. The existence of these dissociation channels can preclude the use of MPI spectroscopy in many situations. Recently, MPI studies of stable molecules using picosecond lasers (pulse length = 1 - 10 ps) have indicated that limitations due to fragmentation might be subdued. With picosecond lasers, dissociation mechanisms can be altered and in some cases fragmentation can be eliminated or reduced. Additional photon absorption competes effectively with dissociation channels when a very short laser pulse or, perhaps more importantly, a sufficiently high peak-power is used. In the case where ionic absorption and fragmentation occurs, it has been shown that picosecond MPI might favor the ion ladder mechanism rather than the ladder switching mechanism

  14. Identifying Two Common Types of Breast Benign Diseases Based on Multiphoton Microscopy

    Directory of Open Access Journals (Sweden)

    Yan Wu

    2018-01-01

    Full Text Available Multiphoton microscopy has attracted increasing attention and investigations in the field of breast cancer, based on two-photon excited fluorescence (TPEF and second-harmonic generation (SHG. However, the incidence of breast benign diseases is about 5 to 10 times higher than breast cancer; up to 30% of women suffer from breast benign diseases and require treatment at some time in their lives. Thus, in this study, MPM was applied to image fibroadenoma and fibrocystic lesion, which are two of the most common breast benign diseases. The results show that MPM has the capability to identify the microstructure of lobule and stroma in normal breast tissue, the interaction of compressed ducts with surrounding collagen fiber in fibroadenoma, and the architecture of cysts filled with cystic fluid in fibrocystic disease. These findings indicate that, with integration of MPM into currently accepted clinical imaging system, it has the potential to make a real-time diagnosis of breast benign diseases in vivo, as well as breast cancer.

  15. Multiphoton microscopy as a diagnostic imaging modality for pancreatic neoplasms without hematoxylin and eosin stains

    Science.gov (United States)

    Chen, Youting; Chen, Jing; Chen, Hong; Hong, Zhipeng; Zhu, Xiaoqin; Zhuo, Shuangmu; Chen, Yanling; Chen, Jianxin

    2014-09-01

    Hematoxylin and eosin (H&E) staining of tissue samples is the standard approach in histopathology for imaging and diagnosing cancer. Recent reports have shown that multiphoton microscopy (MPM) provides better sample interface with single-cell resolution, which enhances traditional H&E staining and offers a powerful diagnostic tool with potential applications in oncology. The purpose of this study was to further expand the versatility of MPM by establishing the optical parameters required for imaging unstained histological sections of pancreatic neoplasms, thereby providing an efficient and environmentally sustainable alternative to H&E staining while improving the accuracy of pancreatic cancer diagnoses. We found that the high-resolution MPM images clearly distinguish between the structure of normal pancreatic tissues compared with pancreatic neoplasms in unstained histological sections, and discernable differences in tissue architecture and cell morphology between normal versus tumorigenic cells led to enhanced optical diagnosis of cancerous tissue. Moreover, quantitative assessment of the cytomorphological features visualized from MPM images showed significant differences in the nuclear-cytoplasmic ratios of pancreatic neoplasms compared with normal pancreas, as well as further distinguished pancreatic malignant tumors from benign tumors. These results indicate that the MPM could potentially serve as an optical tool for the diagnosis of pancreatic neoplasms in unstained histological sections.

  16. Full optical model of micro-endoscope with optical coherence microscopy, multiphoton microscopy and visible capabilities

    Science.gov (United States)

    Vega, David; Kiekens, Kelli C.; Syson, Nikolas C.; Romano, Gabriella; Baker, Tressa; Barton, Jennifer K.

    2018-02-01

    While Optical Coherence Microscopy (OCM), Multiphoton Microscopy (MPM), and narrowband imaging are powerful imaging techniques that can be used to detect cancer, each imaging technique has limitations when used by itself. Combining them into an endoscope to work in synergy can help achieve high sensitivity and specificity for diagnosis at the point of care. Such complex endoscopes have an elevated risk of failure, and performing proper modelling ensures functionality and minimizes risk. We present full 2D and 3D models of a multimodality optical micro-endoscope to provide real-time detection of carcinomas, called a salpingoscope. The models evaluate the endoscope illumination and light collection capabilities of various modalities. The design features two optical paths with different numerical apertures (NA) through a single lens system with a scanning optical fiber. The dual path is achieved using dichroic coatings embedded in a triplet. A high NA optical path is designed to perform OCM and MPM while a low NA optical path is designed for the visible spectrum to navigate the endoscope to areas of interest and narrowband imaging. Different tests such as the reflectance profile of homogeneous epithelial tissue were performed to adjust the models properly. Light collection models for the different modalities were created and tested for efficiency. While it is challenging to evaluate the efficiency of multimodality endoscopes, the models ensure that the system is design for the expected light collection levels to provide detectable signal to work for the intended imaging.

  17. Open-ended response theory with polarizable embedding: multiphoton absorption in biomolecular systems.

    Science.gov (United States)

    Steindal, Arnfinn Hykkerud; Beerepoot, Maarten T P; Ringholm, Magnus; List, Nanna Holmgaard; Ruud, Kenneth; Kongsted, Jacob; Olsen, Jógvan Magnus Haugaard

    2016-10-12

    We present the theory and implementation of an open-ended framework for electric response properties at the level of Hartree-Fock and Kohn-Sham density functional theory that includes effects from the molecular environment modeled by the polarizable embedding (PE) model. With this new state-of-the-art multiscale functionality, electric response properties to any order can be calculated for molecules embedded in polarizable atomistic molecular environments ranging from solvents to complex heterogeneous macromolecules such as proteins. In addition, environmental effects on multiphoton absorption (MPA) properties can be studied by evaluating single residues of the response functions. The PE approach includes mutual polarization effects between the quantum and classical parts of the system through induced dipoles that are determined self-consistently with respect to the electronic density. The applicability of our approach is demonstrated by calculating MPA strengths up to four-photon absorption for the green fluorescent protein. We show how the size of the quantum region, as well as the treatment of the border between the quantum and classical regions, is crucial in order to obtain reliable MPA predictions.

  18. Gas chromatography/multiphoton ionization/time-of-flight mass spectrometry of polychlorinated biphenyls

    International Nuclear Information System (INIS)

    Matsui, Taiki; Uchimura, Tomohiro; Imasaka, Totaro

    2011-01-01

    A sample mixture of polychlorinated biphenyls (PCBs) was measured by gas chromatography/multiphoton ionization/time-of-flight mass spectrometry (GC/MPI/TOF-MS) using four types of laser sources. When a fourth harmonic emission (266 nm) of a picosecond Nd:YAG laser (1064 nm) was utilized, highly chlorinated PCBs larger than hepta-CBs were not observed. A fifth harmonic emission (213 nm) of the picosecond Nd:YAG laser allowed the measurement of PCBs from di-CBs to octa-CBs, and the limit of detection (LOD) was several pg for each component of PCBs. The LOD for the total amount of PCBs, which was calculated using the protocol provided by the Ministry of the Environment, Japan, was 1000 pg. The signal intensity of the congeners with chlorine atoms at the ortho positions (non-coplanar PCBs) was enhanced by using the fifth harmonic emission. When the fourth harmonic emission remaining after fifth harmonic generation was simultaneously used, the LOD for total PCBs was improved to 667 pg. The PCB sample was also measured using a third harmonic emission (267 nm) of a femtosecond Ti:sapphire laser (800 nm), providing an LOD of 677 pg. Thus, the two-color beam (266/213 nm) of a picosecond Nd:YAG laser had a comparable, or even slightly superior, performance to the more expensive femtosecond Ti:sapphire laser.

  19. Clinical studies of pigmented lesions in human skin by using a multiphoton tomograph

    Science.gov (United States)

    Balu, Mihaela; Kelly, Kristen M.; Zachary, Christopher B.; Harris, Ronald M.; Krasieva, Tatiana B.; König, Karsten; Tromberg, Bruce J.

    2013-02-01

    In vivo imaging of pigmented lesions in human skin was performed with a clinical multiphoton microscopy (MPM)-based tomograph (MPTflex, JenLab, Germany). Two-photon excited fluorescence was used for visualizing endogenous fluorophores such as NADH/FAD, keratin, melanin in the epidermal cells and elastin fibers in the dermis. Collagen fibers were imaged by second harmonic generation. Our study involved in vivo imaging of benign melanocytic nevi, atypical nevi and melanoma. The goal of this preliminary study was to identify in vivo the characteristic features and their frequency in pigmented lesions at different stages (benign, atypical and malignant) and to evaluate the ability of in vivo MPM to distinguish atypical nevi from melanoma. Comparison with histopathology was performed for the biopsied lesions. Benign melanocytic nevi were characterized by the presence of nevus cell nests at the epidermal-dermal junction. In atypical nevi, features such as lentiginous hyperplasia, acanthosis and architectural disorder were imaged. Cytological atypia was present in all the melanoma lesions imaged, showing the strongest correlation with malignancy. The MPM images demonstrated very good correlation with corresponding histological images, suggesting that MPM could be a promising tool for in vivo non-invasive pigmented lesion diagnosis, particularly distinguishing atypical nevi from melanoma.

  20. Multimodal imaging of vocal fold scarring in a rabbit model by multiphoton microscopy

    Science.gov (United States)

    Kazarine, Alexei; Bouhabel, Sarah; Douillette, Annie H.; Kost, Karen; Li-Jessen, Nicole Y. K.; Mongeau, Luc; Wiseman, Paul W.

    2017-02-01

    Vocal fold scarring as a result of injury or disease can lead to voice disorders which can significantly affect the quality of life. During the scarring process, the normally elastic tissue of the vocal fold lamina propria is replaced by a much stiffer collagen-based fibrotic tissue, which impacts the fold's ability to vibrate. Surgical removal of this tissue is often ineffective and can result in further scarring. Injectable biomaterials, a form of tissue engineering, have been proposed as a potential solution to reduce existing scars or prevent scarring altogether. In order to properly evaluate the effectiveness of these new materials, multiphoton microscopy emerges as an effective tool due to its intrinsic multiple label free contrast mechanisms that highlight extracellular matrix elements. In this study, we evaluate the spatial distribution of collagen and elastin fibers in a rabbit model using second harmonic generation (SHG), third harmonic generation (THG) and two photon autofluorescence (TPAF) applied to unlabeled tissue sections. In comparison to traditional methods that rely on histological staining or immunohistochemistry, SHG, THG and TPAF provide a more reliable detection of these native proteins. The evaluation of collagen levels allows us to follow the extent of scarring, while the presence of elastin fibers is thought to be indicative of the level of healing of the injured fold. Using these imaging modalities, we characterize the outcome of injectable biomaterial treatments in order to direct future treatments for tissue engineering.

  1. Approach to quantify human dermal skin aging using multiphoton laser scanning microscopy

    Science.gov (United States)

    Puschmann, Stefan; Rahn, Christian-Dennis; Wenck, Horst; Gallinat, Stefan; Fischer, Frank

    2012-03-01

    Extracellular skin structures in human skin are impaired during intrinsic and extrinsic aging. Assessment of these dermal changes is conducted by subjective clinical evaluation and histological and molecular analysis. We aimed to develop a new parameter for the noninvasive quantitative determination of dermal skin alterations utilizing the high-resolution three-dimensional multiphoton laser scanning microscopy (MPLSM) technique. To quantify structural differences between chronically sun-exposed and sun-protected human skin, the respective collagen-specific second harmonic generation and the elastin-specific autofluorescence signals were recorded in young and elderly volunteers using the MPLSM technique. After image processing, the elastin-to-collagen ratio (ELCOR) was calculated. Results show that the ELCOR parameter of volar forearm skin significantly increases with age. For elderly volunteers, the ELCOR value calculated for the chronically sun-exposed temple area is significantly augmented compared to the sun-protected upper arm area. Based on the MPLSM technology, we introduce the ELCOR parameter as a new means to quantify accurately age-associated alterations in the extracellular matrix.

  2. Multiphoton ionization in superintense, high-frequency laser fields. I. General developments

    International Nuclear Information System (INIS)

    Pont, M.

    1991-01-01

    This is the first of two papers studying multiphoton ionization (MPI) in superintense, high-frequency laser fields. They are based on a general iteration scheme in increasing powers of the inverse frequency. To lowest order in the frequency, i.e., the high-frequency limit, the atom was shown to be stable against decay by MPI, though distorted. To next order in the iteration, an expression for the MPI amplitude was obtained. In the present paper, we present general developments from this expression, valid for arbitrary polarization, binding potential, intensity, and initial state. First we analyze the symmetry of the angular distributions of photoelectrons determined by this expression for the MPI amplitude. This expression can explain the asymmetries in the angular distributions of photoelectrons occurring in the case of elliptic polarization that were recently reported in experiments. In the radiation regime where our theory applies these asymmetries are, however, weak. In certain instances our theory yields asymmetries in cases where lowest-order perturbation theory (LOPT) fails to predict them. We prove that at low intensities our expression for the MPI amplitude yields results in agreement with LOPT evaluated at high frequencies. An important part of this paper consists, however, of the derivation of an alternative form for the MPI amplitude of atomic hydrogen, which is substantially simpler, though somewhat less accurate. We study the consequences of this simplified expression for the case of linearly polarized fields in the following paper [Phys. Rev. A 44, xxxx (1991)

  3. Time-resolved photoion imaging spectroscopy: Determining energy distribution in multiphoton absorption experiments

    Science.gov (United States)

    Qian, D. B.; Shi, F. D.; Chen, L.; Martin, S.; Bernard, J.; Yang, J.; Zhang, S. F.; Chen, Z. Q.; Zhu, X. L.; Ma, X.

    2018-04-01

    We propose an approach to determine the excitation energy distribution due to multiphoton absorption in the case of excited systems following decays to produce different ion species. This approach is based on the measurement of the time-resolved photoion position spectrum by using velocity map imaging spectrometry and an unfocused laser beam with a low fluence and homogeneous profile. Such a measurement allows us to identify the species and the origin of each ion detected and to depict the energy distribution using a pure Poisson's equation involving only one variable which is proportional to the absolute photon absorption cross section. A cascade decay model is used to build direct connections between the energy distribution and the probability to detect each ionic species. Comparison between experiments and simulations permits the energy distribution and accordingly the absolute photon absorption cross section to be determined. This approach is illustrated using C60 as an example. It may therefore be extended to a wide variety of molecules and clusters having decay mechanisms similar to those of fullerene molecules.

  4. Identification of tumor cells infiltrating into connective tissue in esophageal cancer by multiphoton microscopy

    Science.gov (United States)

    Xu, Jian; Jiang, Liwei; Kang, Deyong; Wu, Xuejing; Xu, Meifang; Zhuo, Shuangmu; Zhu, Xiaoqin; Lin, Jiangbo; Chen, Jianxin

    2016-10-01

    Esophageal cancer is one of the most common malignancies of the gastrointestinal cancers and carries poorer prognosis than other gastrointestinal cancers. In general practice, the depth of tumor infiltration in esophageal wall is crucial to establishing appropriate treatment plan which is established by detecting the tumor infiltration depth. Connective tissue is one of the main structures that form the esophageal wall. So, identification of tumor cells infiltrating into connective tissue is helping for detecting the tumor infiltration depth. Our aim is to evaluate whether multiphoton microscopy (MPM) can be used to detect tumor cells infiltrating into connective tissue in the esophageal cancer. MPM is well-suited for real-time detecting morphologic and cellular changes in fresh tissues since many endogenous fluorophores of fresh tissues are excited through two-photon excited fluorescence (TPEF) and second harmonic generation (SHG). In this work, microstructure of tumor cells and connective tissue are first studied. Then, morphological changes of collagen fibers after the infiltration of tumor cells are shown. These results show that MPM has the ability to detect tumor cells infiltrating into connective tissue in the esophageal cancer. In the future, MPM may be a promising imaging technique for detecting tumor cells in esophageal cancer.

  5. A phasor approach analysis of multiphoton FLIM measurements of three-dimensional cell culture models

    Science.gov (United States)

    Lakner, P. H.; Möller, Y.; Olayioye, M. A.; Brucker, S. Y.; Schenke-Layland, K.; Monaghan, M. G.

    2016-03-01

    Fluorescence lifetime imaging microscopy (FLIM) is a useful approach to obtain information regarding the endogenous fluorophores present in biological samples. The concise evaluation of FLIM data requires the use of robust mathematical algorithms. In this study, we developed a user-friendly phasor approach for analyzing FLIM data and applied this method on three-dimensional (3D) Caco-2 models of polarized epithelial luminal cysts in a supporting extracellular matrix environment. These Caco-2 based models were treated with epidermal growth factor (EGF), to stimulate proliferation in order to determine if FLIM could detect such a change in cell behavior. Autofluorescence from nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) in luminal Caco-2 cysts was stimulated by 2-photon laser excitation. Using a phasor approach, the lifetimes of involved fluorophores and their contribution were calculated with fewer initial assumptions when compared to multiexponential decay fitting. The phasor approach simplified FLIM data analysis, making it an interesting tool for non-experts in numerical data analysis. We observed that an increased proliferation stimulated by EGF led to a significant shift in fluorescence lifetime and a significant alteration of the phasor data shape. Our data demonstrates that multiphoton FLIM analysis with the phasor approach is a suitable method for the non-invasive analysis of 3D in vitro cell culture models qualifying this method for monitoring basic cellular features and the effect of external factors.

  6. All-optical bidirectional neural interfacing using hybrid multiphoton holographic optogenetic stimulation.

    Science.gov (United States)

    Paluch-Siegler, Shir; Mayblum, Tom; Dana, Hod; Brosh, Inbar; Gefen, Inna; Shoham, Shy

    2015-07-01

    Our understanding of neural information processing could potentially be advanced by combining flexible three-dimensional (3-D) neuroimaging and stimulation. Recent developments in optogenetics suggest that neurophotonic approaches are in principle highly suited for noncontact stimulation of network activity patterns. In particular, two-photon holographic optical neural stimulation (2P-HONS) has emerged as a leading approach for multisite 3-D excitation, and combining it with temporal focusing (TF) further enables axially confined yet spatially extended light patterns. Here, we study key steps toward bidirectional cell-targeted 3-D interfacing by introducing and testing a hybrid new 2P-TF-HONS stimulation path for accurate parallel optogenetic excitation into a recently developed hybrid multiphoton 3-D imaging system. The system is shown to allow targeted all-optical probing of in vitro cortical networks expressing channelrhodopsin-2 using a regeneratively amplified femtosecond laser source tuned to 905 nm. These developments further advance a prospective new tool for studying and achieving distributed control over 3-D neuronal circuits both in vitro and in vivo.

  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. Rapid creation of distant entanglement by multi-photon resonant fluorescence

    Science.gov (United States)

    Cohen, Guy Z.; Sham, L. J.

    2014-03-01

    We study a simple, effective and robust method for entangling two separate stationary quantum dot spin qubits with high fidelity using multi-photon Gaussian state. The fluorescence signals from the two dots interfere at a beam splitter. The bosonic nature of photons leads, in analogy with the Hong-Ou-Mandel (HOM) effect, to selective pairing of photon holes (photon absences in the fluorescent signals). By the HOM effect, two photon holes with the same polarization end up at the same beam splitter output. As a result, two odd photon number detections at the outgoing beams, which must correspond to two photon holes with different polarizations, herald entanglement creation. The robustness of the Gaussian states is evidenced by the ability to compensate for photon absorption and noise by a moderate increase in the number of photons at the input. We calculate the entanglement generation rate in the ideal, non-ideal and near-ideal detector regimes and find substantial improvement over single-photon schemes in all three regimes. Fast and efficient spin-spin entanglement creation can form the basis for a scalable quantum dot quantum computing network. Our predictions can be tested using current experimental capabilities. This research was supported by the U.S. Army Research Office MURI award W911NF0910406, by NSF grant PHY-1104446 and by ARO (IARPA, W911NF-08-1-0487). The authors thank D. G. Steel for useful discussions.

  9. Temporal focusing-based widefield multiphoton microscopy with spatially modulated illumination for biotissue imaging.

    Science.gov (United States)

    Chang, Chia-Yuan; Lin, Cheng-Han; Lin, Chun-Yu; Sie, Yong-Da; Hu, Yvonne Yuling; Tsai, Sheng-Feng; Chen, Shean-Jen

    2018-01-01

    A developed temporal focusing-based multiphoton excitation microscope (TFMPEM) has a digital micromirror device (DMD) which is adopted not only as a blazed grating for light spatial dispersion but also for patterned illumination simultaneously. Herein, the TFMPEM has been extended to implement spatially modulated illumination at structured frequency and orientation to increase the beam coverage at the back-focal aperture of the objective lens. The axial excitation confinement (AEC) of TFMPEM can be condensed from 3.0 μm to 1.5 μm for a 50 % improvement. By using the TFMPEM with HiLo technique as two structured illuminations at the same spatial frequency but different orientation, reconstructed biotissue images according to the condensed AEC structured illumination are shown obviously superior in contrast and better scattering suppression. Picture: TPEF images of the eosin-stained mouse cerebellar cortex by conventional TFMPEM (left), and the TFMPEM with HiLo technique as 1.09 μm -1 spatially modulated illumination at 90° (center) and 0° (right) orientations. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Identifying and quantifying the stromal fibrosis in muscularis propria of colorectal carcinoma by multiphoton microscopy

    Science.gov (United States)

    Chen, Sijia; Yang, Yinghong; Jiang, Weizhong; Feng, Changyin; Chen, Zhifen; Zhuo, Shuangmu; Zhu, Xiaoqin; Guan, Guoxian; Chen, Jianxin

    2014-10-01

    The examination of stromal fibrosis within colorectal cancer is overlooked, not only because the routine pathological examinations seem to focus more on tumour staging and precise surgical margins, but also because of the lack of efficient diagnostic methods. Multiphoton microscopy (MPM) can be used to study the muscularis stroma of normal and colorectal carcinoma tissue at the molecular level. In this work, we attempt to show the feasibility of MPM for discerning the microstructure of the normal human rectal muscle layer and fibrosis colorectal carcinoma tissue practicably. Three types of muscularis propria stromal fibrosis beneath the colorectal cancer infiltration were first observed through the MPM imaging system by providing intercellular microstructural details in fresh, unstained tissue samples. Our approach also presents the capability of quantifying the extent of stromal fibrosis from both amount and orientation of collagen, which may further characterize the severity of fibrosis. By comparing with the pathology analysis, these results show that the MPM has potential advantages in becoming a histological tool for detecting the stromal fibrosis and collecting prognosis evidence, which may guide subsequent therapy procedures for patients into good prognosis.

  11. Exact perturbation theory of multiphoton processes at high intensities. [Schroedinger equation, perturbation theory, matrix

    Energy Technology Data Exchange (ETDEWEB)

    Faisal, F H.M. [Bielefeld Univ. (Germany, F.R.). Fakultaet fuer Physik

    1976-06-11

    In this work the perturbation theory for multiphoton processes at high intensities is investigated and it is described an analytical method of summing the perturbation series to extract the contribution from all terms that give rise to the absorption of N photons by an atomic system. The method is first applied to the solution of a simple model problem and the result is confirmed by direct integration of the model Schroedinger equation. The usual lowest (nonvanishing)-order perturbation-theoretical calculation is also carried out for this model to demonstrate explicitly that the full result correctly reproduces that of the lowest-order theory in the limit of low intensity. The method is then extended to the case of an atomic system with well-developed spectrum (e.g. H atom) and the N-photon T-matrix is derived in terms of a ''photon matrix'' asub(N), for which a three-term recurrence relation is established. Next, from the vantage point of the general result obtained here, A probe is made into the nature of several approximate nonperturbative solutions that have appeared in the literature in the past. It is shown here that their applicability is severely restricted by the requirement of the essential spectral degeneracy of the atomic system. Finally, appendix A outlines a prescription of computing the photon matrix asub(N), which (as in the usual lowest-order perturbation-theoretical calculation)requires a knowledge of the eigenfunctions and eigenvalues of the atomic Hamiltonian only.

  12. ACT Test

    Science.gov (United States)

    ... to clot, the higher the degree of clotting inhibition. During surgery, the ACT is kept above a ... What is ECLS? An Introduction to Extracorporeal Life Support. University of Michigan Health System [On-line information]. ...

  13. Searching for new features of intravitality of hanging based on macro- and microscopic evaluation of the proximal attachment of the sternocleidomastoid muscle and the mastoid process of the temporal bone.

    Science.gov (United States)

    Szleszkowski, Ł; Hałoń, A; Thannhäuser, A; Jurek, T

    2015-01-01

    Assessment of the usefulness of intravital lesions in the proximal attachment of the sternocleidomastoid muscle and the mastoid process of the temporal bone in medico-legal evaluation of death by hanging. The study material was obtained from the bodies of 35 people who died by hanging. The control group comprised specimens collected from 30 people who died of non-traumatic causes. The structures under study were examined macro- and microscopically. The basic change which could be recognized as a marker of intravitality of hanging was the presence of a macroscopically extensive blotchy area of abundant ecchymosis in the proximal muscle attachment, similar to that found in the distal attachment, and the presence of abundant diffuse intraosseous ecchymoses in the mastoid process. None of the cases revealed any ecchymoses in the proximal attachment of the muscle that would be similar to those present in the distal attachment. Discolourations within the mastoid processes, macroscopically suggestive of extensive intraosseous effusions arising from the mechanism of stretching, were not confirmed by microscopic evaluation and occurred at the same frequency as in the control group. Limitations of the study were related to the method which involved sample collection by means of bone chisels, decalcification and preparation of specimens, which had an effect, for example, on the measurable evaluation of the degree of congestion. The study has failed to provide convincing and unambiguous data on the usefulness of examining mastoid processes and proximal attachments of the sternocleidomastoid muscles during autopsy to determine the presence of intravitality features of hanging. A description of research methodology and its associated difficulties, e.g. with the interpretation of results, can also be useful for the planning of similar studies by other researchers.

  14. Searching for new features of intravitality of hanging based on macro- and microscopic evaluation of the proximal attachment of the sternocleidomastoid muscle and the mastoid process of the temporal bone

    Directory of Open Access Journals (Sweden)

    Łukasz Szleszkowski

    2016-03-01

    Full Text Available Aim of the study : Assessment of the usefulness of intravital lesions in the proximal attachment of the sternocleidomastoid muscle and the mastoid process of the temporal bone in medico-legal evaluation of death by hanging. Material and methods: The study material was obtained from the bodies of 35 people who died by hanging. The control group comprised specimens collected from 30 people who died of non-traumatic causes. The structures under study were examined macro- and microscopically. The basic change which could be recognized as a marker of intravitality of hanging was the presence of a macroscopically extensive blotchy area of abundant ecchymosis in the proximal muscle attachment, similar to that found in the distal attachment, and the presence of abundant diffuse intraosseous ecchymoses in the mastoid process. Results: None of the cases revealed any ecchymoses in the proximal attachment of the muscle that would be similar to those present in the distal attachment. Discolourations within the mastoid processes, macroscopically suggestive of extensive intraosseous effusions arising from the mechanism of stretching, were not confirmed by microscopic evaluation and occurred at the same frequency as in the control group. Limitations of the study were related to the method which involved sample collection by means of bone chisels, decalcification and preparation of specimens, which had an effect, for example, on the measurable evaluation of the degree of congestion. Conclusions : The study has failed to provide convincing and unambiguous data on the usefulness of examining mastoid processes and proximal attachments of the sternocleidomastoid muscles during autopsy to determine the presence of intravitality features of hanging. A description of research methodology and its associated difficulties, e.g. with the interpretation of results, can also be useful for the planning of similar studies by other researchers.

  15. Multiphotonic Confocal Microscopy 3D imaging: Application to mantle sulfides in sub-arc environment (Avacha Volcano, Kamchatka)

    Science.gov (United States)

    Antoine, Bénard; Luc-Serge, Doucet; Sabine, Palle; Dmitri A., Ionov

    2010-05-01

    Petrogenetic relations in igneous rocks are usually studied in natural samples using classical optical microscopy and subsequent geochemical data acquisition. Multiphotonic Laser Scanning Confocal Microscopy (MLSCM) can be a powerful tool to section geological materials optically with sub-micrometric resolution and then generate a three-dimensional (3D) reconstruction (ca. 106 μm3 stack). MLSCM is used here to investigate textural relations of Monosulfide Solid Solution (MSS) with silicate phases in fresh spinel harzburgite xenoliths from the andesitic Avacha volcano (Kamchatka, Russia). The xenoliths contain MSS disseminated in olivine and orthopyroxene (opx) neoblasts as well as MSS-rich quenched magmatic opx veins [1]. First, Reflection Mode (RM) was tested on vein sulfides in resin-impregnated thick (120 μm) polished rock sections. Then we used a combination of Differential Interference Contrast (DIC) with a transmitted light detector, two photons-excited fluorescence (2PEF) and Second Harmonic Generation (SHG). Sequential imaging feature of the Leica TCS-SP2 software was applied. The excitation laser used for 2PEF was a COHERENT MIRA 900 with a 76Hz repetition rate and 800nm wavelength. Image stacks were analysed using ImageJ software [2]. The aim of the tests was to try to discriminate sulfides in silicate matrix as a tool for a better assessment of equilibrium conditions between the two phases. Preliminary results show that Fe-Ni rich MSS from vein and host rock have a strong auto-fluorescence in the Near UV-VIS domain (392-715 nm) whereas silicate matrix is only revealed through DIC. SHG is obtained only from dense nanocentrosymmetrical structures such as embedded medium (organic matter like glue and resin). The three images were recorded sequentially enabling efficient discrimination between the different components of the rock slices. RM permits reconstruction of the complete 3D structure of the rock slice. High resolution (ca. 0.2 μm along X-Y axis vs

  16. Advanced multiphoton methods for in vitro and in vivo functional imaging of mouse retinal neurons (Conference Presentation)

    Science.gov (United States)

    Cohen, Noam; Schejter, Adi; Farah, Nairouz; Shoham, Shy

    2016-03-01

    Studying the responses of retinal ganglion cell (RGC) populations has major significance in vision research. Multiphoton imaging of optogenetic probes has recently become the leading approach for visualizing neural populations and has specific advantages for imaging retinal activity during visual stimulation, because it leads to reduced direct photoreceptor excitation. However, multiphoton retinal activity imaging is not straightforward: point-by-point scanning leads to repeated neural excitation while optical access through the rodent eye in vivo has proven highly challenging. Here, we present two enabling optical designs for multiphoton imaging of responses to visual stimuli in mouse retinas expressing calcium indicators. First, we present an imaging solution based on Scanning Line Temporal Focusing (SLITE) for rapidly imaging neuronal activity in vitro. In this design, we scan a temporally focused line rather than a point, increasing the scan speed and reducing the impact of repeated excitation, while maintaining high optical sectioning. Second, we present the first in vivo demonstration of two-photon imaging of RGC activity in the mouse retina. To obtain these cellular resolution recordings we integrated an illumination path into a correction-free imaging system designed using an optical model of the mouse eye. This system can image at multiple depths using an electronically tunable lens integrated into its optical path. The new optical designs presented here overcome a number of outstanding obstacles, allowing the study of rapid calcium- and potentially even voltage-indicator signals both in vitro and in vivo, thereby bringing us a step closer toward distributed monitoring of action potentials.

  17. Resonance enhanced multiphoton ionization spectra of molecules and molecular fragments. Annual progress report, March 1992 - February 1993

    International Nuclear Information System (INIS)

    1993-01-01

    In this report, the author will review the progress made in his studies of ion rotational distributions resulting from resonance enhanced multiphoton ionization of excited electronic states and from single-photon ionization of ground electronic states of jet-cooled molecules by coherent VUV and XUV radiation. To do so he will select a few examples from his studies which serve to highlight his progress and to identify the background and significance of the specific spectral features and systems he has chosen to study

  18. Probing temporal aspects of high-order harmonic pulses via multi-colour, multi-photon ionization processes

    Energy Technology Data Exchange (ETDEWEB)

    Mauritsson, J [Department of Physics and Astronomy, Louisiana State University, Baton Rouge, LA 70803-4001 (United States); Johnsson, P [Department of Physics, Lund Institute of Technology, PO Box 118, SE-22100 Lund (Sweden); Lopez-Martens, R [Department of Physics, Lund Institute of Technology, PO Box 118, SE-22100 Lund (Sweden); Varju, K [Department of Physics, Lund Institute of Technology, PO Box 118, SE-22100 Lund (Sweden); L' Huillier, A [Department of Physics, Lund Institute of Technology, PO Box 118, SE-22100 Lund (Sweden); Gaarde, M B [Department of Physics and Astronomy, Louisiana State University, Baton Rouge, LA 70803-4001 (United States); Schafer, K J [Department of Physics and Astronomy, Louisiana State University, Baton Rouge, LA 70803-4001 (United States)

    2005-07-14

    High-order harmonics generated through the interaction of atoms and strong laser fields are a versatile, laboratory-scale source of extreme ultraviolet (XUV) radiation on a femtosecond or even attosecond time-scale. In order to be a useful experimental tool, however, this radiation has to be well characterized, both temporally and spectrally. In this paper we discuss how multi-photon, multi-colour ionization processes can be used to completely characterize either individual harmonics or attosecond pulse trains. In particular, we discuss the influence of the intensity and duration of the probe laser, and how these parameters effect the accuracy of the XUV characterization.

  19. Probing temporal aspects of high-order harmonic pulses via multi-colour, multi-photon ionization processes

    International Nuclear Information System (INIS)

    Mauritsson, J; Johnsson, P; Lopez-Martens, R; Varju, K; L'Huillier, A; Gaarde, M B; Schafer, K J

    2005-01-01

    High-order harmonics generated through the interaction of atoms and strong laser fields are a versatile, laboratory-scale source of extreme ultraviolet (XUV) radiation on a femtosecond or even attosecond time-scale. In order to be a useful experimental tool, however, this radiation has to be well characterized, both temporally and spectrally. In this paper we discuss how multi-photon, multi-colour ionization processes can be used to completely characterize either individual harmonics or attosecond pulse trains. In particular, we discuss the influence of the intensity and duration of the probe laser, and how these parameters effect the accuracy of the XUV characterization

  20. An Automated System for the Control of, and Data Acquisition from Multiphoton Ionization and Fluorescence Lifetime Measurements.

    Science.gov (United States)

    1986-09-01

    Quanta- Ray company , which also supplied the laser used for the multiphoton work. The, burner was mounted on a translator stage from Velmex, Inc...and no longer exists as a process in the system. When the user analysis program has completed, the lifetime program is again automatically re-started...KCHAR) RETURN 100 FORMAT(I3) 101 FORMAT(F7.2) END SUBROUTINE LAB4 FODA SE"oteD C This routine puts the label "INTEGRAL FROM DATA SET" on the MDP C screen

  1. Infrared Multiphoton Dissociation Spectroscopy with Free-Electron Lasers: On the Road from Small Molecules to Biomolecules.

    Science.gov (United States)

    Jašíková, Lucie; Roithová, Jana

    2018-03-07

    Infrared multiphoton dissociation (IRMPD) spectroscopy is commonly used to determine the structure of isolated, mass-selected ions in the gas phase. This method has been widely used since it became available at free-electron laser (FEL) user facilities. Thus, in this Minireview, we examine the use of IRMPD/FEL spectroscopy for investigating ions derived from small molecules, metal complexes, organometallic compounds and biorelevant ions. Furthermore, we outline new applications of IRMPD spectroscopy to study biomolecules. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Electronically tunable femtosecond all-fiber optical parametric oscillator for multi-photon microscopy

    Science.gov (United States)

    Hellwig, Tim; Brinkmann, Maximilian; Fallnich, Carsten

    2018-02-01

    We present a femtosecond fiber-based optical parametric oscillator (FOPO) for multiphoton microscopy with wavelength tuning by electronic repetition rate tuning in combination with a dispersive filter in the FOPO cavity. The all-spliced, all-fiber FOPO cavity is based on polarization-maintaining fibers and a broadband output coupler, allowing to get access to the resonant signal pulses as well as the idler pulses simultaneously. The system was pumped by a gain-switched fiber-coupled laser diode emitting pulses at a central wavelength of 1030 nm and an electronically tunable repetition frequency of about 2 MHz. The pump pulses were amplified in an Ytterbium fiber amplifier system with a pulse duration after amplification of 13 ps. Tuning of the idler (1140 nm - 1300 nm) and signal wavelengths (850 nm - 940 nm) was achieved by changing the repetition frequency of the pump laser by about 4 kHz. The generated signal pulses reached a pulse energy of up to 9.2 nJ at 920 nm and were spectrally broadened to about 6 nm in the FOPO by a combination of self-phase and cross-phase modulation. We showed external compression of the idler pulses at 920 nm to about 430 fs and appleid them to two-photon excitation microscopy with green fluorescent dyes. The presented system constitutes an important step towards a fully fiber-integrated all-electronically tunable and, thereby, programmable light source and already embodies a versatile and flexible light source for applications, e.g., for smart microscopy.

  3. Nonperturbative theory of single/multiphoton processes in atoms and molecules induced by intense laser fields

    International Nuclear Information System (INIS)

    Lau, A.M.F.

    1975-04-01

    A quantum nonperturbative theory is given for the problem of a general n discrete-level atomic/molecular system interacting with a strong single-mode/multimode radiation field. The atomic/molecular energy-level structures are modified due to interaction with the laser field. These energy level shifts are derived in the rigorous solution to the adiabatic eigenvalue problem of the charge--field system, involving a simple iterative procedure. The task of solution is simplified by recurrence relations between matrices connecting probability amplitudes of successive photon numbers. New formulae for calculating probability of single/multiphoton transitions between three resonant shifted levels and between some cases of two near-resonant shifted levels are derived. This general formalism can be applied to calculate transition probabilities of various atomic/molecular photo processes of interest. Numerical values are obtained for the inelastic cross section of the slow-collisional process Li + H and for dissociation cross section of LiH molecule. The transition probabilities of Na (3s → 5s by absorption of two photon of lambda = 0.60233μ -- 0.602396 μ) and of Li (2s → 3s by absorption of eight photons of lambda = 2.9406 μ -- 2.945 μ) irradiated by a strong pulse are calculated. Finally, a parametric study is carried out for the process where a molecular system is interacting with two intense radiation fields of different wavelengths. Owing to potential barrier shift due to the much more intense field, the molecular system penetrates into an otherwise inaccessible region in the potential level where it is allowed to radiate to a lower level by emitting photons at a second wavelength. (12 figures, 6 tables) (U.S.)

  4. Photo-induced changes of silicate glasses optical parameters at multi-photon laser radiation absorption

    International Nuclear Information System (INIS)

    Efimov, O.M.; Glebov, L.B.; Mekryukov, A.M.

    1995-01-01

    In this paper the results of investigations of the mechanisms of photo-induced changes of alkali-silicate (crown) and lead-silicate (flint) glasses optical parameters upon the exposure to the intense laser radiation, and the basic regularities of these processes are reported. These investigations were performed in Research Center open-quotes S. I. Vavilov State Optical Instituteclose quotes during last 15 years. The kinetics of stable and unstable CC formation and decay, the effect of widely spread impurity ions on these processes, the characteristics of fundamental and impure luminescence, the kinetics of refractive index change under conditions of multi-photon glass matrix excitation, and other properties are considered. On the basis of analysis of received regularities it was shown that the nonlinear coloration of alkali-silicate glasses (the fundamental absorption edge is nearly 6 eV) takes place only as a result of two-photon absorption. Important efforts were aimed at the detection of three- or more photon matrix ionization of these glasses, but they were failed. However it was established that in the lead silicate glasses the long-wave carriers mobility boundary (> 5.6 eV) is placed considerably higher the fundamental absorption edge (∼ 3.5 eV) of material matrix. This results in that the linear color centers formation in the lead silicate glasses is not observed. The coloration of these glasses arises only from the two- or three-photon matrix ionization, and the excitation occurs through virtual states that are placed in the fundamental absorption region. In the report the available mechanisms of photo-induced changes of glasses optical parameters, and some applied aspects of this problem are discussed

  5. Examination of wound healing after curettage by multiphoton tomography of human skin in vivo.

    Science.gov (United States)

    Springer, S; Zieger, M; Böttcher, A; Lademann, J; Kaatz, M

    2017-11-01

    The multiphoton tomography (MPT) has evolved into a useful tool for the non-invasive investigation of morphological and biophysical characteristics of human skin in vivo. Until now, changes of the skin have been evaluated mainly by using clinical and histological techniques. In this study, the progress of wound healing was investigated by MPT over 3 weeks with a final examination after 24 months. Especially, the collagen degradation, reepithelization and tissue formation were examined. As specific parameter for wound healing and its course the second-harmonic generation-to-autofluorescence aging index of dermis (SAAID) was used. About 10 volunteers aged between 25 and 58 years were examined. Acute wounds were scanned with three Z-stacks taken per visit. The stacks were taken up to a depth of 225 μm at increments of 5 μm and a scan time for 3 seconds per scan. Subsequently, the SAAID was evaluated as an indicator for wound healing. Furthermore, single scans were taken for morphological investigations. The evaluation revealed a distinct difference in the SAAID behavior between the Z-stacks taken at each visit. Furthermore, the degradation of collagen and cells and their reappearance could be shown in the course of the visits. Clear differences in the curve behavior of the SAAID at every visit were shown in this study. The SAAID curves and morphological images could be correlated with findings of the clinical examination of different wound healing phases. Therefore, SAAID curves and morphological MPT imaging could provide a non-invasive tool for the determination of wound healing phases in patients in vivo. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  6. Comparison of in vivo and ex vivo laser scanning microscopy and multiphoton tomography application for human and porcine skin imaging

    Energy Technology Data Exchange (ETDEWEB)

    Darvin, M E; Richter, H; Zhu, Y J; Meinke, M C; Knorr, F; Lademann, J [Center of Experimental and Applied Cutaneous Physiology, Department of Dermatology, Venerology and Allergology, Charité - Universitätsmedizin Berlin (Germany); Gonchukov, S A [National Research Nuclear University ' ' MEPhI' ' (Russian Federation); Koenig, K [JenLab GmbH, Schillerstr. 1, 07745 Jena (Germany)

    2014-07-31

    Two state-of-the-art microscopic optical methods, namely, confocal laser scanning microscopy in the fluorescence and reflectance regimes and multiphoton tomography in the autofluorescence and second harmonic generation regimes, are compared for porcine skin ex vivo and healthy human skin in vivo. All skin layers such as stratum corneum (SC), stratum spinosum (SS), stratum basale (SB), papillary dermis (PD) and reticular dermis (RD) as well as transition zones between these skin layers are measured noninvasively at a high resolution, using the above mentioned microscopic methods. In the case of confocal laser scanning microscopy (CLSM), measurements in the fluorescence regime were performed by using a fluorescent dye whose topical application on the surface is well suited for the investigation of superficial SC and characterisation of the skin barrier function. For investigations of deeply located skin layers, such as SS, SB and PD, the fluorescent dye must be injected into the skin, which markedly limits fluorescence measurements using CLSM. In the case of reflection CLSM measurements, the obtained results can be compared to the results of multiphoton tomography (MPT) for all skin layers excluding RD. CLSM cannot distinguish between dermal collagen and elastin measuring their superposition in the RD. By using MPT, it is possible to analyse the collagen and elastin structures separately, which is important for the investigation of anti-aging processes. The resolution of MPT is superior to CLSM. The advantages and limitations of both methods are discussed and the differences and similarities between human and porcine skin are highlighted. (laser biophotonics)

  7. Multi-photon resonant effects in strong-field ionization: origin of the dip in experimental longitudinal momentum distributions

    International Nuclear Information System (INIS)

    Alnaser, A S; Maharjan, C M; Wang, P; Litvinyuk, I V

    2006-01-01

    We studied ionization of neon and argon by intense linearly polarized femtosecond laser pulses of different wavelengths (400 nm and 800 nm) and peak intensities, and by measuring momentum distributions of singly charged positive ions in the direction parallel to laser polarization. For Ne the momentum distributions exhibited a characteristic dip at zero momentum at 800 nm and a complex multipeak structure at 400 nm. Similarly, for Ar the momentum distributions evolved from a complex multipeak structure with a pronounced dip in the centre at 400 nm, to a smooth distribution characteristic of pure tunneling ionization (800 nm, high intensities). In the intermediate regime (800 nm, medium to low intensities), for both atoms we observed recoil ion momentum distributions modulated by quasi-periodic structures usually seen in the photoelectron energy spectra in a multi-photon regime (ATI spectra). Ne did show a characteristic 'dip' at low momentum, while the longitudinal momentum distribution for Ar exhibited a spike at zero momentum instead. The spectra did dramatically change at 400 nm, where both ions show the pronounced dip near zero momentum. Based on our results, we conclude that the structures observed in Ne and Ar momentum distributions reflect the specifics of atomic structure of the two targets and should not be attributed to effects of electron recollision, as was suggested earlier. Instead, as our results indicate, they are due to the effects of multi-photon resonant enhancement of strong-field ionization. (letter to the editor)

  8. Comparison of in vivo and ex vivo laser scanning microscopy and multiphoton tomography application for human and porcine skin imaging

    Science.gov (United States)

    Darvin, M. E.; Richter, H.; Zhu, Y. J.; Meinke, M. C.; Knorr, F.; Gonchukov, S. A.; Koenig, K.; Lademann, J.

    2014-07-01

    Two state-of-the-art microscopic optical methods, namely, confocal laser scanning microscopy in the fluorescence and reflectance regimes and multiphoton tomography in the autofluorescence and second harmonic generation regimes, are compared for porcine skin ex vivo and healthy human skin in vivo. All skin layers such as stratum corneum (SC), stratum spinosum (SS), stratum basale (SB), papillary dermis (PD) and reticular dermis (RD) as well as transition zones between these skin layers are measured noninvasively at a high resolution, using the above mentioned microscopic methods. In the case of confocal laser scanning microscopy (CLSM), measurements in the fluorescence regime were performed by using a fluorescent dye whose topical application on the surface is well suited for the investigation of superficial SC and characterisation of the skin barrier function. For investigations of deeply located skin layers, such as SS, SB and PD, the fluorescent dye must be injected into the skin, which markedly limits fluorescence measurements using CLSM. In the case of reflection CLSM measurements, the obtained results can be compared to the results of multiphoton tomography (MPT) for all skin layers excluding RD. CLSM cannot distinguish between dermal collagen and elastin measuring their superposition in the RD. By using MPT, it is possible to analyse the collagen and elastin structures separately, which is important for the investigation of anti-aging processes. The resolution of MPT is superior to CLSM. The advantages and limitations of both methods are discussed and the differences and similarities between human and porcine skin are highlighted.

  9. Comparison of in vivo and ex vivo laser scanning microscopy and multiphoton tomography application for human and porcine skin imaging

    International Nuclear Information System (INIS)

    Darvin, M E; Richter, H; Zhu, Y J; Meinke, M C; Knorr, F; Lademann, J; Gonchukov, S A; Koenig, K

    2014-01-01

    Two state-of-the-art microscopic optical methods, namely, confocal laser scanning microscopy in the fluorescence and reflectance regimes and multiphoton tomography in the autofluorescence and second harmonic generation regimes, are compared for porcine skin ex vivo and healthy human skin in vivo. All skin layers such as stratum corneum (SC), stratum spinosum (SS), stratum basale (SB), papillary dermis (PD) and reticular dermis (RD) as well as transition zones between these skin layers are measured noninvasively at a high resolution, using the above mentioned microscopic methods. In the case of confocal laser scanning microscopy (CLSM), measurements in the fluorescence regime were performed by using a fluorescent dye whose topical application on the surface is well suited for the investigation of superficial SC and characterisation of the skin barrier function. For investigations of deeply located skin layers, such as SS, SB and PD, the fluorescent dye must be injected into the skin, which markedly limits fluorescence measurements using CLSM. In the case of reflection CLSM measurements, the obtained results can be compared to the results of multiphoton tomography (MPT) for all skin layers excluding RD. CLSM cannot distinguish between dermal collagen and elastin measuring their superposition in the RD. By using MPT, it is possible to analyse the collagen and elastin structures separately, which is important for the investigation of anti-aging processes. The resolution of MPT is superior to CLSM. The advantages and limitations of both methods are discussed and the differences and similarities between human and porcine skin are highlighted. (laser biophotonics)

  10. Miniature fiber-optic multiphoton microscopy system using frequency-doubled femtosecond Er-doped fiber laser.

    Science.gov (United States)

    Huang, Lin; Mills, Arthur K; Zhao, Yuan; Jones, David J; Tang, Shuo

    2016-05-01

    We report on a miniature fiber-optic multiphoton microscopy (MPM) system based on a frequency-doubled femtosecond Er-doped fiber laser. The femtosecond pulses from the laser source are delivered to the miniature fiber-optic probe at 1.58 µm wavelength, where a standard single mode fiber is used for delivery without the need of free-space dispersion compensation components. The beam is frequency-doubled inside the probe by a periodically poled MgO:LiNbO3 crystal. Frequency-doubled pulses at 786 nm with a maximum power of 80 mW and a pulsewidth of 150 fs are obtained and applied to excite intrinsic signals from tissues. A MEMS scanner, a miniature objective, and a multimode collection fiber are further used to make the probe compact. The miniature fiber-optic MPM system is highly portable and robust. Ex vivo multiphoton imaging of mammalian skins demonstrates the capability of the system in imaging biological tissues. The results show that the miniature fiber-optic MPM system using frequency-doubled femtosecond fiber laser can potentially bring the MPM imaging for clinical applications.

  11. Multiphoton Absorption Order of CsPbBr3 As Determined by Wavelength-Dependent Nonlinear Optical Spectroscopy.

    Science.gov (United States)

    Saouma, Felix O; Stoumpos, Constantinos C; Kanatzidis, Mercouri G; Kim, Yong Soo; Jang, Joon I

    2017-10-05

    CsPbBr 3 is a direct-gap semiconductor where optical absorption takes place across the fundamental bandgap, but this all-inorganic halide perovskite typically exhibits above-bandgap emission when excited over an energy level, lying above the conduction-band minimum. We probe this bandgap anomaly using wavelength-dependent multiphoton absorption spectroscopy and find that the fundamental gap is strictly two-photon forbidden, rendering it three-photon absorption (3PA) active. Instead, two-photon absorption (2PA) commences when the two-photon energy is resonant with the optical gap, associated with the level causing the anomaly. We determine absolute nonlinear optical dispersion over this 3PA-2PA region, which can be explained by two-band models in terms of the optical gap. The polarization dependence of 3PA and 2PA is also measured and explained by the relevant selection rules. CsPbBr 3 is highly luminescent under multiphoton absorption at room temperature with marked polarization and wavelength dependence at the 3PA-2PA crossover and therefore has potential for nonlinear optical applications.

  12. Single and multi-photon events with missing energy in $e^+ e^-$ collisions at 161 GeV < $\\sqrt{s}$ < 172 GeV

    CERN Document Server

    Acciarri, M; Aguilar-Benítez, M; Ahlen, S P; Alcaraz, J; Alemanni, G; Allaby, James V; Aloisio, A; Alverson, G; Alviggi, M G; Ambrosi, G; Anderhub, H; Andreev, V P; Angelescu, T; Anselmo, F; Arefev, A; Azemoon, T; Aziz, T; Bagnaia, P; Baksay, L; Banerjee, S; Banerjee, Sw; Banicz, K; Barczyk, A; Barillère, R; Barone, L; Bartalini, P; Baschirotto, A; Basile, M; Battiston, R; Bay, A; Becattini, F; Becker, U; Behner, F; Berdugo, J; Berges, P; Bertucci, B; Betev, B L; Bhattacharya, S; Biasini, M; Biland, A; Bilei, G M; Blaising, J J; Blyth, S C; Bobbink, Gerjan J; Böck, R K; Böhm, A; Boldizsar, L; Borgia, B; Bourilkov, D; Bourquin, Maurice; Braccini, S; Branson, J G; Brigljevic, V; Brock, I C; Buffini, A; Buijs, A; Burger, J D; Burger, W J; Busenitz, J K; Button, A M; Cai, X D; Campanelli, M; Capell, M; Cara Romeo, G; Carlino, G; Cartacci, A M; Casaus, J; Castellini, G; Cavallari, F; Cavallo, N; Cecchi, C; Cerrada-Canales, M; Cesaroni, F; Chamizo-Llatas, M; Chang, Y H; Chaturvedi, U K; Chekanov, S V; Chemarin, M; Chen, A; Chen, G; Chen, G M; Chen, H F; Chen, H S; Chéreau, X J; Chiefari, G; Chien, C Y; Cifarelli, Luisa; Cindolo, F; Civinini, C; Clare, I; Clare, R; Cohn, H O; Coignet, G; Colijn, A P; Colino, N; Commichau, V; Costantini, S; Cotorobai, F; de la Cruz, B; Csilling, Akos; Dai, T S; D'Alessandro, R; De Asmundis, R; Degré, A; Deiters, K; Della Volpe, D; Denes, P; De Notaristefani, F; DiBitonto, Daryl; Diemoz, M; Van Dierendonck, D N; Di Lodovico, F; Dionisi, C; Dittmar, Michael; Dominguez, A; Doria, A; Dova, M T; Duchesneau, D; Duinker, P; Durán, I; Dutta, S; Easo, S; Efremenko, Yu V; El-Mamouni, H; Engler, A; Eppling, F J; Erné, F C; Ernenwein, J P; Extermann, Pierre; Fabre, M; Faccini, R; Falciano, S; Favara, A; Fay, J; Fedin, O; Felcini, Marta; Fenyi, B; Ferguson, T; Ferroni, F; Fesefeldt, H S; Fiandrini, E; Field, J H; Filthaut, Frank; Fisher, P H; Fisk, I; Forconi, G; Fredj, L; Freudenreich, Klaus; Furetta, C; Galaktionov, Yu; Ganguli, S N; García-Abia, P; Gau, S S; Gentile, S; Gheordanescu, N; Giagu, S; Goldfarb, S; Goldstein, J; Gong, Z F; Gougas, Andreas; Gratta, Giorgio; Grünewald, M W; Gupta, V K; Gurtu, A; Gutay, L J; Hartmann, B; Hasan, A; Hatzifotiadou, D; Hebbeker, T; Hervé, A; Van Hoek, W C; Hofer, H; Hong, S J; Hoorani, H; Hou, S R; Hu, G; Innocente, Vincenzo; Jenkes, K; Jin, B N; Jones, L W; de Jong, P; Josa-Mutuberria, I; Kasser, A; Khan, R A; Kamrad, D; Kamyshkov, Yu A; Kapustinsky, J S; Karyotakis, Yu; Kaur, M; Kienzle-Focacci, M N; Kim, D; Kim, D H; Kim, J K; Kim, S C; Kim, Y G; Kinnison, W W; Kirkby, A; Kirkby, D; Kirkby, Jasper; Kiss, D; Kittel, E W; Klimentov, A; König, A C; Kopp, A; Korolko, I; Koutsenko, V F; Krämer, R W; Krenz, W; Kunin, A; Ladrón de Guevara, P; Laktineh, I; Landi, G; Lapoint, C; Lassila-Perini, K M; Laurikainen, P; Lebeau, M; Lebedev, A; Lebrun, P; Lecomte, P; Lecoq, P; Le Coultre, P; Le Goff, J M; Leiste, R; Leonardi, E; Levchenko, P M; Li Chuan; Lin, C H; Lin, W T; Linde, Frank L; Lista, L; Liu, Z A; Lohmann, W; Longo, E; Lu, W; Lü, Y S; Lübelsmeyer, K; Luci, C; Luckey, D; Luminari, L; Lustermann, W; Ma Wen Gan; Maity, M; Majumder, G; Malgeri, L; Malinin, A; Maña, C; Mangeol, D J J; Mangla, S; Marchesini, P A; Marin, A; Martin, J P; Marzano, F; Massaro, G G G; McNally, D; McNeil, R R; Mele, S; Merola, L; Meschini, M; Metzger, W J; Von der Mey, M; Mi, Y; Mihul, A; Van Mil, A J W; Mirabelli, G; Mnich, J; Molnár, P; Monteleoni, B; Moore, R; Morganti, S; Moulik, T; Mount, R; Müller, S; Muheim, F; Muijs, A J M; Nahn, S; Napolitano, M; Nessi-Tedaldi, F; Newman, H; Niessen, T; Nippe, A; Nisati, A; Nowak, H; Oh, Yu D; Opitz, H; Organtini, G; Ostonen, R; Palomares, C; Pandoulas, D; Paoletti, S; Paolucci, P; Park, H K; Park, I H; Pascale, G; Passaleva, G; Patricelli, S; Paul, T; Pauluzzi, M; Paus, C; Pauss, Felicitas; Peach, D; Pei, Y J; Pensotti, S; Perret-Gallix, D; Petersen, B; Petrak, S; Pevsner, A; Piccolo, D; Pieri, M; Pinto, J C; Piroué, P A; Pistolesi, E; Plyaskin, V; Pohl, M; Pozhidaev, V; Postema, H; Produit, N; Prokofev, D; Prokofiev, D O; Rahal-Callot, G; Raja, N; Rancoita, P G; Rattaggi, M; Raven, G; Razis, P A; Read, K; Ren, D; Rescigno, M; Reucroft, S; Van Rhee, T; Riemann, S; Riles, K; Robohm, A; Rodin, J; Roe, B P; Romero, L; Rosier-Lees, S; Rosselet, P; Van Rossum, W; Roth, S; Rubio, Juan Antonio; Ruschmeier, D; Rykaczewski, H; Salicio, J; Sánchez, E; Sanders, M P; Sarakinos, M E; Sarkar, S; Sassowsky, M; Schäfer, C; Shchegelskii, V; Schmidt-Kärst, S; Schmitz, D; Schmitz, P; Scholz, N; Schopper, Herwig Franz; Schotanus, D J; Schwenke, J; Schwering, G; Sciacca, C; Sciarrino, D; Servoli, L; Shevchenko, S; Shivarov, N; Shoutko, V; Shukla, J; Shumilov, E; Shvorob, A V; Siedenburg, T; Son, D; Sopczak, André; Smith, B; Spillantini, P; Steuer, M; Stickland, D P; Stone, A; Stone, H; Stoyanov, B; Strässner, A; Strauch, K; Sudhakar, K; Sultanov, G G; Sun, L Z; Susinno, G F; Suter, H; Swain, J D; Tang, X W; Tauscher, Ludwig; Taylor, L; Ting, Samuel C C; Ting, S M; Tonutti, M; Tonwar, S C; Tóth, J; Tully, C; Tuchscherer, H; Tung, K L; Uchida, Y; Ulbricht, J; Uwer, U; Valente, E; Van de Walle, R T; Vesztergombi, G; Vetlitskii, I; Viertel, Gert M; Vivargent, M; Völkert, R; Vogel, H; Vogt, H; Vorobev, I; Vorobyov, A A; Vorvolakos, A; Wadhwa, M; Wallraff, W; Wang, J C; Wang, X L; Wang, Z M; Weber, A; Wittgenstein, F; Wu, S X; Wynhoff, S; Xu, J; Xu, Z Z; Yang, B Z; Yang, C G; Yao, X Y; Ye, J B; Yeh, S C; You, J M; Zalite, A; Zalite, Yu; Zemp, P; Zeng, Y; Zhang, Z; Zhang, Z P; Zhou, B; Zhu, G Y; Zhu, R Y; Zichichi, Antonino; Ziegler, F

    1997-01-01

    A search for single and multi-photon events with missing energy is performed using data collected at centre-of-mass energies between 161 GeV and 172 GeV for a total of 20.9 pb$^{-1}$ of integrated luminosity. The results obtained are used to derive the value for the $\

  13. In vivo multiphoton-microscopy of picosecond-laser-induced optical breakdown in human skin.

    Science.gov (United States)

    Balu, Mihaela; Lentsch, Griffin; Korta, Dorota Z; König, Karsten; Kelly, Kristen M; Tromberg, Bruce J; Zachary, Christopher B

    2017-08-01

    Improvements in skin appearance resulting from treatment with fractionated picosecond-lasers have been noted, but optimizing the treatment efficacy depends on a thorough understanding of the specific skin response. The development of non-invasive laser imaging techniques in conjunction with laser therapy can potentially provide feedback for guidance and optimizing clinical outcome. The purpose of this study was to demonstrate the capability of multiphoton microscopy (MPM), a high-resolution, label-free imaging technique, to characterize in vivo the skin response to a fractionated non-ablative picosecond-laser treatment. Two areas on the arm of a volunteer were treated with a fractionated picosecond laser at the Dermatology Clinic, UC Irvine. The skin response to treatment was imaged in vivo with a clinical MPM-based tomograph at 3 hours and 24 hours after treatment and seven additional time points over a 4-week period. MPM revealed micro-injuries present in the epidermis. Pigmented cells were particularly damaged in the process, suggesting that melanin is likely the main absorber for laser induced optical breakdown. Damaged individual cells were distinguished as early as 3 hours post pico-laser treatment with the 532 nm wavelength, and 24 hours post-treatment with both 532 and 1064 nm wavelengths. At later time points, clusters of cellular necrotic debris were imaged across the treated epidermis. After 24 hours of treatment, inflammatory cells were imaged in the proximity of epidermal micro-injuries. The epidermal injuries were exfoliated over a 4-week period. This observational and descriptive pilot study demonstrates that in vivo MPM imaging can be used non-invasively to provide label-free contrast for describing changes in human skin following a fractionated non-ablative laser treatment. The results presented in this study represent the groundwork for future longitudinal investigations on an expanded number of subjects to understand the response to treatment

  14. A novel flexible clinical multiphoton tomograph for early melanoma detection, skin analysis, testing of anti-age products, and in situ nanoparticle tracking

    Science.gov (United States)

    Weinigel, Martin; Breunig, Hans Georg; Gregory, Axel; Fischer, Peter; Kellner-Höfer, Marcel; Bückle, Rainer; König, Karsten

    2010-02-01

    High-resolution 3D microscopy based on multiphoton induced autofluorescence and second harmonic generation have been introduced in 1990. 13 years later, CE-marked clinical multiphoton systems for 3D imaging of human skin with subcellular resolution have first been launched by JenLab company with the tomography DermaInspect®. This year, the second generation of clinical multiphoton tomographs was introduced. The novel multiphoton tomograph MPTflex, equipped with a flexible articulated optical arm, provides an increased flexibility and accessibility especially for clinical and cosmetical examinations. Improved image quality and signal to noise ratio (SNR) are achieved by a very short source-drain spacing, by larger active areas of the detectors and by single photon counting (SPC) technology. Shorter image acquisition time due to improved image quality reduces artifacts and simplifies the operation of the system. The compact folded optical design and the light-weight structure of the optical head eases the handling. Dual channel detectors enable to distinguish between intratissue elastic fibers and collagenous structures simultaneously. Through the use of piezo-driven optics a stack of optical cross-sections (optical sectioning) can be acquired and 3D imaging can be performed. The multiphoton excitation of biomolecules like NAD(P)H, flavins, porphyrins, elastin, and melanin is done by picojoule femtosecond laser pulses from an tunable turn-key femtosescond near infrared laser system. The ability for rapid high-quality image acquisition, the user-friendly operation of the system and the compact and flexible design qualifies this system to be used for melanoma detection, diagnostics of dermatological disorders, cosmetic research and skin aging measurements as well as in situ drug monitoring and animal research.

  15. Wavelength dependent photoelectron circular dichroism of limonene studied by femtosecond multiphoton laser ionization and electron-ion coincidence imaging

    Science.gov (United States)

    Rafiee Fanood, Mohammad M.; Janssen, Maurice H. M.; Powis, Ivan

    2016-09-01

    Enantiomers of the monoterpene limonene have been investigated by (2 + 1) resonance enhanced multiphoton ionization and photoelectron circular dichroism employing tuneable, circularly polarized femtosecond laser pulses. Electron imaging detection provides 3D momentum measurement while electron-ion coincidence detection can be used to mass-tag individual electrons. Additional filtering, by accepting only parent ion tagged electrons, can be then used to provide discrimination against higher energy dissociative ionization mechanisms where more than three photons are absorbed to better delineate the two photon resonant, one photon ionization pathway. The promotion of different vibrational levels and, tentatively, different electronic ion core configurations in the intermediate Rydberg states can be achieved with different laser excitation wavelengths (420 nm, 412 nm, and 392 nm), in turn producing different state distributions in the resulting cations. Strong chiral asymmetries in the lab frame photoelectron angular distributions are quantified, and a comparison made with a single photon (synchrotron radiation) measurement at an equivalent photon energy.

  16. Status of FEL-SUT project, and the experimental setup for multiphoton dissociation and isotope separation in the gaseous phase

    CERN Document Server

    Chernyshev, A V; Petrov, A K; Kawai, M; Toyoda, K; Nakai, K; Kuroda, H

    2001-01-01

    The IR FEL Research Center of the Science University of Tokyo (FEL-SUT) is open for users to develop new applications of IR FEL in a wide field of material science, chemical technology and bio-chemical applications. The FEL is based on 35 MeV linac operated at the frequency of 2856 MHz (s-band). The FEL covers the wavelength range from 5 to 16 mu m with the micropulse duration of 1-2 ps, macropulse duration of 1 mu s, macropulse repetition rate of 10 Hz and the overall average power of 1 W. We report the present status of the Center and an experimental setup designed and constructed for the experiments on multiphoton dissociation and isotope separation.

  17. In vivo, label-free, three-dimensional quantitative imaging of liver surface using multi-photon microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Zhuo, Shuangmu, E-mail: shuangmuzhuo@gmail.com, E-mail: hanry-yu@nuhs.edu.sg [Biosystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #04-13/14 Enterprise Wing, 138602 Singapore (Singapore); Institute of Laser and Optoelectronics Technology, Fujian Normal University, Fuzhou 350007 (China); Yan, Jie [Biosystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #04-13/14 Enterprise Wing, 138602 Singapore (Singapore); Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, #04-01, 138669 Singapore (Singapore); Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, 14 Medical Drive, MD 11 #04-01A, 117599 Singapore (Singapore); Kang, Yuzhan [Biosystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #04-13/14 Enterprise Wing, 138602 Singapore (Singapore); Xu, Shuoyu [Biosystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #04-13/14 Enterprise Wing, 138602 Singapore (Singapore); Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, #04-01, 138669 Singapore (Singapore); Computation and System Biology Program, Singapore-MIT Alliance, 4 Engineering Drive 3, E4-04-10, 117576 Singapore (Singapore); Peng, Qiwen [Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, #04-01, 138669 Singapore (Singapore); Computation and System Biology Program, Singapore-MIT Alliance, 4 Engineering Drive 3, E4-04-10, 117576 Singapore (Singapore); Mechanobiology Institute, 5A Engineering Drive 1, T-Lab #05-01, 117411 Singapore (Singapore); and others

    2014-07-14

    Various structural features on the liver surface reflect functional changes in the liver. The visualization of these surface features with molecular specificity is of particular relevance to understanding the physiology and diseases of the liver. Using multi-photon microscopy (MPM), we have developed a label-free, three-dimensional quantitative and sensitive method to visualize various structural features of liver surface in living rat. MPM could quantitatively image the microstructural features of liver surface with respect to the sinuosity of collagen fiber, the elastic fiber structure, the ratio between elastin and collagen, collagen content, and the metabolic state of the hepatocytes that are correlative with the pathophysiologically induced changes in the regions of interest. This study highlights the potential of this technique as a useful tool for pathophysiological studies and possible diagnosis of the liver diseases with further development.

  18. Multiphoton ionization of (Xe)/sub n/ and (NO)/sub n/ clusters using a picosecond laser

    International Nuclear Information System (INIS)

    Smith, D.B.; Miller, J.C.

    1989-01-01

    In an effort to extend the application of multiphoton ionization (MPI) spectroscopy to the study of weakly bound systems, we have begun a systematic investigation of picosecond MPI in van der Waals molecules and clusters. To our knowledge no previous picosecond MPI studies of weakly bound systems have been reported. We present here results of picosecond MPI of Xe/sub n/(n = 1-20) and (NO)/sub n/(n = 1-4) clusters. Previous MPI studies using nanosecond lasers have not detected the NO cluster series, presumably because of fast dissociation channels. The use of high peak-power allows resonant and non-resonant photon absorption to the ionization limit to compete effectively with fast dissociative processes. 10 refs., 2 figs

  19. Multiphoton microscopic imaging of histological sections without hematoxylin and eosin staining differentiates carcinoma in situ lesion from normal oesophagus

    Science.gov (United States)

    Chen, Jianxin; Xu, Jian; Kang, Deyong; Xu, Meifang; Zhuo, Shuangmu; Zhu, Xiaoqin; Jiang, Xingshan

    2013-10-01

    Multiphoton microscopy (MPM) has become a powerful, important tool for tissues imaging at the molecular level. In this paper, this technique was extended to histological investigations, differentiating carcinoma in situ (CIS) lesion from normal oesophagus by imaging histological sections without hematoxylin and eosin (H&E) staining. The results show that the histology procedures of dehydration, paraffin embedding, and de-paraffinizing highlighted two photon excited fluorescence of cytoplasm and nucleolus of epithelial cell and collagen in stroma. MPM has the ability to identify the characteristics of CIS lesion including changes of squamous cells and full epithelium, identification of basement membrane, especially prominent nucleolus. The studies described here show that MPM has the potential for future retrospective studies of tumor staging by employing on histological section specimens without H&E staining.

  20. Theory of the multiphoton cascade transitions with two photon links: comparison of quantum electrodynamical and quantum mechanical approaches

    International Nuclear Information System (INIS)

    Zalialiutdinov, T; Baukina, Yu; Solovyev, D; Labzowsky, L

    2014-01-01

    The theory of multiphoton cascade transitions with two-photon links is considered within two different approaches: quantum electrodynamical (QED) and phenomenological quantum mechanical (QM). A problem of regularization of the cascade contributions is investigated in detail. It is argued that the correct regularization should include both initial and intermediate level widths in the singular energy denominators. This result follows both from the QED and from the QM approach. Particular transitions nl → 1s + 2γ with nl = 3s, 4s, 3d, 4d and nl → 1s + 3γ with nl = 3p, 4p are considered as examples. The importance of the proper cascade regularization is also demonstrated. (paper)

  1. On the necessity of taking into account the contribution of multiphoton exchanges into electron-proton deep inelastic scattering

    International Nuclear Information System (INIS)

    Savrin, V.I.

    1979-01-01

    The hypothesis that the multiphoton exchanges give a substantial contribution to the electron-proton inclusive scattering is formulated. The hypothesis explains the observed violation of the Bjorken scaling law. As it is shown, the mechanism of such intensification of multiple exchanges may by connected with the properties of the processes of hadron multiproduction in the deep inelastic field. This results in the necessity to calculate the inclusive cross section in all electromagnetic coupling constant orders. This has been done in the framework of the density matrix method. As a result the deep inelastic scattering cross section calculated without application of the perturbation theory reveals a new property of the scaling invariance and leads to the natural relationship of structural functions with electromagnetic proton form-factors on the exclusive threshold

  2. Multiphoton control of the 1,3-cyclohexadiene ring-opening reaction in the presence of competing solvent reactions.

    Science.gov (United States)

    Carroll, Elizabeth C; White, James L; Florean, Andrei C; Bucksbaum, Philip H; Sension, Roseanne J

    2008-07-31

    Although physical chemistry has often concentrated on the observation and understanding of chemical systems, the defining characteristic of chemistry remains the direction and control of chemical reactivity. Optical control of molecular dynamics, and thus of chemical reactivity provides a path to use photon energy as a smart reagent in a chemical system. In this paper, we discuss recent research in this field in the context of our studies of the multiphoton optical control of the photo-initiated ring-opening reaction of 1,3-cyclohexadiene (CHD) to form 1,3,5- cis-hexatriene (Z-HT). Closed-loop feedback and learning algorithms are able to identify pulses that increase the desired target state by as much as a factor of two. Mechanisms for control are discussed through the influence of the intensity dependence, the nonlinear power spectrum, and the projection of the pulses onto low orders of polynomial phase. Control measurements in neat solvents demonstrate that competing solvent fragmentation reactions must also be considered. In particular, multiphoton excitation of cyclohexane alone is capable of producing hexatriene. Statistical analyses of data sets obtained in learning algorithm searches in neat cyclohexane and for CHD in hexane and cyclohexane highlight the importance of linear and quadratic chirp, while demonstrating that the control features are not so easily defined. Higher order phase components are also important. On the basis of these results the involvement of low-frequency ground-state vibrational modes is proposed. When the population is transferred to the excited state, momentum along the torsional coordinate may keep the wave packet localized as it moves toward the conical intersections controlling the yield of Z-HT.

  3. Theory of the effect of third-harmonic generation on three-photon resonantly enhanced multiphoton ionization in focused beams

    International Nuclear Information System (INIS)

    Payne, M.G.; Garrett, W.R.

    1983-01-01

    Multiphoton ionization in the region near a three-photon resonance is treated for focused, plane-polarized Gaussian beams with diffraction-limited beam divergence. In this situation, a third-harmonic field is generated within the laser beam. At, and very near, three-photon resonance the driving rate for the upper-state probability amplitude due to one-photon absorption of third-harmonic light becomes nearly equal to the corresponding three-photon rate due to the laser field, but these effects are 180 0 out of phase. As a consequence of this cancellation between two pumping terms, the three-photon resonance line essentially disappears at moderate concentrations and the observed ionization has a line shape that is close to the phase-matching curve for third-harmonic generation. The ionization signal, near but not on the resonance, is due almost entirely to absorption of third-harmonic photons plus other laser photons; three-photon resonantly enhanced multiphoton ionization by the laser is much weaker. This is particularly true on the blue side of the three-photon resonance at detunings where phase matching occurs. The problem is treated quite generally with predictions of the full line shape for n-photon ionization and third-harmonic light generation near three-photon resonance, including the rather strong influences of positively dispersive buffer gases. We also show that the cancellation between the one-photon and the three-photon process is partially spoiled in the presence of a counterpropagating beam at the same frequency

  4. Simulation of multi-photon emission isotopes using time-resolved SimSET multiple photon history generator

    Energy Technology Data Exchange (ETDEWEB)

    Chiang, Chih-Chieh; Lin, Hsin-Hon; Lin, Chang-Shiun; Chuang, Keh-Shih [Department of Biomedical Engineering and Environmental Sciences, National Tsing-HuaUniversity, Hsinchu, Taiwan (China); Jan, Meei-Ling [Health Physics Division, Institute of Nuclear Energy Research, Atomic Energy Council, Taoyuan, Taiwan (China)

    2015-07-01

    Abstract-Multiple-photon emitters, such as In-111 or Se-75, have enormous potential in the field of nuclear medicine imaging. For example, Se-75 can be used to investigate the bile acid malabsorption and measure the bile acid pool loss. The simulation system for emission tomography (SimSET) is a well-known Monte Carlo simulation (MCS) code in nuclear medicine for its high computational efficiency. However, current SimSET cannot simulate these isotopes due to the lack of modeling of complex decay scheme and the time-dependent decay process. To extend the versatility of SimSET for simulation of those multi-photon emission isotopes, a time-resolved multiple photon history generator based on SimSET codes is developed in present study. For developing the time-resolved SimSET (trSimSET) with radionuclide decay process, the new MCS model introduce new features, including decay time information and photon time-of-flight information, into this new code. The half-life of energy states were tabulated from the Evaluated Nuclear Structure Data File (ENSDF) database. The MCS results indicate that the overall percent difference is less than 8.5% for all simulation trials as compared to GATE. To sum up, we demonstrated that time-resolved SimSET multiple photon history generator can have comparable accuracy with GATE and keeping better computational efficiency. The new MCS code is very useful to study the multi-photon imaging of novel isotopes that needs the simulation of lifetime and the time-of-fight measurements. (authors)

  5. Non-invasive in vivo characterization of skin wound healing using label-free multiphoton microscopy (Conference Presentation)

    Science.gov (United States)

    Jones, Jake D.; Majid, Fariah; Ramser, Hallie; Quinn, Kyle P.

    2017-02-01

    Non-healing ulcerative wounds, such as diabetic foot ulcers, are challenging to diagnose and treat due to their numerous possible etiologies and the variable efficacy of advanced wound care products. Thus, there is a critical need to develop new quantitative biomarkers and diagnostic technologies that are sensitive to wound status in order to guide care. The objective of this study was to evaluate the utility of label-free multiphoton microscopy for characterizing wound healing dynamics in vivo and identifying potential differences in diabetic wounds. We isolated and measured an optical redox ratio of FAD/(NADH+FAD) autofluorescence to provide three-dimensional maps of local cellular metabolism. Using a mouse model of wound healing, in vivo imaging at the wound edge identified a significant decrease in the optical redox ratio of the epidermis (p≤0.0103) between Days 3 through 14 compared to Day 1. This decrease in redox ratio coincided with a decrease in NADH fluorescence lifetime and thickening of the epithelium, collectively suggesting a sensitivity to keratinocyte hyperproliferation. In contrast to normal wounds, we have found that keratinocytes from diabetic wounds remain in a proliferative state at later time points with a lower redox ratio at the wound edge. Microstructural organization and composition was also measured from second harmonic generation imaging of collagen and revealed differences between diabetic and non-diabetic wounds. Our work demonstrates label-free multiphoton microscopy offers potential to provide non-invasive structural and functional biomarkers associated with different stages of skin wound healing, which may be used to detect delayed healing and guide treatment.

  6. Label-free in vivo in situ diagnostic imaging by cellular metabolism quantification with a flexible multiphoton endomicroscope (Conference Presentation)

    Science.gov (United States)

    Leclerc, Pierre; Hage, Charles-Henri; Fabert, Marc; Brevier, Julien; O'Connor, Rodney P.; Bardet-Coste, Sylvia M.; Habert, Rémi; Braud, Flavie; Kudlinski, Alexandre; Louradour, Frederic

    2017-02-01

    Multiphoton microscopy is a cutting edge imaging modality leading to increasing advances in biology and also in the clinical field. To use it at its full potential and at the very heart of clinical practice, there have been several developments of fiber-based multiphoton microendoscopes. The application for those probes is now limited by few major restrictions, such as the difficulty to collect autofluorescence signals from tissues and cells theses being inherently weak (e.g. the ones from intracellular NADH or FAD metabolites). This limitation reduces the usefulness of microendoscopy in general, effectively restraining it to morphological imaging modality requiring staining of the tissues. Our aim is to go beyond this limitation, showing for the first time label-free cellular metabolism monitoring, in vivo in situ in real time. The experimental setup is an upgrade of a recently published one (Ducourthial et.al, Scientific Reports, 2016) where femtosecond pulse fiber delivery is further optimized thank's to a new transmissive-GRISM-based pulse stretcher permitting high energy throughput and wide bandwidth. This device allows fast sequential operation with two different excitation wavelengths for efficient two-photon excited NADH and FAD autofluorescence endoscopic detection (i.e. 860 nm for FAD and 760 nm for NADH), enabling cellular optical redox ratio quantification at 8 frames/s. The obtained results on cell models in vitro and also on animal models in vivo (e.g. neurons of a living mouse) prove that we accurately assess the level of NADH and FAD at subcellular resolution through a 3-meters-long fiber with our miniaturized probe (O.D. =2.2 mm).

  7. Comparison of higher-order multiphoton signal generation and collection at the 1700-nm window based on transmittance measurement of objective lenses.

    Science.gov (United States)

    Wen, Wenhui; Wang, Yuxin; Liu, Hongji; Wang, Kai; Qiu, Ping; Wang, Ke

    2018-01-01

    One benefit of excitation at the 1700-nm window is the more accessible modalities of multiphoton signal generation. It is demonstrated here that the transmittance performance of the objective lens is of vital importance for efficient higher-order multiphoton signal generation and collection excited at the 1700-nm window. Two commonly used objective lenses for multiphoton microscopy (MPM) are characterized and compared, one with regular coating and the other with customized coating for high transmittance at the 1700-nm window. Our results show that, fourth harmonic generation imaging of mouse tail tendon and 5-photon fluorescence of carbon quantum dots using the regular objective lens shows an order of magnitude signal higher than those using the customized objective lens. Besides, the regular objective lens also enables a 3-photon fluorescence imaging depth of >1600 μm in mouse brain in vivo. Our results will provide guidelines for objective lens selection for MPM at the 1700-nm window. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Use of multi-photon laser-scanning microscopy to describe the distribution of xenobiotic chemicals in fish early life stages

    International Nuclear Information System (INIS)

    Hornung, Michael W.; Cook, Philip M.; Flynn, Kevin M.; Lothenbach, Doug B.; Johnson, Rodney D.; Nichols, John W.

    2004-01-01

    To better understand the mechanisms by which persistent bioaccumulative toxicants (PBTs) produce toxicity during fish early life stages (ELS), dose-response relationships need to be understood in relation to the dynamic distribution of chemicals in sensitive tissues. In this study, a multi-photon laser scanning microscope (MPLSM) was used to determine the multi-photon excitation spectra of several polyaromatic hydrocarbons (PAHs) and to describe chemical distribution among tissues during fish ELS. The multi-photon excitation spectra revealed intense fluorescent signal from the model fluorophore, pentamethyl-difluoro-boro-indacene (BODIPY[reg], less signal from benzo[a]pyrene and fluoranthene, and no detectable signal from pyrene. The imaging method was tested by exposing newly fertilized medaka (Oryzias latipes) eggs to BODIPY[reg] or fluoranthene for 6 h, followed by transfer to clean media. Embryos and larvae were then imaged through 5 days post-hatch. The two test chemicals partitioned similarly throughout development and differences in fluorescence intensity among tissues were evident to a depth of several hundred microns. Initially, the most intense signal was observed in the oil droplet within the yolk, while a moderate signal was seen in the portion of the yolk containing the yolk-platelets. As embryonic development progressed, the liver biliary system, gall bladder, and intestinal tract accumulated strong fluorescent signal. After hatch, once the gastrointestinal tract was completely developed, most of the fluorescent signal was cleared. The MPLSM is a useful tool to describe the tissue distribution of fluorescent PBTs during fish ELS

  9. Controlling the optical bistability beyond the multi-photon resonance condition in a three-level closed-loop atomic system

    International Nuclear Information System (INIS)

    Mahmoudi, Mohammad; Nozari, Narges; Vafafard, Azar; Sahrai, Mostafa

    2012-01-01

    We investigate the optical bistability behavior of a three-level closed-loop atomic system beyond the multi-photon resonance condition. Using the Floquet decomposition, we solve the time-dependent equations of motion, beyond the multi-photon resonance condition. By identifying the different scattering processes contributing to the medium response, it is shown that in general the optical bistability behavior of the system is not phase-dependent. The phase dependence is due to the scattering of the driving and coupling fields into the probe field at a frequency, which, in general, differs from the probe field frequency. - Highlights: → We investigate optical bistability of a three-level closed-loop atomic system, beyond the multi-photon resonance condition. → By applying Floquet decomposition to the equation of motion, the different scattering processes contributing to the medium response are determined. → It is shown that the phase dependence of optical bistability arises from the scattering of the driving and coupling fields into the probe field frequency.

  10. Order of multiphoton excitation of sulfonium photo-acid generators used in photoresists based on SU-8

    Science.gov (United States)

    Williams, Henry E.; Diaz, Carlos; Padilla, Gabriel; Hernandez, Florencio E.; Kuebler, Stephen M.

    2017-06-01

    Multiphoton lithography (MPL), Z-scan spectroscopy, and quantum chemical calculations were employed to investigate the order of multiphoton excitation that occurs when femtosecond laser pulses are used to excite two sulfonium photo-acid generators (PAGs) commonly used in photoresists based on the cross-linkable epoxide SU-8. The mole-fractions of the mono- and bis-sulfonium forms of these PAGs were determined for the commercially available photoresist SU-8 2075 and for the PAGs alone from a separate source. Both were found to contain similar fractions of the mono- and bis-forms, with the mono form present in the majority. Reichert's method was used to determine the solvatochromic strength of the SU-8 matrix, so that results obtained for the PAGs in SU-8 and in solution could be reliably compared. The PAGs were found to exhibit a minimal solvatochromic shift for a series of solvents that span across the solvatochromic strength of SU-8 itself. Sub-micron-sized features were fabricated in SU-8 2075 by MPL using amplified and continuous-wave mode-locked laser pulses. Analysis of the features as a function of average laser power, scan speed, and excitation wavelength shows that the PAGs can be activated by both two- and three-photon absorption (2PA and 3PA). Which activation mode dominates depends principally upon the excitation wavelength because the average laser powers that can be used with the photoresist are limited by practical considerations. The power must be high enough to effect sufficient cross-linking, yet not so high as to exceed the damage threshold of the material. When the laser pulses have a duration on the order of 100 fs, 3PA dominates at wavelengths near 800 nm, whereas 2PA becomes dominant at wavelengths below 700 nm. These findings are corroborated by open-aperture Z-scan measurements and quantum chemical calculations of the cross-sections for 2PA and 3PA as a function of wavelength.

  11. Atomic Energy Commission Act, 2000 (Act 588)

    International Nuclear Information System (INIS)

    2000-01-01

    Act 588 of the Republic of Ghana entitled, Atomic Energy Commission Act, 2000, amends and consolidates the Atomic Energy Commission Act, 204 of 1963 relating to the establishment of the Atomic Energy Commission. Act 588 makes provision for the Ghana Atomic Energy Commission to establish more institutes for the purpose of research in furtherance of its functions and also promote the commercialization of its research and development results. (E.A.A.)

  12. In vivo detection of basal cell carcinoma: comparison of a reflectance confocal microscope and a multiphoton tomograph

    Science.gov (United States)

    Ulrich, Martina; Klemp, Marisa; Darvin, Maxim E.; König, Karsten; Lademann, Jürgen; Meinke, Martina C.

    2013-06-01

    The standard diagnostic procedure for basal cell carcinoma (BCC) is invasive tissue biopsy with time-consuming histological examination. To reduce the number of biopsies, noninvasive optical methods have been developed providing high-resolution skin examination. We present direct comparison of a reflectance confocal microscope (RLSM) and a multiphoton tomograph (MPT) for BCC diagnosis. Both systems are applied to nine patients prior to surgery, and the results are analyzed, including histological results. Both systems prove suitable for detecting typical characteristics of BCC in various stages. The RLSM allows large horizontal overview images to be obtained, enabling the investigator to find the regions of interest quickly, e.g., BCC nests. Elongated cells and palisading structures are easily recognized using both methods. Due to the higher resolution, changes in nucleus diameter or cytoplasm could be visualized with the MPT. Therefore, the nucleus diameter, nucleus/cytoplasm ratio, and cell density are estimated for normal and BCC cells using the MPT. The nucleus of elongated BCC cells is significantly longer than other measured normal skin cells, whereas the cell density and nucleus/cytoplasm ratio of BCC cannot be significantly distinguished from granular cells.

  13. Multi-photon microscopy with a low-cost and highly efficient Cr:LiCAF laser

    Science.gov (United States)

    Sakadić, Sava; Demirbas, Umit; Mempel, Thorsten R.; Moore, Anna; Ruvinskaya, Svetlana; Boas, David A.; Sennaroglu, Alphan; Kartner, Franz X.; Fujimoto, James G.

    2009-01-01

    Multi-photon microscopy (MPM) is a powerful tool for biomedical imaging, enabling molecular contrast and integrated structural and functional imaging on the cellular and subcellular level. However, the cost and complexity of femtosecond laser sources that are required in MPM are significant hurdles to widespread adoption of this important imaging modality. In this work, we describe femtosecond diode pumped Cr:LiCAF laser technology as a low cost alternative to femtosecond Ti:Sapphire lasers for MPM. Using single mode pump diodes which cost only $150 each, a diode pumped Cr:LiCAF laser generates ~70-fs duration, 1.8-nJ pulses at ~800 nm wavelengths, with a repetition rate of 100 MHz and average output power of 180 mW. Representative examples of MPM imaging in neuroscience, immunology, endocrinology and cancer research using Cr:LiCAF laser technology are presented. These studies demonstrate the potential of this laser source for use in a broad range of MPM applications. PMID:19065223

  14. Multiphoton microscopy observations of 3D elastin and collagen fiber microstructure changes during pressurization in aortic media.

    Science.gov (United States)

    Sugita, Shukei; Matsumoto, Takeo

    2017-06-01

    Elastin and collagen fibers play important roles in the mechanical properties of aortic media. Because knowledge of local fiber structures is required for detailed analysis of blood vessel wall mechanics, we investigated 3D microstructures of elastin and collagen fibers in thoracic aortas and monitored changes during pressurization. Using multiphoton microscopy, autofluorescence images from elastin and second harmonic generation signals from collagen were acquired in media from rabbit thoracic aortas that were stretched biaxially to restore physiological dimensions. Both elastin and collagen fibers were observed in all longitudinal-circumferential plane images, whereas alternate bright and dark layers were observed along the radial direction and were recognized as elastic laminas (ELs) and smooth muscle-rich layers (SMLs), respectively. Elastin and collagen fibers are mainly oriented in the circumferential direction, and waviness of collagen fibers was significantly higher than that of elastin fibers. Collagen fibers were more undulated in longitudinal than in radial direction, whereas undulation of elastin fibers was equibiaxial. Changes in waviness of collagen fibers during pressurization were then evaluated using 2-dimensional fast Fourier transform in mouse aortas, and indices of waviness of collagen fibers decreased with increases in intraluminal pressure. These indices also showed that collagen fibers in SMLs became straight at lower intraluminal pressures than those in EL, indicating that SMLs stretched more than ELs. These results indicate that deformation of the aorta due to pressurization is complicated because of the heterogeneity of tissue layers and differences in elastic properties of ELs, SMLs, and surrounding collagen and elastin.

  15. Modulation of the pupil function of microscope objective lens for multifocal multi-photon microscopy using a spatial light modulator

    Science.gov (United States)

    Matsumoto, Naoya; Okazaki, Shigetoshi; Takamoto, Hisayoshi; Inoue, Takashi; Terakawa, Susumu

    2014-02-01

    We propose a method for high precision modulation of the pupil function of a microscope objective lens to improve the performance of multifocal multi-photon microscopy (MMM). To modulate the pupil function, we adopt a spatial light modulator (SLM) and place it at the conjugate position of the objective lens. The SLM can generate an arbitrary number of spots to excite the multiple fluorescence spots (MFS) at the desired positions and intensities by applying an appropriate computer-generated hologram (CGH). This flexibility allows us to control the MFS according to the photobleaching level of a fluorescent protein and phototoxicity of a specimen. However, when a large number of excitation spots are generated, the intensity distribution of the MFS is significantly different from the one originally designed due to misalignment of the optical setup and characteristics of the SLM. As a result, the image of a specimen obtained using laser scanning for the MFS has block noise segments because the SLM could not generate a uniform MFS. To improve the intensity distribution of the MFS, we adaptively redesigned the CGH based on the observed MFS. We experimentally demonstrate an improvement in the uniformity of a 10 × 10 MFS grid using a dye solution. The simplicity of the proposed method will allow it to be applied for calibration of MMM before observing living tissue. After the MMM calibration, we performed laser scanning with two-photon excitation to observe a real specimen without detecting block noise segments.

  16. Resonant multiphoton ionization of caesium atoms by ultra-short laser pulses at 1.06 μm

    International Nuclear Information System (INIS)

    Lompre, L.A.; Mainfray, G.; Manus, C.; Thebault, J.

    1978-01-01

    This paper reports the four-photon ionization of caesium atoms when the laser frequency is tuned through the resonant three-photon transition 6S → 6F. This experiment was performed by using a tunable-wavelength bandwidth-limited subnanosecond laser pulse at 1.06 μm, in the 10 8 -10 9 W.cm -2 laser intensity range. Pulse widths of 1.5 ns, 50 ps, and 15 ps were used. The resonant character of the multiphoton ionization process was observed, even with the shortest pulse of 15 ps. Nevertheless the influence of a temporal effect is demonstrated according to theoretical predictions. The resonance shift ΔE of the 6S → 6F transition energy was found to be linear with the laser intensity I within the range 10 8 -10 9 W.cm -2 . ΔE = αI, with α = 2 cm -1 /GW.cm -2 . This results confirms previous measurements performed with single-mode 35 ns laser pulses and is in very good agreement with calculated resonance shifts

  17. Multiphoton photochemical crosslinking-based fabrication of protein micropatterns with controllable mechanical properties for single cell traction force measurements

    Science.gov (United States)

    Tong, Ming Hui; Huang, Nan; Zhang, Wei; Zhou, Zhuo Long; Ngan, Alfonso Hing Wan; Du, Yanan; Chan, Barbara Pui

    2016-01-01

    Engineering 3D microstructures with predetermined properties is critical for stem cell niche studies. We have developed a multiphoton femtosecond laser-based 3D printing platform, which generates complex protein microstructures in minutes. Here, we used the platform to test a series of fabrication and reagent parameters in precisely controlling the mechanical properties of protein micropillars. Atomic force microscopy was utilized to measure the reduced elastic modulus of the micropillars, and transmission electron microscopy was used to visualize the porosity of the structures. The reduced elastic modulus of the micropillars associated positively and linearly with the scanning power. On the other hand, the porosity and pore size of the micropillars associated inversely and linearly with the scanning power and reagent concentrations. While keeping the elastic modulus constant, the stiffness of the micropillars was controlled by varying their height. Subsequently, the single cell traction forces of rabbit chondrocytes, human dermal fibroblasts, human mesenchymal stem cells, and bovine nucleus pulposus cells (bNPCs) were successfully measured by culturing the cells on micropillar arrays of different stiffness. Our results showed that the traction forces of all groups showed positive relationship with stiffness, and that the chondrocytes and bNPCs generated the highest and lowest traction forces, respectively.

  18. Ex vivo multiscale quantitation of skin biomechanics in wild-type and genetically-modified mice using multiphoton microscopy

    Science.gov (United States)

    Bancelin, Stéphane; Lynch, Barbara; Bonod-Bidaud, Christelle; Ducourthial, Guillaume; Psilodimitrakopoulos, Sotiris; Dokládal, Petr; Allain, Jean-Marc; Schanne-Klein, Marie-Claire; Ruggiero, Florence

    2015-12-01

    Soft connective tissues such as skin, tendon or cornea are made of about 90% of extracellular matrix proteins, fibrillar collagens being the major components. Decreased or aberrant collagen synthesis generally results in defective tissue mechanical properties as the classic form of Elhers-Danlos syndrome (cEDS). This connective tissue disorder is caused by mutations in collagen V genes and is mainly characterized by skin hyperextensibility. To investigate the relationship between the microstructure of normal and diseased skins and their macroscopic mechanical properties, we imaged and quantified the microstructure of dermis of ex vivo murine skin biopsies during uniaxial mechanical assay using multiphoton microscopy. We used two genetically-modified mouse lines for collagen V: a mouse model for cEDS harboring a Col5a2 deletion (a.k.a. pN allele) and the transgenic K14-COL5A1 mice which overexpress the human COL5A1 gene in skin. We showed that in normal skin, the collagen fibers continuously align with stretch, generating the observed increase in mechanical stress. Moreover, dermis from both transgenic lines exhibited altered collagen reorganization upon traction, which could be linked to microstructural modifications. These findings show that our multiscale approach provides new crucial information on the biomechanics of dermis that can be extended to all collagen-rich soft tissues.

  19. Multiphoton spectral analysis of benzo[a]pyrene uptake and metabolism in a rat liver cell line

    International Nuclear Information System (INIS)

    Barhoumi, Rola; Mouneimne, Youssef; Ramos, Ernesto; Morisseau, Christophe; Hammock, Bruce D.; Safe, Stephen; Parrish, Alan R.; Burghardt, Robert C.

    2011-01-01

    Dynamic analysis of the uptake and metabolism of polycyclic aromatic hydrocarbons (PAHs) and their metabolites within live cells in real time has the potential to provide novel insights into genotoxic and non-genotoxic mechanisms of cellular injury caused by PAHs. The present work, combining the use of metabolite spectra generated from metabolite standards using multiphoton spectral analysis and an 'advanced unmixing process', identifies and quantifies the uptake, partitioning, and metabolite formation of one of the most important PAHs (benzo[a]pyrene, BaP) in viable cultured rat liver cells over a period of 24 h. The application of the advanced unmixing process resulted in the simultaneous identification of 8 metabolites in live cells at any single time. The accuracy of this unmixing process was verified using specific microsomal epoxide hydrolase inhibitors, glucuronidation and sulfation inhibitors as well as several mixtures of metabolite standards. Our findings prove that the two-photon microscopy imaging surpasses the conventional fluorescence imaging techniques and the unmixing process is a mathematical technique that seems applicable to the analysis of BaP metabolites in living cells especially for analysis of changes of the ultimate carcinogen benzo[a]pyrene-r-7,t-8-dihydrodiol-t-9,10-epoxide. Therefore, the combination of the two-photon acquisition with the unmixing process should provide important insights into the cellular and molecular mechanisms by which BaP and other PAHs alter cellular homeostasis.

  20. Real-time visualization of melanin granules in normal human skin using combined multiphoton and reflectance confocal microscopy.

    Science.gov (United States)

    Majdzadeh, Ali; Lee, Anthony M D; Wang, Hequn; Lui, Harvey; McLean, David I; Crawford, Richard I; Zloty, David; Zeng, Haishan

    2015-05-01

    Recent advances in biomedical optics have enabled dermal and epidermal components to be visualized at subcellular resolution and assessed noninvasively. Multiphoton microscopy (MPM) and reflectance confocal microscopy (RCM) are noninvasive imaging modalities that have demonstrated promising results in imaging skin micromorphology, and which provide complementary information regarding skin components. This study assesses whether combined MPM/RCM can visualize intracellular and extracellular melanin granules in the epidermis and dermis of normal human skin. We perform MPM and RCM imaging of in vivo and ex vivo skin in the infrared domain. The inherent three-dimensional optical sectioning capability of MPM/RCM is used to image high-contrast granular features across skin depths ranging from 50 to 90 μm. The optical images thus obtained were correlated with conventional histologic examination including melanin-specific staining of ex vivo specimens. MPM revealed highly fluorescent granular structures below the dermal-epidermal junction (DEJ) region. Histochemical staining also demonstrated melanin-containing granules that correlate well in size and location with the granular fluorescent structures observed in MPM. Furthermore, the MPM fluorescence excitation wavelength and RCM reflectance of cell culture-derived melanin were equivalent to those of the granules. This study suggests that MPM can noninvasively visualize and quantify subepidermal melanin in situ. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  1. Identification of the boundary between normal breast tissue and invasive ductal carcinoma during breast-conserving surgery using multiphoton microscopy

    Science.gov (United States)

    Deng, Tongxin; Nie, Yuting; Lian, Yuane; Wu, Yan; Fu, Fangmeng; Wang, Chuan; Zhuo, Shuangmu; Chen, Jianxin

    2014-11-01

    Breast-conserving surgery has become an important way of surgical treatment for breast cancer worldwide nowadays. Multiphoton microscopy (MPM) has the ability to noninvasively visualize tissue architectures at the cellular level using intrinsic fluorescent molecules in biological tissues without the need for fluorescent dye. In this study, MPM is used to image the microstructures of terminal duct lobular unit (TDLU), invasive ductal carcinoma and the boundary region between normal and cancerous breast tissues. Our study demonstrates that MPM has the ability to not only reveal the morphological changes of the cuboidal epithelium, basement membrane and interlobular stroma but also identify the boundary between normal breast tissue and invasive ductal carcinoma, which correspond well to the Hematoxylin and Eosin (H and E) images. Predictably, MPM can monitor surgical margins in real time and provide considerable accuracy for resection of breast cancerous tissues intraoperatively. With the development of miniature, real-time MPM imaging technology, MPM should have great application prospects during breast-conserving surgery.

  2. O2 rotational temperature measurements in an atmospheric air microdischarge by radar resonance-enhanced multiphoton ionization

    International Nuclear Information System (INIS)

    Sawyer, Jordan; Wu, Yue; Zhang, Zhili; Adams, Steven F.

    2013-01-01

    Nonintrusive spatially resolved rotational temperature measurements in an atmospheric air microdischarge are presented. The measurements were based on coherent microwave Rayleigh scattering (Radar) from resonance-enhanced multiphoton ionization of molecular oxygen. The open air DC microdischarge source operated in a stable “normal-glow” mode and pin-to-pin electrodes spaced 1.3 mm apart. The second harmonic of a tunable dye laser beam was focused between the two electrodes and scanned between 286 and 288 nm. Coherent microwave Rayleigh scattering was used to collect the two-photon rotational spectra of O 2 at C 3 Π(v = 2)←X 3 Σ(v′ = 0) transitions. The Boltzmann plots from analyses of the O 2 rotational lines determined local rotational temperatures at various axial locations between the electrodes. The molecular oxygen rotational temperature varied from ∼1150 K to ∼1350 K within the discharge area. The measurements had an accuracy of ∼±50 K.

  3. O2 rotational temperature measurements in an atmospheric air microdischarge by radar resonance-enhanced multiphoton ionization

    Science.gov (United States)

    Sawyer, Jordan; Wu, Yue; Zhang, Zhili; Adams, Steven F.

    2013-06-01

    Nonintrusive spatially resolved rotational temperature measurements in an atmospheric air microdischarge are presented. The measurements were based on coherent microwave Rayleigh scattering (Radar) from resonance-enhanced multiphoton ionization of molecular oxygen. The open air DC microdischarge source operated in a stable "normal-glow" mode and pin-to-pin electrodes spaced 1.3 mm apart. The second harmonic of a tunable dye laser beam was focused between the two electrodes and scanned between 286 and 288 nm. Coherent microwave Rayleigh scattering was used to collect the two-photon rotational spectra of O2 at C3Π(v = 2)←X3Σ(v' = 0) transitions. The Boltzmann plots from analyses of the O2 rotational lines determined local rotational temperatures at various axial locations between the electrodes. The molecular oxygen rotational temperature varied from ˜1150 K to ˜1350 K within the discharge area. The measurements had an accuracy of ˜±50 K.

  4. Delivery of cyclodextrin polymers to bacterial biofilms - An exploratory study using rhodamine labelled cyclodextrins and multiphoton microscopy.

    Science.gov (United States)

    Thomsen, Hanna; Benkovics, Gábor; Fenyvesi, Éva; Farewell, Anne; Malanga, Milo; Ericson, Marica B

    2017-10-15

    Cyclodextrin (CD) polymers are interesting nanoparticulate systems for pharmaceutical delivery; however, knowledge regarding their applications towards delivery into complex microbial biofilm structures is so far limited. The challenge is to demonstrate penetration and transport through the biofilm and its exopolysaccharide matrix. The ideal functionalization for penetration into mature biofilms is unexplored. In this paper, we present a novel set of rhodamine labelled βCD-polymers, with different charge moieties, i.e., neutral, anionic, and cationic, and explore their potential delivery into mature Staphylococcus epidermidis biofilms using multiphoton laser scanning microscopy (MPM). The S. epidermidis biofilms, being a medically relevant model organism, were stained with SYTO9. By using MPM, three-dimensional imaging and spectral investigation of the distribution of the βCD-polymers could be obtained. It was found that the cationic βCD-polymers showed significantly higher integration into the biofilms, compared to neutral and anionic functionalized βCDs. None of the carriers presented any inherent toxicity to the biofilms, meaning that the addition of rhodamine moiety does not affect the inertness of the delivery system. Taken together, this study demonstrates a novel approach by which delivery of fluorescently labelled CD nanoparticles to bacterial biofilms can be explored using MPM. Future studies should be undertaken investigating the potential in using cationic functionalization of CD based delivery systems for targeting anti-microbial effects in biofilms. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  5. Nuclear Regulatory Authority Act, 2015 (Act 895)

    International Nuclear Information System (INIS)

    2015-04-01

    An Act to establish a Nuclear Regulatory Authority in Ghana. This Act provides for the regulation and management of activities and practices for the peaceful use of nuclear material or energy, and to provide for the protection of persons and the environment against the harmful effects of radiation; and to ensure the effective implementation of the country’s international obligations and for related matters. This Act replaced the Radiation Protection Instrument, of 1993 (LI 1559).

  6. Visualizing Angiogenesis by Multiphoton Microscopy In Vivo in Genetically Modified 3D-PLGA/nHAp Scaffold for Calvarial Critical Bone Defect Repair.

    Science.gov (United States)

    Li, Jian; Jahr, Holger; Zheng, Wei; Ren, Pei-Gen

    2017-09-07

    The reconstruction of critically sized bone defects remains a serious clinical problem because of poor angiogenesis within tissue-engineered scaffolds during repair, which gives rise to a lack of sufficient blood supply and causes necrosis of the new tissues. Rapid vascularization is a vital prerequisite for new tissue survival and integration with existing host tissue. The de novo generation of vasculature in scaffolds is one of the most important steps in making bone regeneration more efficient, allowing repairing tissue to grow into a scaffold. To tackle this problem, the genetic modification of a biomaterial scaffold is used to accelerate angiogenesis and osteogenesis. However, visualizing and tracking in vivo blood vessel formation in real-time and in three-dimensional (3D) scaffolds or new bone tissue is still an obstacle for bone tissue engineering. Multiphoton microscopy (MPM) is a novel bio-imaging modality that can acquire volumetric data from biological structures in a high-resolution and minimally-invasive manner. The objective of this study was to visualize angiogenesis with multiphoton microscopy in vivo in a genetically modified 3D-PLGA/nHAp scaffold for calvarial critical bone defect repair. PLGA/nHAp scaffolds were functionalized for the sustained delivery of a growth factor pdgf-b gene carrying lentiviral vectors (LV-pdgfb) in order to facilitate angiogenesis and to enhance bone regeneration. In a scaffold-implanted calvarial critical bone defect mouse model, the blood vessel areas (BVAs) in PHp scaffolds were significantly higher than in PH scaffolds. Additionally, the expression of pdgf-b and angiogenesis-related genes, vWF and VEGFR2, increased correspondingly. MicroCT analysis indicated that the new bone formation in the PHp group dramatically improved compared to the other groups. To our knowledge, this is the first time multiphoton microscopy was used in bone tissue-engineering to investigate angiogenesis in a 3D bio-degradable scaffold in

  7. Comparative study of Nd(3+) emission from 4f2 5d and 4f3 configurations induced by multiphotonic process in YLF, GLF and LLF crystals

    International Nuclear Information System (INIS)

    Librantz, Andre Felipe Henriques

    2000-01-01

    Nd 3+ ultraviolet fluorescence induced by multiphotonic laser excitations was studied in Nd-doped YLiF 4 (YLF) and LuLiF 4 (LLF) crystals by using the time resolved spectroscopy technique. The UV luminescences are due to transitions between the 4f 2 5d and the 4f 3 electronic configurations of Nd 3+ ions. The 4f 2 5d configuration can be reached by direct pumping the UV transition or by multiphotonic excitation, both processes give raise to the UV emission band with a structure due to the strong phonon coupling, expected for a 5d orbital involvement in the transition. The multiphotonic excitation process is due to three photons (532 nm) sequential absorptions of 532 nm-photons by metastable levels of the 4f 3 configuration splitted by crystalline local field. The sequential excitation of Nd by the pumping laser is attributed to the 4 I 9/2 +532nm → 4 G 7/2 ground state absorption followed by the 4 G 7/2 +532 nm →2 F 5/2 and 2 F 5/2 +532 nm → 4f 2 5d excited state absorptions. The UV emissions due to 4f 2 5d configuration are parity allowed, having lifetime of 35 ns in contrast to UV emissions from 4f 3 configuration which are induced by two absorption steps and are parity forbidden showing longer lifetime of 8μs and narrow tines. The polarization effects of the UV emissions were studied and their behavior are dependent on the excited state configuration involving or not the 5d orbital. The allowed UV emissions positions were affected by the host variation more than the ones originating from the 4f 3 configuration as expected. The electronic energy of the 4f 2 5d configuration shifts to lower energy when increasing the crystal field. (author)

  8. Infrared multiphoton dissociation of acrolein. Time-resolved observation of CO ( v = 1) IR emission at 4.7 μm

    Science.gov (United States)

    Chowdhury, P. K.; Rama Rao, K. V. S.; Mittal, J. P.

    1994-02-01

    In contrast to the photochemistry of electronically excited acrolein producing vinyl and formyl radicals via CC bond rupture, multiphoton vibrationally excited molecules undergo concerted dissociation generating CO and ethylene. Vibrational excitation in the CO product is detected immediately following the CO 2 laser pulse by observing IR emission at 4.7 μm. The decay of the IR emission was studied as a function of acrolein pressure. A vibrational-vibrational relaxation rate constant of CO ( v=1) by acrolein is found to be 1240 ± 200 Torr -1 s -1.

  9. Clinical coherent anti-Stokes Raman scattering and multiphoton tomography of human skin with a femtosecond laser and photonic crystal fiber

    International Nuclear Information System (INIS)

    Breunig, Hans Georg; Weinigel, Martin; Bückle, Rainer; Kellner-Höfer, Marcel; König, Karsten; Lademann, Jürgen; Darvin, Maxim E; Sterry, Wolfram

    2013-01-01

    We report on in vivo coherent anti-Stokes Raman scattering spectroscopy (CARS), two-photon fluorescence and second-harmonic-generation imaging on human skin with a novel multimodal clinical CARS/multiphoton tomograph. CARS imaging is realized by a combination of femtosecond pulses with broadband continuum pulses generated by a photonic crystal fiber. The images reveal the microscopic distribution of (i) non-fluorescent lipids, (ii) endogenous fluorophores and (iii) the collagen network inside the human skin in vivo with subcellular resolution. Examples of healthy as well as cancer-affected skin are presented. (letter)

  10. Signatures of tunneling and multiphoton ionization in the electron-momentum distributions of atoms by intense few-cycle laser pulses

    International Nuclear Information System (INIS)

    Wickenhauser, M.; Tong, X. M.; Arbo, D. G.; Burgdoerfer, J.; Lin, C. D.

    2006-01-01

    Electron-momentum distributions for above-threshold ionization of argon in a few-cycle, linearly polarized laser pulse are investigated. Spectral features characteristic of multiphoton as well as tunneling ionization coexist over a range of the Keldysh parameter γ in the transition regime γ∼1. Surprisingly, the simple strong-field approximation (SFA) is capable of reproducing the key features of the two-dimensional momentum distributions found in the full solution of the time-dependent Schroedinger equation, despite the fact that SFA is known to severely underestimate the total ionization probability

  11. Clinical coherent anti-Stokes Raman scattering and multiphoton tomography of human skin with a femtosecond laser and photonic crystal fiber

    Science.gov (United States)

    Breunig, Hans Georg; Weinigel, Martin; Bückle, Rainer; Kellner-Höfer, Marcel; Lademann, Jürgen; Darvin, Maxim E.; Sterry, Wolfram; König, Karsten

    2013-02-01

    We report on in vivo coherent anti-Stokes Raman scattering spectroscopy (CARS), two-photon fluorescence and second-harmonic-generation imaging on human skin with a novel multimodal clinical CARS/multiphoton tomograph. CARS imaging is realized by a combination of femtosecond pulses with broadband continuum pulses generated by a photonic crystal fiber. The images reveal the microscopic distribution of (i) non-fluorescent lipids, (ii) endogenous fluorophores and (iii) the collagen network inside the human skin in vivo with subcellular resolution. Examples of healthy as well as cancer-affected skin are presented.

  12. Multiphoton ionization of many-electron atoms and highly-charged ions in intense laser fields: a relativistic time-dependent density functional theory approach

    Science.gov (United States)

    Tumakov, Dmitry A.; Telnov, Dmitry A.; Maltsev, Ilia A.; Plunien, Günter; Shabaev, Vladimir M.

    2017-10-01

    We develop an efficient numerical implementation of the relativistic time-dependent density functional theory (RTDDFT) to study multielectron highly-charged ions subject to intense linearly-polarized laser fields. The interaction with the electromagnetic field is described within the electric dipole approximation. The resulting time-dependent relativistic Kohn-Sham (RKS) equations possess an axial symmetry and are solved accurately and efficiently with the help of the time-dependent generalized pseudospectral method. As a case study, we calculate multiphoton ionization probabilities of the neutral argon atom and argon-like xenon ion. Relativistic effects are assessed by comparison of our present results with existing non-relativistic data.

  13. Nanostructures based on quantum dots for application in promising methods of single- and multiphoton imaging and diagnostics

    Science.gov (United States)

    Nabiev, I. R.

    2017-01-01

    Molecules recognizing biomarkers of diseases (monoclonal antibodies (monoABs)) are often too large for biomedical applications, and the conditions that are used to bind them with nanolabels lead to disordered orientation of monoABs with respect to the nanoparticle surface. Extremely small nanoprobes, designed via oriented conjugation of quantum dots (QDs) with single-domain antibodies (sdABs) derived from the immunoglobulin of llama and produced in the E. coli culture, have a hydrodynamic diameter less than 12 nm and contain equally oriented sdAB molecules on the surface of each QD. These nanoprobes exhibit excellent specificity and sensitivity in quantitative determination of a small number of cells expressing biomarkers. In addition, the higher diffusion coefficient of sdABs makes it possible to perform immunohistochemical analysis in bulk tissue, inaccessible for conventional monoABs. The necessary conditions for implementing high-quality immunofluorescence diagnostics are a high specificity of labeling and clear differences between the fluorescence of nanoprobes and the autofluorescence of tissues. Multiphoton micros-copy with excitation in the near-IR spectral range, which is remote from the range of tissue autofluorescence excitation, makes it possible to solve this problem and image deep layers in biological tissues. The two-photon absorption cross sections of CdSe/ZnS QDs conjugated with sdABs exceed the corresponding values for organic fluorophores by several orders of magnitude. These nanoprobes provide clear discrimination between the regions of tumor and normal tissues with a ratio of the sdAB fluorescence to the tissue autofluorescence upon two-photon excitation exceeding that in the case of single-photon excitation by a factor of more than 40. The data obtained indicate that the sdAB-QD conjugates used as labels provide the same, or even better, quality as the "gold standard" of immunohistochemical diagnostics. The developed nanoprobes are expected to

  14. A new method using multiphoton imaging and morphometric analysis for differentiating chromophobe renal cell carcinoma and oncocytoma kidney tumors

    Science.gov (United States)

    Wu, Binlin; Mukherjee, Sushmita; Jain, Manu

    2016-03-01

    Distinguishing chromophobe renal cell carcinoma (chRCC) from oncocytoma on hematoxylin and eosin images may be difficult and require time-consuming ancillary procedures. Multiphoton microscopy (MPM), an optical imaging modality, was used to rapidly generate sub-cellular histological resolution images from formalin-fixed unstained tissue sections from chRCC and oncocytoma.Tissues were excited using 780nm wavelength and emission signals (including second harmonic generation and autofluorescence) were collected in different channels between 390 nm and 650 nm. Granular structure in the cell cytoplasm was observed in both chRCC and oncocytoma. Quantitative morphometric analysis was conducted to distinguish chRCC and oncocytoma. To perform the analysis, cytoplasm and granules in tumor cells were segmented from the images. Their area and fluorescence intensity were found in different channels. Multiple features were measured to quantify the morphological and fluorescence properties. Linear support vector machine (SVM) was used for classification. Re-substitution validation, cross validation and receiver operating characteristic (ROC) curve were implemented to evaluate the efficacy of the SVM classifier. A wrapper feature algorithm was used to select the optimal features which provided the best predictive performance in separating the two tissue types (classes). Statistical measures such as sensitivity, specificity, accuracy and area under curve (AUC) of ROC were calculated to evaluate the efficacy of the classification. Over 80% accuracy was achieved as the predictive performance. This method, if validated on a larger and more diverse sample set, may serve as an automated rapid diagnostic tool to differentiate between chRCC and oncocytoma. An advantage of such automated methods are that they are free from investigator bias and variability.

  15. Examination of diagnostic features in multiphoton microscopy and optical coherence tomography images of ovarian tumorigenesis in a mouse model

    Science.gov (United States)

    Watson, Jennifer M.

    Ovarian cancer is a deadly disease owing to the non-specific symptoms and suspected rapid progression, leading to frequent late stage detection and poor prognosis. Medical imaging methods such as CT, MRI and ultrasound as well as serum testing for cancer markers have had extremely poor performance for early disease detection. Due to the poor performance of available screening methods, and the impracticality and ineffectiveness of taking tissue biopsies from the ovary, women at high risk for developing ovarian cancer are often advised to undergo prophylactic salpingo-oophorectomy. This surgery results in many side effects and is most often unnecessary since only a fraction of high risk women go on to develop ovarian cancer. Better understanding of the early development of ovarian cancer and characterization of morphological changes associated with early disease could lead to the development of an effective screening test for women at high risk. Optical imaging methods including optical coherence tomography (OCT) and multiphoton microscopy (MPM) are excellent tools for studying disease progression owing to the high resolution and depth sectioning capabilities. Further, these techniques are excellent for optical biopsy because they can image in situ non-destructively. In the studies described in this dissertation OCT and MPM are used to identify cellular and tissue morphological changes associated with early tumor development in a mouse model of ovarian cancer. This work is organized into three specific aims. The first aim is to use the images from the MPM phenomenon of second harmonic generation to quantitatively examine the morphological differences in collagen structure in normal mouse ovarian tissue and mouse ovarian tumors. The second aim is to examine the differences in endogenous two-photon excited fluorescence in normal mouse ovarian tissue and mouse ovarian tumors. The third and final aim is to identify changes in ovarian microstructure resulting from early

  16. Energy distributions of neutral species ejected from well-characterized surfaces measured by means of multiphoton resonance ionization spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Ishikawa, D.; Ishigami, R.; Dhole, S.D.; Morita, K. E-mail: k-morita@mail.nucl.nagoya-u.ac.jp

    2000-04-01

    The energy distributions of neutral atoms ejected from the polycrystalline Cu target, the Si(1 1 1)-7x7 surface, and the Si(1 1 1)-''5 x 5''-Cu surface by 5 keV Ar{sup +} ion bombardment have been measured with very high efficiency by means of the multi-photon resonance ionization spectroscopy, in order to obtain the surface binding energies. The energy distributions for Cu from polycrystalline Cu target, Si from the Si(1 1 1)-7x7 surface, and Cu from the Si(1 1 1)-''5 x 5''-Cu surface have been found to have a peak at energies of around 3.0, 5.0 and 1.5 eV, and the function shapes of high energy tails to be proportional to E{sup -1.9}, E{sup -1.2} and E{sup -1.3}, respectively. Based on the linear collision cascade theory, the surface binding energies are determined to be 5.7, 6.0 and 2.0 eV, and the power factor m in the power law approximation to the Thomas-Fermi potential are determined to be 0.1, 0.4 and 0.3 for Cu from the Cu polycrystalline, Si from the Si(1 1 1)-7x7 surface, and Cu from the Si(1 1 1)-''5 x 5''-Cu surface, respectively. In conclusion, the results indicate that the energy distributions of ejected particles are well characterized by the linear collision cascade theory developed by Sigmund.

  17. Modeling extracellular matrix (ECM) alterations in ovarian cancer by multiphoton excited fabrication of stromal models (Conference Presentation)

    Science.gov (United States)

    Campagnola, Paul J.; Ajeti, Visar; Lara, Jorge; Eliceiri, Kevin W.; Patankar, Mansh

    2016-04-01

    A profound remodeling of the extracellular matrix (ECM) occurs in human ovarian cancer but it unknown how this affects tumor growth, where this understanding could lead to better diagnostics and therapeutic approaches. We investigate the role of these ECM alterations by using multiphoton excited (MPE) polymerization to fabricate biomimetic models to investigate operative cell-matrix interactions in invasion/metastasis. First, we create nano/microstructured gradients mimicking the basal lamina to study adhesion/migration dynamics of ovarian cancer cells of differing metastatic potential. We find a strong haptotactic response that depends on both contact guidance and ECM binding cues. While we found enhanced migration for more invasive cells, the specifics of alignment and directed migration also depend on cell polarity. We further use MPE fabrication to create collagen scaffolds with complex, 3D submicron morphology. The stromal scaffold designs are derived directly from "blueprints" based on SHG images of normal, high risk, and malignant ovarian tissues. The models are seeded with different cancer cell lines and this allows decoupling of the roles of cell characteristics (metastatic potential) and ECM structure and composition (normal vs cancer) on adhesion/migration dynamics. We found the malignant stroma structure promotes enhanced migration and proliferation and also cytoskeletal alignment. Creating synthetic models based on fibers patterns further allows decoupling the topographic roles of the fibers themselves vs their alignment within the tissue. These models cannot be synthesized by other conventional fabrication methods and we suggest the MPE image-based fabrication method will enable a variety of studies in cancer biology.

  18. Theory of the effect of odd-photon destructive interference on optical shifts in resonantly enhanced multiphoton excitation and ionization

    International Nuclear Information System (INIS)

    Payne, M.G.; Deng, L.; Garrett, W.R.

    1998-01-01

    We present a theory for two- and three-photon excitation, optical shifting, and four-wave mixing when a first laser is tuned onto, or near, a two-photon resonance and a second much more intense laser is tuned near or on resonance between the two-photon resonance and a second excited state. When the second excited state has a dipole-allowed transition back to the ground state and the concentration is sufficiently high, a destructive interference is produced between three-photon coupling of the ground state and the second excited state and one-photon coupling between the same states by the internally generated four-wave mixing field. This interference leads to several striking effects. For instance, as the onset of the interference occurs, the optical shifts in the two-photon resonance excitation line shape become smaller in copropagating geometry so that the line shapes for multiphoton ionization enhanced by the two-photon resonance eventually become unaffected by the second laser. In the same range of concentrations the four-wave mixing field evolves to a concentration-independent intensity. With counterpropagating laser beams the line shape exhibits normal optical shifts like those observed for both copropagating and counterpropagating laser beams at very low concentrations. The theoretical work presented here extends our earlier works by including the effect of laser bandwidth and by removing the restriction of having the second laser be tuned far from three-photon resonance. In this way we have now included, as a special case, the effect of both laser bandwidth and interference on laser-induced transparency. Unlike other effects related to odd-photon destructive interference, the effect of a broad bandwidth is to bring about the predicted effects at much lower concentrations. Studies in rubidium show good agreement between theory and experiment for both ionization line shapes and four-wave mixing intensity as a function of concentration. copyright 1998 The

  19. Fast monitoring of motor exhaust components by resonant multi-photon ionisation and time-of-flight mass spectrometry

    Science.gov (United States)

    Franzen, Jochen; Frey, Rüdiger; Nagel, Holger

    1995-03-01

    A new analytical procedure is provided by the combination of two types of spectroscopy. Resonant ionization of selected compounds by multiphoton ionization is based on results of absorption spectroscopy for the compound molecules of interest and time-of-flight mass spectrometry serves for the unambigious detection of these compounds. An interesting application of this method is the fast exhaust gas analysis. In the development of future combustion engines, the management of dynamic motor processes becomes predominant because by more than 90 % of all the dangerous exhaust pollutions are produced in instationary motor phases such as fast speed or load changes. The investigation of dynamic processes however, requires fast analytical procedures with millisecond time resolution together with the capability to measure individual components in a very complex gas mixture The objectives for a development project of such an instrument were set by the Research Association for Combustion Engines (Forschungsvereinigung Verbrennungskraftmaschinen, FVV, Germany): Up to ten substances should be monitored synchroneously with a time resolution of about 10 milliseconds, with concentration limits of 1 part per million and with a precision better than 10 % relative standard deviation. Such a laser mass spectrometer for fast multi-component automotive exhaust analyses has been developed in a joint research project by Bruker-Franzen Analytik GmbH, Dornier GmbH and the Technical University of Munich. The system has been applied at a motor test facility to investigate the emissions of the aromatic hydrocarbons benzene, toluene and xylene, of nitric oxide and acetaldehyde in stationary and dynamic engine operation. These measurements demonstrate that strong emission of these pollutants takes place at instationary engine operation and in particular that these compounds are emitted at different times, giving new information about the processes in the combustion chamber and in the exhaust pipe.

  20. Atomic Act amended

    International Nuclear Information System (INIS)

    Drabova, D.

    2002-01-01

    In the paper by the chairwoman of the Czech nuclear regulatory authority, the history of Czech nuclear legislation is outlined, the reasons for the amendment of the Atomic Act (Act No. 18/1997) are explained, and the amendments themselves are highlighted. The Act No. 13/2002 of 18 December 2001 is reproduced from the official Collection of Acts of the Czech Republic in the facsimile form. The following acts were thereby amended: Atomic Act No. 18/1997, Metrology Act No. 505/1990, Public Health Protection Act No. 258/2000, and Act No. 2/1969 on the Establishment of Ministries and Other Governmental Agencies of the Czech Republic. (P.A.)

  1. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... Therapeutics Lab Developing New Treatments The CF Foundation offers a number of resources for learning about clinical ... her father: Demonstrate and discuss common ACT therapies Offer their tips for fitting ACTs into daily life ...

  2. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... Treatments and Therapies Airway Clearance Airway Clearance Techniques (ACTs) There are different ways to clear your airways. ... or caregiver. Older kids and adults can choose ACTs that they can do on their own. Share ...

  3. Nuclear Installations Act 1965

    International Nuclear Information System (INIS)

    1965-01-01

    This Act governs all activities related to nuclear installations in the United Kingdom. It provides for the licensing procedure for nuclear installations, the duties of licensees, the competent authorities and carriers of nuclear material in respect of nuclear occurrences, as well as for the system of third party liability and compensation for nuclear damage. The Act repeals the Nuclear Installations (Licensing and Insurance) Act 1959 and the Nuclear Installations (Amendment Act) 1965 except for its Section 17(2). (NEA) [fr

  4. Energetics of the rearrangement of neutral and ionized perfluorocyclopropane to perfluoropropylene. Use of infrared multiphoton dissociation spectra to identify structural isomers of molecular ions

    International Nuclear Information System (INIS)

    Bomse, D.S.; Berman, D.W.; Beauchamp, J.L.

    1981-01-01

    Infrared photodissociation spectroscopy is used to compare the structure of gas-phase C 3 F 6 + ions obtained by electron-impact ionization of two isomeric precursors: perfluoropropylene and perfluorocyclopropane. Photodissociation spectra are obtained by observing the extent of multiphoton dissociation as the CO 2 laser is tuned across the 925 to 1080 cm -1 wavelength range. Ions are formed, stored, and detected with the use of techniques of ion cyclotron resonance spectroscopy. Infrared multiphoton excitation is effected by using low-power, continuous-wave laser radiation. The fingerprint spectrum of the molecular ion of perfluorocyclopropane is identical with that obtained from perfluoropropylene, indicating rearrangement of the former to the latter. Photodissociation kinetics indicate that the entire perfluorocyclopropane molecular ion population isomerizes to the more stable perfluoropropylene structure. Thermochemistry of C 3 F 6 and C 3 F 6 + isomers is discussed. Comparisons are made with the analogous C 3 H 6 system. Photoionization mass spectroscopy results yield ΔH/sub f/(c-C 3 F 6 ) = -233.8 kcal/mol. 4 figures

  5. Multi-photon vertical cross-sectional imaging with a dynamically-balanced thin-film PZT z-axis microactuator.

    Science.gov (United States)

    Choi, Jongsoo; Duan, Xiyu; Li, Haijun; Wang, Thomas D; Oldham, Kenn R

    2017-10-01

    Use of a thin-film piezoelectric microactuator for axial scanning during multi-photon vertical cross-sectional imaging is described. The actuator uses thin-film lead-zirconate-titanate (PZT) to generate upward displacement of a central mirror platform, micro-machined from a silicon-on-insulator (SOI) wafer to dimensions compatible with endoscopic imaging instruments. Device modeling in this paper focuses on existence of frequencies near device resonance producing vertical motion with minimal off-axis tilt even in the presence of multiple vibration modes and non-uniformity in fabrication outcomes. Operation near rear resonance permits large stroke lengths at low voltages relative to other vertical microactuators. Highly uniform vertical motion of the mirror platform is a key requirement for vertical cross-sectional imaging in the remote scan architecture being used for multi-photon instrument prototyping. The stage is installed in a benchtop testbed in combination with an electrostatic mirror that performs in-plane scanning. Vertical sectional images are acquired from 15 μm diameter beads and excised mouse colon tissue.

  6. Evaluation of Elastin/Collagen Content in Human Dermis in-Vivo by Multiphoton Tomography—Variation with Depth and Correlation with Aging

    Directory of Open Access Journals (Sweden)

    Jean-Christophe Pittet

    2014-08-01

    Full Text Available The aim of this study was to evaluate the influence of the depth of the dermis on the measured collagen and elastin levels and to establish the correlation between the amount of these two extracellular matrix (ECM components and age. Multiphoton Microscopy (MPM that measures the autofluorescence (AF and second harmonic generation (SHG was used to quantify the levels of elastin and collagen and to determine the SAAID (SHG-to-AF Aging Index of Dermis at two different skin depths. A 50 MHz ultrasound scanner was used for the calculation of the Sub Epidermal Non Echogenic Band (SENEB. The measurements of the skin mechanical properties were done with a cutometer. All measurements were performed on two groups of 30 healthy female volunteers. The MPM showed a decrease of the quantity of collagen and elastin as a function of depth of the dermis as well as age. The SAAID was lower for the older skin in the deeper dermis. Ultrasound imaging revealed a significant decrease of SENEB as a function of aging. The mechanical properties confirmed a loss of cutaneous elasticity and firmness. Although multiphoton microscopy is a powerful technique to study the characteristics of the dermis and its age-related damage, the location of the measurements (depth remains very important for the validation of these variations. These variations do not seem to be homogeneous according to the part of the dermis that is studied.

  7. Superficial nephrons in BALB/c and C57BL/6 mice facilitate in vivo multiphoton microscopy of the kidney.

    Directory of Open Access Journals (Sweden)

    Ina Maria Schießl

    Full Text Available Multiphoton microscopy (MPM offers a unique approach for addressing both the function and structure of an organ in near-real time in the live animal. The method however is limited by the tissue-specific penetration depth of the excitation laser. In the kidney, structures in the range of 100 µm from the surface are accessible for MPM. This limitation of MPM aggravates the investigation of the function of structures located deeper in the renal cortex, like the glomerulus and the juxtaglomerular apparatus. In view of the relevance of gene-targeted mice for investigating the function of these structures, we aimed to identify a mouse strain with a high percentage of superficially located glomeruli. The mean distance of the 30 most superficial glomeruli from the kidney surface was determined in 10 commonly used mouse strains. The mean depth of glomeruli was 118.4±3.4, 123.0±2.7, 133.7±3.0, 132.3±2.6, 141.0±4.0, 145.3±4.3, 148.9±4.2, 151.6±2.7, 167.7±3.9, and 207.8±3.2 µm in kidney sections from 4-week-old C3H/HeN, BALB/cAnN, SJL/J, C57BL/6N, DBA/2N, CD1 (CRI, 129S2/SvPas, CB6F1, FVB/N and NMRI (Han mice, respectively (n = 5 animals from each strain. The mean distance from the kidney surface of the most superficial glomeruli was significantly lower in the strains C3H/HeN Crl, BALB/cAnN, DBA/2NCrl, and C57BL/6N when compared to a peer group consisting of all the other strains (p<.0001. In 10-week-old mice, the most superficial glomeruli were located deeper in the cortex when compared to 4-week-old animals, with BALB/cAnN and C57BL/6N being the strains with the highest percentage of superficial glomeruli (25% percentile 116.7 and 121.9 µm, respectively. In summary, due to significantly more superficial glomeruli compared to other commonly used strains, BALB/cAnN and C57BL/6N mice appear to be particularly suitable for the investigation of glomerular function using MPM.

  8. Radioactive Substances Act 1960

    International Nuclear Information System (INIS)

    1960-01-01

    This Act regulates the keeping and use of radioactive material and makes provision for the disposal and storage of radioactive waste in the United Kingdom. It provides for a licensing system for such activities and for exemptions therefrom, in particular as concerns the United Kingdom Atomic Energy Authority. The Act repeals Section 4(5) of the Atomic Energy Authority Act, 1954 which made temporary provision for discharge of waste on or from premises occupied by the Authority. (NEA) [fr

  9. Learning dialog act processing

    OpenAIRE

    Wermter, Stefan; Löchel, Matthias

    1996-01-01

    In this paper we describe a new approach for learning dialog act processing. In this approach we integrate a symbolic semantic segmentation parser with a learning dialog act network. In order to support the unforeseeable errors and variations of spoken language we have concentrated on robust data-driven learning. This approach already compares favorably with the statistical average plausibility method, produces a segmentation and dialog act assignment for all utterances in a robust manner,...

  10. Multiphoton microscopy based cryo-imaging of inflated frozen human lung sections at -60°C in healthy and COPD lungs

    Science.gov (United States)

    Abraham, Thomas; Kayra, Damian; Zhang, Angela; Suzuki, Masaru; McDonough, John; Elliott, W. M.; Cooper, Joel D.; Hogg, James C.

    2013-02-01

    Lung is a complex gas exchanger with interfacial area (where the gas exchange takes place) is about the size of a tennis court. Respiratory function is linked to the biomechanical stability of the gas exchange or alveolar regions which directly depends on the spatial distributions of the extracellular matrix fibers such fibrillar collagens and elastin fibers. It is very important to visualize and quantify these fibers at their native and inflated conditions to have correct morphometric information on differences between control and diseased states. This can be only achieved in the ex vivo states by imaging directly frozen lung specimens inflated to total lung capacity. Multiphoton microscopy, which uses ultra-short infrared laser pulses as the excitation source, produces multiphoton excitation fluorescence (MPEF) signals from endogenously fluorescent proteins (e.g. elastin) and induces specific second harmonic generation (SHG) signals from non-centrosymmetric proteins such as fibrillar collagens in fresh human lung tissues [J. Struct. Biol. (2010)171,189-196]. Here we report for the first time 3D image data obtained directly from thick frozen inflated lung specimens (~0.7- 1.0 millimeter thick) visualized at -60°C without prior fixation or staining in healthy and diseased states. Lung specimens donated for transplantation and released for research when no appropriate recipient was identified served as controls, and diseased lung specimens donated for research by patients receiving lung transplantation for very severe COPD (n=4) were prepared as previously described [N. Engl. J. Med. (2011) 201, 1567]. Lung slices evenly spaced between apex and base were examined using multiphoton microscopy while maintained at -60°C using a temperature controlled cold stage with a temperature resolution of 0.1°C. Infrared femto-second laser pulses tuned to 880nm, dry microscopic objectives, and non-de-scanned detectors/spectrophotometer located in the reflection geometry were

  11. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... Reform Could Impact People With CF The Preserving Employee Wellness Programs Act Our Advocacy Work Advocacy Achievements Advocacy News Briefings, Testimonies, and Regulatory ...

  12. Radiological Protection Act 1970

    International Nuclear Information System (INIS)

    1970-01-01

    This Act provides for the establishment of a Radiological Protection Board to undertake research and advise on protection from radiation hazards. Its functions include provision of advice to Government departments with responsibilities in relation to protection of sectors of the community or the community as a whole against the hazards of ionizing radiation. The Act, which lays down that the Board shall replace certain departments concerned with radiation protection, repeals several Sections of the Radioactive Substances Act 1948 and the Science and Technology Act 1965. (NEA) [fr

  13. 25 CFR 700.33 - Act (The Act).

    Science.gov (United States)

    2010-04-01

    ... 25 Indians 2 2010-04-01 2010-04-01 false Act (The Act). 700.33 Section 700.33 Indians THE OFFICE OF NAVAJO AND HOPI INDIAN RELOCATION COMMISSION OPERATIONS AND RELOCATION PROCEDURES General Policies and Instructions Definitions § 700.33 Act (The Act). (a) The Act. The Act is Pub. L. 93-531, (88 Stat...

  14. Laser-Bioplasma Interaction: Excitation and Suppression of the Brain Waves by the Multi-photon Pulsed-operated Fiber Lasers in the Ultraviolet Range of Frequencies

    Science.gov (United States)

    Stefan, V. Alexander; IAPS-team Team

    2017-10-01

    The novel study of the laser excitation-suppression of the brain waves is proposed. It is based on the pulsed-operated multi-photon fiber-laser interaction with the brain parvalbumin (PV) neurons. The repetition frequency matches the low frequency brain waves (5-100 Hz); enabling the resonance-scanning of the wide range of the PV neurons (the generators of the brain wave activity). The tunable fiber laser frequencies are in the ultraviolet frequency range, thus enabling the monitoring of the PV neuron-DNA, within the 10s of milliseconds. In medicine, the method can be used as an ``instantaneous-on-off anesthetic.'' Supported by Nikola Tesla Labs, Stefan University.

  15. Physics of the Brain. Prevention of the Epileptic Seizures by the Multi-photon Pulsed-operated Fiber Lasers in the Ultraviolet Range of Frequencies.

    Science.gov (United States)

    Stefan, V. Alexander; IAPS Team

    The novel study of the epileptogenesis mechanisms is proposed. It is based on the pulsed-operated (amplitude modulation) multi-photon (frequency modulation) fiber-laser interaction with the brain epilepsy-topion (the epilepsy onset area), so as to prevent the excessive electrical discharge (epileptic seizure) in the brain. The repetition frequency, Ω, matches the low frequency (epileptic) phonon waves in the brain. The laser repetition frequency (5-100 pulses per second) enables the resonance-scanning of the wide range of the phonon (possible epileptic-to-be) activity in the brain. The tunable fiber laser frequencies, Δω (multi photon operation), are in the ultraviolet frequency range, thus enabling monitoring of the electrical charge imbalance (within the 10s of milliseconds), and the DNA-corruption in the epilepsy-topion, as the possible cause of the disease. Supported by Nikola Tesla Labs., Stefan University.

  16. Flexible digital signal processing architecture for narrowband and spread-spectrum lock-in detection in multiphoton microscopy and time-resolved spectroscopy.

    Science.gov (United States)

    Wilson, Jesse W; Park, Jong Kang; Warren, Warren S; Fischer, Martin C

    2015-03-01

    The lock-in amplifier is a critical component in many different types of experiments, because of its ability to reduce spurious or environmental noise components by restricting detection to a single frequency and phase. One example application is pump-probe microscopy, a multiphoton technique that leverages excited-state dynamics for imaging contrast. With this application in mind, we present here the design and implementation of a high-speed lock-in amplifier on the field-programmable gate array (FPGA) coprocessor of a data acquisition board. The most important advantage is the inherent ability to filter signals based on more complex modulation patterns. As an example, we use the flexibility of the FPGA approach to enable a novel pump-probe detection scheme based on spread-spectrum communications techniques.

  17. Silicon isotope separation utilizing infrared multiphoton dissociation of Si{sub 2}F{sub 6} irradiated with two-color CO{sub 2} laser light

    Energy Technology Data Exchange (ETDEWEB)

    Yokoyama, Atsushi; Ohba, Hironori; Hashimoto, Masashi [Japan Atomic Energy Research Inst., Tokai, Ibaraki (Japan). Tokai Research Establishment; Ishii, Takeshi; Ohya, Akio [Nuclear Development Corp., Tokai, Ibaraki (Japan); Arai, Shigeyoshi [Hill Research Co. Ltd., Tokyo (Japan)

    2002-08-01

    Silicon isotope separation has been done by utilizing the Infrared Multiphoton Dissociation (IRMPD) of Si{sub 2}F{sub 6} irradiated with two-color CO{sub 2} laser lights. The two-color excitation method improved the separation efficiency keeping the high enrichment factors. For example, 99.74% of {sup 28}Si was obtained at 49.63% dissociation of Si{sub 2}F{sub 6} after the simultaneous irradiation of 200 pulses with 966.23 cm{sup -1} photons (0.084 J/cm{sup 2}) and 954.55 cm{sup -1} photons (0.658 J/cm{sup 2}), while 2000 pulses were needed to obtain 99.35% of {sup 28}Si at 35.6% dissociation in the case of only one-color irradiation at 954.55 cm{sup -1} (0.97 J/cm{sup 2}). (author)

  18. Dynamics of NO2 dissociation. Study by resonance-enhanced multi-photon ionisation of energy distribution and of anisotropies in fragments

    International Nuclear Information System (INIS)

    Mons, Michel

    1988-01-01

    In this research thesis, the author reports the use of laser resonance-enhanced multi-photon ionization and of time-of-flight mass spectrometry for a detailed characterization of fragments produced by a photo-dissociation process. The author more particularly addressed the case of a NO 2 molecule excited at low energies above the dissociation threshold. In the first part, the author discusses issues and problems related to molecular photo-dissociation. In the second part, he presents the developed method and shows that the combined use of both techniques allows a precise characterisation of photo-fragments in terms of internal or translational energies as well as in terms of angle distributions. Finally, the author presents and discusses results obtained in the case of NO 2 [fr

  19. Detection of polychlorinated biphenyls in transformer oils in Vietnam by multiphoton ionization mass spectrometry using a far-ultraviolet femtosecond laser as an ionization source.

    Science.gov (United States)

    Duong, Vu Thi Thuy; Duong, Vu; Lien, Nghiem Thi Ha; Imasaka, Tomoko; Tang, Yuanyuan; Shibuta, Shinpei; Hamachi, Akifumi; Hoa, Do Quang; Imasaka, Totaro

    2016-03-01

    Polychlorinated biphenyls (PCBs) in transformer and food oils were measured using gas chromatography combined with multiphoton ionization mass spectroscopy. An ultrashort laser pulse emitting in the far-ultraviolet region was utilized for efficient ionization of the analytes. Numerous signal peaks were clearly observed for a standard sample mixture of PCBs when the third and fourth harmonic emissions (267 and 200nm) of a femtosecond Ti:sapphire laser (800nm) were employed. The signal intensities were found to be greater when measured at 200nm compared with those measured at 267nm, providing lower detection limits especially for highly chlorinated PCBs at shorter wavelengths. After simple pretreatment using disposable columns, PCB congeners were measured and found to be present in the transformer oils used in Vietnam. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. Silicon isotope separation utilizing infrared multiphoton dissociation of Si2F6 irradiated with two-color CO2 laser light

    International Nuclear Information System (INIS)

    Yokoyama, Atsushi; Ohba, Hironori; Hashimoto, Masashi; Arai, Shigeyoshi

    2002-01-01

    Silicon isotope separation has been done by utilizing the Infrared Multiphoton Dissociation (IRMPD) of Si 2 F 6 irradiated with two-color CO 2 laser lights. The two-color excitation method improved the separation efficiency keeping the high enrichment factors. For example, 99.74% of 28 Si was obtained at 49.63% dissociation of Si 2 F 6 after the simultaneous irradiation of 200 pulses with 966.23 cm -1 photons (0.084 J/cm 2 ) and 954.55 cm -1 photons (0.658 J/cm 2 ), while 2000 pulses were needed to obtain 99.35% of 28 Si at 35.6% dissociation in the case of only one-color irradiation at 954.55 cm -1 (0.97 J/cm 2 ). (author)

  1. Diamond-like-carbon nanoparticle production and agglomeration following UV multi-photon excitation of static naphthalene/helium gas mixtures

    Energy Technology Data Exchange (ETDEWEB)

    Walsh, A. J.; Ruth, A. A., E-mail: a.ruth@ucc.ie [Physics Department and Environmental Research Institute, University College Cork, Cork (Ireland); Tielens, A. G. G. M. [Leiden Observatory, Leiden University, Niels Bohrweg 2, 2333-CA Leiden (Netherlands)

    2016-07-14

    We report the formation of nanoparticles with significant diamond character after UV multi-photon laser excitation of gaseous naphthalene, buffered in static helium gas, at room temperature. The nanoparticles are identified in situ by their absorption and scattering spectra between 400 and 850 nm, which are modeled using Mie theory. Comparisons of the particles’ spectroscopic and optical properties with those of carbonaceous materials indicate a sp{sup 3}/sp{sup 2} hybridization ratio of 8:1 of the particles formed. The particle extinction in the closed static (unstirred) gas-phase system exhibits a complex and quasi-oscillatory time dependence for the duration of up to several hours with periods ranging from seconds to many minutes. The extinction dynamics of the system is based on a combination of transport features and particle interaction, predominantly agglomeration. The relatively long period of agglomeration allows for a unique analysis of the agglomeration process of diamond-like carbon nanoparticles in situ.

  2. Marine Mammal Protection Act

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The Marine Mammal Protection Act (MMPA or Act) prohibits, with certain exceptions, the "take" of marine mammals in U.S. waters and by U.S. citizens on the high seas,...

  3. Collective speech acts

    NARCIS (Netherlands)

    Meijers, A.W.M.; Tsohatzidis, S.L.

    2007-01-01

    From its early development in the 1960s, speech act theory always had an individualistic orientation. It focused exclusively on speech acts performed by individual agents. Paradigmatic examples are ‘I promise that p’, ‘I order that p’, and ‘I declare that p’. There is a single speaker and a single

  4. Americans with Disabilities Act.

    Science.gov (United States)

    Kaesberg, Mary Ann; Murray, Kenneth T.

    1994-01-01

    Presents a 35-item checklist of practical activities for school district compliance with the Americans with Disabilities Act (ADA). The checklist is based on ADA statutes, other civil rights legislation and litigation, as well as pertinent regulations and the legislative history of the act contained in the Congressional Record. (MLF)

  5. Radiation emitting devices act

    International Nuclear Information System (INIS)

    1970-01-01

    This Act, entitled the Radiation Emitting Devices Act, is concerned with the sale and importation of radiation emitting devices. Laws relating to the sale, lease or import, labelling, advertising, packaging, safety standards and inspection of these devices are listed as well as penalties for any person who is convicted of breaking these laws

  6. Nuclear Installations Act 1969

    International Nuclear Information System (INIS)

    1969-01-01

    The purpose of this Act is to amend the Nuclear Installations Act 1965 to bring it into full compliance with the international conventions on nuclear third party liability to which the United Kingdom is a Signatory, namely, the Paris Convention, the Brussels Supplementary Convention and the Vienna Convention. (NEA) [fr

  7. Cellular organization and spectral diversity of GFP-like proteins in live coral cells studied by single and multiphoton imaging and microspectroscopy

    Science.gov (United States)

    Salih, Anya; Cox, Guy C.; Larkum, Anthony W.

    2003-07-01

    Tissues of many marine invertebrates of class Anthozoa contain intensely fluorescent or brightly coloured pigments. These pigments belong to a family of photoactive proteins closely related to Green Fluorescent Protein (GFP), and their emissions range from blue to red wavelengths. The great diversity of these pigments has only recently been realised. To investigate the role of these proteins in corals, we have performed an in vivo fluorescent pigment (FP) spectral and cellular distribution analyses in live coral cells using single and multi-photon laser scanning imaging and microspectroscopy. These analyses revealed that even single colour corals contain spectroscopically heterogeneous pigment mixtures, with 2-5 major colour types in the same area of tissue. They were typically arranged in step-wise light emission energy gradients (e.g. blue, green, yellow, red). The successive overlapping emission-excitation spectral profiles of differently coloured FPs suggested that they were suited for sequential energy coupling. Traces of red FPs (emission = 570-660 nm) were present, even in non-red corals. We confirmed that radiative energy transfer could occur between separate granules of blue and green FPs and that energy transfer was inversely proportional to the square of the distance between them. Multi-photon micro-spectrofluorometric analysis gave significantly improved spectral resolution by restricting FP excitation to a single point in the focal plane of the sample. Pigment heterogeneity at small scales within granules suggested that fluorescence resonance energy transfer (FRET) might be occurring, and we confirmed that this was the case. Thus, energy transfer can take place both radiatively and by FRET, probably functioning in photoprotection by dissipation of excessive solar radiation.

  8. Multi-photon entanglements

    International Nuclear Information System (INIS)

    Daniell, M.L.

    2000-09-01

    The motivation of this thesis was to create higher-order entanglements. The first experimental observation of a four-photon entanglement was presented in the experiment of this thesis. And the visibility of this entanglement was 0.79+-0.06, which is sufficient to make claims of the nonlocality of quantum mechanics. This therefore lays a foundation for experiments showing the nonlocality of teleportation, and the purification of entanglement. The work of this thesis brings together a lot of earlier work done by the Zeilinger Group, and lays a foundation for future experiments. Earlier experiments such as teleportation together with entanglement swapping, which are 'complete teleportation' in as much as the state teleported is entirely undefined, can be combined and re-done with this four-photon entanglement. This result would be the first demonstration of complete, nonlocal teleportation. Also this experiment can be slightly modified and used to perform the first experimental quantum purification of entanglement, which is of vital importance to the fields of quantum information, and also is interesting for fundamental experiments on entanglement. Another direct application of this experiment is to perform the first 'event-ready' testing of Bell's Inequality. Here the four-photon entanglement can be used as a source of entangled photons, whereby the photons have no common source. This would enable an even more stringent testing of Bells theorem. Finally this experiment can be used for the demonstration and investigation of many practical, directly applicable quantum information schemes. For instance quantum cryptography, error correction, and computing. (author)

  9. Multiphoton polarization Bremsstrahlung effect

    International Nuclear Information System (INIS)

    Golovinskij, P.A.

    2001-01-01

    A general approach to induced polarization effects was formulated on the basis of theory of many particles in a strong periodic field. Correlation with the perturbation theory is shown and the types of effective polarization potentials both for isolated atoms and ions, and for ions in plasma, are provided. State of art in the theory of forced polarization Bremsstrahlung effect is analyzed and some outlooks for further experimental and theoretical studies are outlined [ru

  10. The Radiation Protection Act

    International Nuclear Information System (INIS)

    Persson, L.

    1989-01-01

    The new Radiation Protection Act (1988:220) entered into force in Sweden on July 1st, 1988. This book presents the Act as well as certain regulations connected to it. As previously, the main responsibility for public radiation protection will rest with one central radiation protection authority. According to the 1988 Act, the general obligations with regard to radiation protection will place a greater responsibility than in the past on persons carrying out activities involving radiation. Under the act, it is possible to adjust the licensing and supervisory procedures to the level of danger of the radiation source and the need for adequate competence, etc. The Act recognises standardised approval procedures combined with technical regulations for areas where the risks are well known. The Act contains several rules providing for more effective supervision. The supervising authority may in particular decide on the necessary regulations and prohibitions for each individual case. The possibilities of using penal provisions have been extended and a rule on the mandatory execution of orders has been introduced. The Ordinance on Radiation Protection (1988:293) designates the National Institute of Radiation Protection (SSI) as the central authority referred to in the Radiation Protection Act. The book also gives a historic review of radiation protection laws in Sweden, lists regulations issued by SSI and presents explanations of radiation effects and international norms in the area. (author)

  11. Canada's Clean Air Act

    International Nuclear Information System (INIS)

    2006-01-01

    This paper provided an outline of Canada's Clean Air Act and examined some of the regulatory changes that will occur as a result of its implementation. The Act is being introduced to strengthen the legislative basis for taking action on reducing air pollution and GHGs, and will allow the government to regulate both indoor and outdoor air pollutants and GHGs. The Act will require the Ministers of the Environment and Health to establish national air quality objectives, as well as to monitor and report on their attainment. The Canadian Environmental Protection Act will be amended to enable the government to regulate the blending of fuels and their components. The Motor Vehicle Fuel Consumption Standards Act will also be amended to enhance the government's authority to regulate vehicle fuel efficiency. The Energy Efficiency Act will also be expanded to allow the government to set energy efficiency standards and labelling requirements for a wider range of consumer and commercial products. The Act will commit to short, medium and long-term industrial air pollution targets. Regulations will be proposed for emissions from industry; on-road and off-road vehicles and engines; and consumer and commercial products. It was concluded that the Government of Canada will continue to consult with provinces, territories, industries and Canadians to set and reach targets for the reduction of both indoor and outdoor air pollutants and GHG emissions. 6 figs

  12. The Energy Act 1983

    International Nuclear Information System (INIS)

    1983-01-01

    Part II of This Act came into force on 1 September 1983 and is concerned with nuclear installations. Its main purpose is to amend the Nuclear Installations Act 1965 to give effect to the provisions of two Protocols amending the Paris Convention on nuclear third party liability and the Brussels Convention Supplementary to the Paris Convention respectively. The principal effect of these modifications is to increase the sums available to meet claims for nuclear damage. The United Kingdom is a Party to both Conventions and the provisions of the 1983 Act will enable it to ratify the Protocols. (NEA) [fr

  13. 75 FR 63703 - Privacy Act of 1974; Privacy Act Regulation

    Science.gov (United States)

    2010-10-18

    ... FEDERAL RESERVE SYSTEM 12 CFR Part 261a [Docket No. R-1313] Privacy Act of 1974; Privacy Act... implementing the Privacy Act of 1974 (Privacy Act). The primary changes concern the waiver of copying fees... records under the Privacy Act; the amendment of special procedures for the release of medical records to...

  14. Abandoned Shipwreck Act

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This data represents the extent of the Abandoned Shipwreck Act (ASA). The ASA allows states to manage a broad range of resources within submerged lands, including...

  15. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... The Preserving Employee Wellness Programs Act Our Advocacy Work Advocacy ... the value of tapping into the expertise that only people with CF and their families have. We invite you to share insights to ...

  16. Energy Policy Act

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The Energy Policy Act (EPA) addresses energy production in the United States, including: (1) energy efficiency; (2) renewable energy; (3) oil and gas; (4) coal; (5)...

  17. ACTS – SUCCESS STORY

    Indian Academy of Sciences (India)

    First page Back Continue Last page Overview Graphics. ACTS – SUCCESS STORY. Totally 103 experiments were conducted and the programme succeeded in the areas. Medicine; Education; Defence; Emergency Response; Maritime and Aeronautical Mobile Communications; Science and Astronomy.

  18. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... There are different ways to clear your airways. Most are easy to do. Infants and toddlers will ... best ACT is the one that you are most likely to perform as part of your daily ...

  19. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... to loosen mucus from airway walls. See how different airway clearance techniques work to help you clear the thick, sticky mucus ... Offer their tips for fitting ACTs into daily life Airway Clearance Techniques | Webcast ... Facebook Twitter ...

  20. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... CF Community in Health Care Reform Milestones in Health Care Reform How Tax Reform Could Impact People With CF The Preserving Employee Wellness Programs Act Our Advocacy Work Advocacy Achievements ...

  1. Clean Water Act

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — These data represent geographic terms used within the Clean Water Act (CWA). The CWA establishes the basic structure for regulating the addition of pollutants...

  2. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... toddlers will need help from a parent or caregiver. Older kids and adults can choose ACTs that ... into the smaller airways to attack bacteria. Choose What's Best for You Your respiratory therapist or another ...

  3. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... that help thin and move the mucus, and antibiotics. Bronchodilators should be inhaled before you start ACTs. This medication helps to widen your airways (bronchi) by relaxing the ...

  4. Endangered Species Act

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The purpose of the Endangered Species Act (ESA) is to protect and recover imperiled species and the ecosystems upon which they depend. The U.S. Fish and Wildlife...

  5. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... Reform Could Impact People With CF The Preserving Employee Wellness Programs Act Our Advocacy Work Advocacy Achievements ... on their own. Share Facebook Twitter Email More options Print Share Facebook Twitter Email Print Permalink All ...

  6. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... 3-D Structure Consortium CFTR Folding Consortium Epithelial Stem Cell Consortium Mucociliary Clearance Consortium SUCCESS WITH THERAPIES RESEARCH ... ACTs involve coughing or huffing . Many of them use percussion (clapping) or vibration to loosen mucus from ...

  7. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... today. ANNUAL FUND Become a Corporate Supporter Cause Marketing Make a Charitable Gift Our Corporate Supporters Workplace ... Clearance Airway Clearance Techniques (ACTs) There are different ways to clear your airways. Most are easy to ...

  8. Marine Pollution Prevention Act

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The Marine Pollution Prevention Act of 2008 implements the International Convention for the Prevention of Pollution from Ships, including related Protocols (MARPOL)...

  9. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... CF Community in Health Care Reform Milestones in Health Care Reform How Tax Reform Could Impact People With CF The Preserving Employee Wellness Programs Act Our Advocacy Work Advocacy Achievements Advocacy News Briefings, Testimonies, and Regulatory ...

  10. Preventive Radiation Protection Act

    International Nuclear Information System (INIS)

    Roewer, H.

    1988-01-01

    The commentary is intended to contribute to protection of the population by a practice-oriented discussion and explanation of questions arising in connection with the Preventive Radiation Protection Act. Leaving aside discussions about abandonment of nuclear power, or criticism from any legal point of view, the commentary adopts the practical approach that accepts, and tries to help implementing, the act as it is. It is a guide for readers who are not experts in the law and gives a line of orientation by means of explanations and sometimes by citations from other acts (in footnotes). The commentary also presents the EURATOM Directive No. 3954/87 dated 22 December 1987, the EC Directive No. 3955/87 dated 22 December 1987, and the EC Directive No. 1983/88 dated 5 July 1988. A tabular survey shows the system of duties and competences defined by the Preventive Radiation Protection Act. (RST) [de

  11. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... Contraception and Protection How Does CF Affect the Female Reproductive System? How Does CF Affect the Male ... Reform Could Impact People With CF The Preserving Employee Wellness Programs Act Our Advocacy Work Advocacy Achievements ...

  12. ACT250 Districts

    Data.gov (United States)

    Vermont Center for Geographic Information — The ACT 250 Districts layer is part of a larger dataset that contains administrative boundaries for Vermont's Agency of Natural Resources. The dataset includes...

  13. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... ACTs involve coughing or huffing . Many of them use percussion (clapping) or vibration to loosen mucus from airway walls. See how different airway clearance techniques work to help you clear the thick, sticky mucus ...

  14. Acts of Research

    DEFF Research Database (Denmark)

    Nelund, Sidsel

    place mostly in seminars and articles, in which knowledge is often discussed as an intrinsic quality of the artwork. Acts of Research, however, is devoted to studying the rise of knowledge production in contemporary art from the perspective of artistic, curatorial and educational research...... described as knowledge producers and exhibitions and art works as instances of knowledge production. Acts of Research: Knowledge Production in Contemporary Arts between Knowledge Economy and Critical Practices analyses this development. The academic discussion of knowledge production in the arts has taken...... with an awareness of larger political, economic, geographical and art-related aspects. The concept of ‘acts of research’ is suggested as a way to understand knowledge production as a creative act in which research carried out in relation to a specific material challenges and resists the protocols of conventional...

  15. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... Congressional Cystic Fibrosis Caucus Our Policy Agenda Policy Principles SIGN UP FOR ADVOCACY ACTION ALERTS Community We ... options Print Share Facebook Twitter Email Print Permalink All ACTs involve coughing or huffing . Many of them ...

  16. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... How Does CF Affect the Male Reproductive System? Sex and CF: Some Practical Advice Family Planning and ... need help from a parent or caregiver. Older kids and adults can choose ACTs that they can ...

  17. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... FOR ADVOCACY ACTION ALERTS Community We recognize the value of tapping into the expertise that only people ... ACTs involve coughing or huffing . Many of them use percussion (clapping) or vibration to loosen mucus from ...

  18. National Environmental Policy Act

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The National Environmental Policy Act (NEPA) was the first major environmental law in the United States and established national environmental policies for the...

  19. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... Reform Could Impact People With CF The Preserving Employee Wellness Programs Act Our Advocacy Work Advocacy Achievements ... Board of Trustees Our Leadership Careers Reports and Financials Contact Us Governance and Policies What is CF? ...

  20. Airway Clearance Techniques (ACTs)

    Medline Plus

    Full Text Available ... own. Share Facebook Twitter Email More options Print Share Facebook Twitter Email Print Permalink All ACTs involve ... CF Care Team Research About Our Research Developing New Treatments Researcher Resources Assistance Services Find Resources: CF ...