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Sample records for access multiphoton microscopy

  1. Multiphoton microscopy in neuroscience

    Denk, Winfried

    2002-06-01

    The study of the nervous system requires to an exceptional extent observation of and experimentation on intact tissue. There, in particular, high-resolution optical microscopy benefits from the inherent advantages of multi-photon fluorescence excitation. Several cases will be presented from a number of different tissues and organisms, where multi-photon excited laser scanning fluorescence microscopy has been an essential experimental tool. Those examples include the discovery of biochemical coincidence detection in synaptic spines and the clarification of the underlying mechanism; the observation of sensory evoked dendritic signaling in intact animals and the observation of light induced calcium signals in the intact retina. Recently a fiber coupled two-photon microscopy has been developed that allows the imaging in moving animal.

  2. Phase modulated multiphoton microscopy

    Karki, Khadga Jung; Pullerits, Tonu

    2015-01-01

    We show that the modulation of the phases of the laser beams of ultra-short pulses leads to modulation of the two photon fluorescence intensity. The phase modulation technique when used in multi-photon microscopy can improve the signal to noise ratio. The technique can also be used in multiplexing the signals in the frequency domain in multi-focal raster scanning microscopy. As the technique avoids the use of array detectors as well as elaborate spatiotemporal multiplexing schemes it provides a convenient means to multi-focal scanning in axial direction. We show examples of such uses. Similar methodology can be used in other non-linear scanning microscopies, such as second or third harmonic generation microscopy.

  3. Multi-photon excitation microscopy

    Faretta Mario; Vicidomini Giuseppe; Bianchini Paolo; Diaspro Alberto; Ramoino Paola; Usai Cesare

    2006-01-01

    Abstract Multi-photon excitation (MPE) microscopy plays a growing role among microscopical techniques utilized for studying biological matter. In conjunction with confocal microscopy it can be considered the imaging workhorse of life science laboratories. Its roots can be found in a fundamental work written by Maria Goeppert Mayer more than 70 years ago. Nowadays, 2PE and MPE microscopes are expected to increase their impact in areas such biotechnology, neurobiology, embryology, tissue engine...

  4. Differential Multiphoton Laser Scanning Microscopy

    Field, Jeffrey J.; Sheetz, Kraig E.; Chandler, Eric V.; Hoover, Erich E.; Young, Michael D.; Ding, Shi-you; Sylvester, Anne W.; Kleinfeld, David; Squier, Jeff A.

    2016-01-01

    Multifocal multiphoton microscopy (MMM) in the biological and medical sciences has become an important tool for obtaining high resolution images at video rates. While current implementations of MMM achieve very high frame rates, they are limited in their applicability to essentially those biological samples that exhibit little or no scattering. In this paper, we report on a method for MMM in which imaging detection is not necessary (single element point detection is implemented), and is therefore fully compatible for use in imaging through scattering media. Further, we demonstrate that this method leads to a new type of MMM wherein it is possible to simultaneously obtain multiple images and view differences in excitation parameters in a single shot.

  5. Multiphoton microscopy: An introduction to gastroenterologists

    Hye Jin Cho; Hoon Jai Chun; Eun Sun Kim; Bong Rae Cho

    2011-01-01

    Multiphoton microscopy, relying on the simultaneous absorption of two or more photons by a fluorophore, has come to occupy a prominent place in modern biomedical research with its ability to allow real-time observation of a single cell and molecules in intact tissues. Multiphoton microscopy exhibits nonlinear optical contrast properties, which can make it possible to provide an exceptionally large depth penetration with less phototoxicity. This system becomes more and more an inspiring tool for a non-invasive imaging system to realize "optical biopsy" and to examine the functions of living cells. In this review, we briefly present the physical principles and properties of multiphoton microscopy as well as the current applications in biological fields. In addition, we address what we see as the future potential of multiphoton microscopy for gastroenterologic research.

  6. Multiphoton microscopy in defining liver function

    Thorling, Camilla A.; Crawford, Darrell; Burczynski, Frank J.; Liu, Xin; Liau, Ian; Roberts, Michael S.

    2014-09-01

    Multiphoton microscopy is the preferred method when in vivo deep-tissue imaging is required. This review presents the application of multiphoton microscopy in defining liver function. In particular, multiphoton microscopy is useful in imaging intracellular events, such as mitochondrial depolarization and cellular metabolism in terms of NAD(P)H changes with fluorescence lifetime imaging microscopy. The morphology of hepatocytes can be visualized without exogenously administered fluorescent dyes by utilizing their autofluorescence and second harmonic generation signal of collagen, which is useful in diagnosing liver disease. More specific imaging, such as studying drug transport in normal and diseased livers are achievable, but require exogenously administered fluorescent dyes. If these techniques can be translated into clinical use to assess liver function, it would greatly improve early diagnosis of organ viability, fibrosis, and cancer.

  7. Clinical multiphoton and CARS microscopy

    Breunig, H. G.; Weinigel, M.; Darvin, M. E.; Lademann, J.; König, K.

    2012-03-01

    We report on clinical CARS imaging of human skin in vivo with the certified hybrid multiphoton tomograph CARSDermaInspect. The CARS-DermaInspect provides simultaneous imaging of non-fluorescent intradermal lipid and water as well as imaging of two-photon excited fluorescence from intrinsic molecules. Two different excitation schemes for CARS imaging have been realized: In the first setup, a combination of fs oscillator and optical parametric oscillator provided fs-CARS pump and Stokes pulses, respectively. In the second setup a fs oscillator was combined with a photonic crystal fiber which provided a broadband spectrum. A spectral range out of the broadband-spectrum was selected and used for CARS excitation in combination with the residual fs-oscillator output. In both setups, in addition to CARS, single-beam excitation was used for imaging of two-photon excited fluorescence and second harmonic generation signals. Both CARS-excitation systems were successfully used for imaging of lipids inside the skin in vivo.

  8. Multiphoton microscopy with near infrared contrast agents

    Yazdanfar, Siavash; Joo, Chulmin; Zhan, Chun; Berezin, Mikhail Y.; Akers, Walter J.; Achilefu, Samuel

    2010-05-01

    While multiphoton microscopy (MPM) has been performed with a wide range of excitation wavelengths, fluorescence emission has been limited to the visible spectrum. We introduce a paradigm for MPM of near-infrared (NIR) fluorescent molecular probes via nonlinear excitation at 1550 nm. This all-NIR system expands the range of available MPM fluorophores, virtually eliminates background autofluorescence, and allows for use of fiber-based, turnkey ultrafast lasers developed for telecommunications.

  9. Pulse front adaptive optics in multiphoton microscopy

    Sun, B.; Salter, P. S.; Booth, M. J.

    2016-03-01

    The accurate focusing of ultrashort laser pulses is extremely important in multiphoton microscopy. Using adaptive optics to manipulate the incident ultrafast beam in either the spectral or spatial domain can introduce significant benefits when imaging. Here we introduce pulse front adaptive optics: manipulating an ultrashort pulse in both the spatial and temporal domains. A deformable mirror and a spatial light modulator are operated in concert to modify contours of constant intensity in space and time within an ultrashort pulse. Through adaptive control of the pulse front, we demonstrate an enhancement in the measured fluorescence from a two photon microscope.

  10. Optimizing Fluorescence Collection Efficiency in Multiphoton Microscopy

    Zinter, Joseph P.

    Over the past 20 years multiphoton microscopy has established itself as the premier modality for high resolution (techniques result from the use of longer wavelength excitation light (˜700--1000 nm), enabling deeper tissue penetration, and the spatially and temporally localized generation of fluorescence inherent in two-photon excitation, which virtually eliminates out-of-focus fluorescence. Imaging depths of ˜500 mum are now considered common practice, however, as the technique continues to increase in popularity and utility significant efforts are being made to maximize imaging depth. These efforts can be compartmentalized into those aimed at increasing fluorescence excitation and those attempting to maximize fluorescence collection. Progress has been made in increasing fluorescence excitation efficiency, however these approaches require costly and complicated optical instrumentation, and are often deemed impractical. Efforts targeted at increasing fluorescence collection efficiency have been limited due to the complexity of describing the ensemble of scattered fluorescent photons emerging from a sample and propagating through a microscope objective and the subsequent fluorescence collection pathway of a multiphoton microscope. Since there is no analytic solution for these fluorescent photon distributions as a function of imaging depth in the relevant, non-diffusive multiphoton imaging range, numerical techniques are required. Here is presented the first computational model of fluorescence propagation through the complete fluorescence collection pathway of a multiphoton microscope. Monte Carlo simulations were used to model the propagation of fluorescence as a function of imaging depth in a scattering sample with physiologically accurate optical properties, and to determine the collected and transmitted fluorescent fraction through the Olympus 20X 0.95NA microscope objective, one of the highest performance and most commonly used objectives for multiphoton

  11. Multimodal optoacoustic and multiphoton fluorescence microscopy

    Sela, Gali; Razansky, Daniel; Shoham, Shy

    2013-03-01

    Multiphoton microscopy is a powerful imaging modality that enables structural and functional imaging with cellular and sub-cellular resolution, deep within biological tissues. Yet, its main contrast mechanism relies on extrinsically administered fluorescent indicators. Here we developed a system for simultaneous multimodal optoacoustic and multiphoton fluorescence 3D imaging, which attains both absorption and fluorescence-based contrast by integrating an ultrasonic transducer into a two-photon laser scanning microscope. The system is readily shown to enable acquisition of multimodal microscopic images of fluorescently labeled targets and cell cultures as well as intrinsic absorption-based images of pigmented biological tissue. During initial experiments, it was further observed that that detected optoacoustically-induced response contains low frequency signal variations, presumably due to cavitation-mediated signal generation by the high repetition rate (80MHz) near IR femtosecond laser. The multimodal system may provide complementary structural and functional information to the fluorescently labeled tissue, by superimposing optoacoustic images of intrinsic tissue chromophores, such as melanin deposits, pigmentation, and hemoglobin or other extrinsic particle or dye-based markers highly absorptive in the NIR spectrum.

  12. Multifocal multiphoton microscopy based on multianode photomultiplier tubes

    Kim, Ki Hean; Buehler, Christof; Bahlmann, Karsten; Ragan, Timothy; Lee, Wei-Chung A.; Nedivi, Elly; Heffer, Erica L.; Fantini, Sergio; So, Peter T. C.

    2007-01-01

    Multifocal multiphoton microscopy (MMM) enhances imaging speed by parallelization. It is not well understood why the imaging depth of MMM is significantly shorter than conventional single-focus multiphoton microscopy (SMM). In this report, we show that the need for spatially resolved detectors in MMM results in a system that is more sensitive to the scattering of emission photons with reduced imaging depth. For imaging depths down to twice the scattering mean free path length of emission phot...

  13. Multiphoton spectral microscopy for imaging and quantification of tissue glycation

    Tseng, Jo-Ya; Ghazaryan, Ara A.; Lo, Wen; Chen, Yang-Fang; Hovhannisyan, Vladimir; Chen, Shean-Jen; Tan, Hsin-Yuan; Dong, Chen-Yuan

    2010-01-01

    Tissue glycation from diabetes and aging can result in complications such as renal failure, blindness, nerve damage and vascular diseases. In this work, we applied multiphoton microscopy for imaging and characterizing the extent of tissue glycation. The characteristic features of multiphoton autofluorescence (MPAF) and second harmonic generation (SHG) images as well as MPAF spectra of glycated bovine skin, cornea and aorta were acquired. The analysis of MPAF intensity change accompanying the ...

  14. Multicolor multiphoton microscopy based on a nanosecond supercontinuum laser source.

    Lefort, Claire; O'Connor, Rodney P; Blanquet, Véronique; Magnol, Laetitia; Kano, Hideaki; Tombelaine, Vincent; Lévêque, Philippe; Couderc, Vincent; Leproux, Philippe

    2016-07-01

    Multicolor multiphoton microscopy is experimentally demonstrated for the first time on a spectral bandwidth of excitation of 300 nm (full width half maximum) thanks to the implementation a nanosecond supercontinuum (SC) source compact and simple with a low repetition rate. The interest of such a wide spectral bandwidth, never demonstrated until now, is highlighted in vivo: images of glioma tumor cells stably expressing eGFP grafted on the brain of a mouse and its blood vessels network labelled with Texas Red(®) are obtained. These two fluorophores have a spectral bandwidth covering the whole 300 nm available. In parallel, a similar image quality is obtained on a sample of mouse muscle in vitro when excited with this nanosecond SC source or with a classical high rate, femtosecond and quasi monochromatic laser. This opens the way for (i) a simple and very complete biological characterization never performed to date with multiphoton processes, (ii) multiple means of contrast in nonlinear imaging allowed by the use of numerous fluorophores and (iii) other multiphoton processes like three-photon ones. PMID:26872004

  15. Microstructure imaging of human rectal mucosa using multiphoton microscopy

    Liu, N R; Chen, J X; Zhuo, S M; Zheng, L Q; Jiang, X S [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 (China); Chen, G [Department of Pathology, Fujian Provincial Tumor Hospital, Fuzhou (China); Yan, J, E-mail: chenjianxin@fjnu.edu.cn, E-mail: ynjun@yahoo.com [Department of Surgery, Fujian Provincial Tumor Hospital, Fuzhou (China)

    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.

  16. Reassignment of scattered emission photons in multifocal multiphoton microscopy.

    Cha, Jae Won; Singh, Vijay Raj; Kim, Ki Hean; Subramanian, Jaichandar; Peng, Qiwen; Yu, Hanry; Nedivi, Elly; So, Peter T C

    2014-01-01

    Multifocal multiphoton microscopy (MMM) achieves fast imaging by simultaneously scanning multiple foci across different regions of specimen. The use of imaging detectors in MMM, such as CCD or CMOS, results in degradation of image signal-to-noise-ratio (SNR) due to the scattering of emitted photons. SNR can be partly recovered using multianode photomultiplier tubes (MAPMT). In this design, however, emission photons scattered to neighbor anodes are encoded by the foci scan location resulting in ghost images. The crosstalk between different anodes is currently measured a priori, which is cumbersome as it depends specimen properties. Here, we present the photon reassignment method for MMM, established based on the maximum likelihood (ML) estimation, for quantification of crosstalk between the anodes of MAPMT without a priori measurement. The method provides the reassignment of the photons generated by the ghost images to the original spatial location thus increases the SNR of the final reconstructed image. PMID:24898470

  17. Reassignment of Scattered Emission Photons in Multifocal Multiphoton Microscopy

    Cha, Jae Won; Singh, Vijay Raj; Kim, Ki Hean; Subramanian, Jaichandar; Peng, Qiwen; Yu, Hanry; Nedivi, Elly; So, Peter T. C.

    2014-06-01

    Multifocal multiphoton microscopy (MMM) achieves fast imaging by simultaneously scanning multiple foci across different regions of specimen. The use of imaging detectors in MMM, such as CCD or CMOS, results in degradation of image signal-to-noise-ratio (SNR) due to the scattering of emitted photons. SNR can be partly recovered using multianode photomultiplier tubes (MAPMT). In this design, however, emission photons scattered to neighbor anodes are encoded by the foci scan location resulting in ghost images. The crosstalk between different anodes is currently measured a priori, which is cumbersome as it depends specimen properties. Here, we present the photon reassignment method for MMM, established based on the maximum likelihood (ML) estimation, for quantification of crosstalk between the anodes of MAPMT without a priori measurement. The method provides the reassignment of the photons generated by the ghost images to the original spatial location thus increases the SNR of the final reconstructed image.

  18. Superresolved multiphoton microscopy with spatial frequency-modulated imaging.

    Field, Jeffrey J; Wernsing, Keith A; Domingue, Scott R; Allende Motz, Alyssa M; DeLuca, Keith F; Levi, Dean H; DeLuca, Jennifer G; Young, Michael D; Squier, Jeff A; Bartels, Randy A

    2016-06-14

    Superresolved far-field microscopy has emerged as a powerful tool for investigating the structure of objects with resolution well below the diffraction limit of light. Nearly all superresolution imaging techniques reported to date rely on real energy states of fluorescent molecules to circumvent the diffraction limit, preventing superresolved imaging with contrast mechanisms that occur via virtual energy states, including harmonic generation (HG). We report a superresolution technique based on spatial frequency-modulated imaging (SPIFI) that permits superresolved nonlinear microscopy with any contrast mechanism and with single-pixel detection. We show multimodal superresolved images with two-photon excited fluorescence (TPEF) and second-harmonic generation (SHG) from biological and inorganic media. Multiphoton SPIFI (MP-SPIFI) provides spatial resolution up to 2η below the diffraction limit, where η is the highest power of the nonlinear intensity response. MP-SPIFI can be used to provide enhanced resolution in optically thin media and may provide a solution for superresolved imaging deep in scattering media. PMID:27231219

  19. Superresolved multiphoton microscopy with spatial frequency-modulated imaging

    Field, Jeffrey J.; Wernsing, Keith A.; Domingue, Scott R.; Allende Motz, Alyssa M.; DeLuca, Keith F.; Levi, Dean H.; DeLuca, Jennifer G.; Young, Michael D.; Squier, Jeff A.; Bartels, Randy A.

    2016-05-26

    Superresolved far-field microscopy has emerged as a powerful tool for investigating the structure of objects with resolution well below the diffraction limit of light. Nearly all superresolution imaging techniques reported to date rely on real energy states of fluorescent molecules to circumvent the diffraction limit, preventing superresolved imaging with contrast mechanisms that occur via virtual energy states, including harmonic generation (HG). We report a superresolution technique based on spatial frequency-modulated imaging (SPIFI) that permits superresolved nonlinear microscopy with any contrast mechanism and with single-pixel detection. We show multimodal superresolved images with two-photon excited fluorescence (TPEF) and second-harmonic generation (SHG) from biological and inorganic media. Multiphoton SPIFI (MP-SPIFI) provides spatial resolution up to 2..eta.. below the diffraction limit, where ..eta.. is the highest power of the nonlinear intensity response. MP-SPIFI can be used to provide enhanced resolution in optically thin media and may provide a solution for superresolved imaging deep in scattering media.

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

    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.

  1. Characterization of powdered epidermal vaccine delivery with multiphoton microscopy

    Multiphoton laser scanning microscopy (MPLSM) has been adapted to non-invasively characterize hand-held powdered epidermal vaccine delivery technology. A near infrared femtosecond pulsed laser, wavelength at approximately 920 nm, was used to evoke autofluorescence of endogenous fluorophores within ex vivo porcine and human skin. Consequently, sub cellular resolution three-dimensional images of stratum corneum and viable epidermal cells were acquired and utilized to observe the morphological deformation of these cells as a result of micro-particle penetration. Furthermore, the distributional pattern of micro-particles within the specific skin target volume was quantified by measuring the penetration depth as revealed by serial optical sections in the axial plane obtained with MPLSM. Additionally, endogenous fluorescence contrast images acquired at the supra-basal layer reveal cellular structures that may pertain to dendritic Langerhans cells of the epidermis. These results show that MPLSM has advantages over conventional histological approaches, since three-dimensional functional images with sub-cellular spatial resolution to depths beyond the epidermis can be acquired non-invasively. Accordingly, we propose that MPLSM is ideal for investigations of powdered epidermal vaccine delivery

  2. Simultaneous imaging of GFP, CFP and collagen in tumors in vivo using multiphoton microscopy

    Segall Jeffrey E

    2005-05-01

    Full Text Available Abstract Background The development of multiphoton laser scanning microscopy has greatly facilitated the imaging of living tissues. However, the use of genetically encoded fluorescent proteins to distinguish different cell types in living animals has not been described at single cell resolution using multiphoton microscopy. Results Here we describe a method for the simultaneous imaging, by multiphoton microscopy, of Green Fluorescent Protein, Cyan Fluorescent Protein and collagen in vivo in living tumors. This novel method enables: 1 the simultaneous visualization of overall cell shape and sub-cellular structures such as the plasma membrane or proteins of interest in cells inside living animals, 2 direct comparison of the behavior of single cells from different cell lines in the same microenvironment in vivo. Conclusion Using this multi-fluor, multiphoton technique, we demonstrate that motility and metastatic differences between carcinoma cells of differing metastatic potential can be imaged in the same animal simultaneously at sub-cellular resolution.

  3. Ex vivo applications of multiphoton microscopy in urology

    Jain, Manu; Mukherjee, Sushmita

    2016-03-01

    Background: Routine urological surgery frequently requires rapid on-site histopathological tissue evaluation either during biopsy or intra-operative procedure. However, resected tissue needs to undergo processing, which is not only time consuming but may also create artifacts hindering real-time tissue assessment. Likewise, pathologist often relies on several ancillary methods, in addition to H&E to arrive at a definitive diagnosis. Although, helpful these techniques are tedious and time consuming and often show overlapping results. Therefore, there is a need for an imaging tool that can rapidly assess tissue in real-time at cellular level. Multiphoton microscopy (MPM) is one such technique that can generate histology-quality images from fresh and fixed tissue solely based on their intrinsic autofluorescence emission, without the need for tissue processing or staining. Design: Fresh tissue sections (neoplastic and non-neoplastic) from biopsy and surgical specimens of bladder and kidney were obtained. Unstained deparaffinized slides from biopsy of medical kidney disease and oncocytic renal neoplasms were also obtained. MPM images were acquired using with an Olympus FluoView FV1000MPE system. After imaging, fresh tissues were submitted for routine histopathology. Results: Based on the architectural and cellular details of the tissue, MPM could characterize normal components of bladder and kidney. Neoplastic tissue could be differentiated from non-neoplastic tissue and could be further classified as per histopathological convention. Some of the tumors had unique MPM signatures not otherwise seen on H&E sections. Various subtypes of glomerular lesions were identified as well as renal oncocytic neoplasms were differentiated on unstained deparaffinized slides. Conclusions: We envision MPM to become an integral part of regular diagnostic workflow for rapid assessment of tissue. MPM can be used to evaluate the adequacy of biopsies and triage tissues for ancillary studies

  4. Demonstration of structural alterations in experimental corneal infectious model using multiphoton microscopy

    Lo, Wen; Tan, Hsin-Yuan; Chang, Yuh-Ling; Sun, Yen; Lin, Sung-Jan; Jee, Shiou-Hwa; Dong, Chen-Yuan

    2007-02-01

    The aim of this study is to assess the application of multiphoton autofluorescence and second harmonic generation (SHG) microscopy for investigating the structural alterations and the pattern of microbial spreading during corneal infectious process in an in vitro organ culture model. The autofluorescence spectrum derived from pathogens allows us to monitoring the pattern of microbial spreading within corneal lamellae. In addition, the destruction and regeneration of second harmonic generating collagen during infectious process can also be monitored in a non-invasive fashion. Therefore we propose that multiphoton microscopy may potentially be applied as an effective monitoring tool for corneal infection studies.

  5. Characteristics of subgingival calculus detection by multiphoton fluorescence microscopy

    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. Imaging carious dental tissues with multiphoton fluorescence lifetime imaging microscopy

    Lin, Po-Yen; Lyu, Hong-Chou; Hsu, Chin-Ying Stephen; Chang, Chia-Seng; Kao, Fu-Jen

    2010-01-01

    In this study, multiphoton excitation was utilized to image normal and carious dental tissues noninvasively. Unique structures in dental tissues were identified using the available multimodality (second harmonic, autofluorescence, and fluorescence lifetime analysis) without labeling. The collagen in dentin exhibits a strong second harmonic response. Both dentin and enamel emit strong autofluorescence that reveals in detail morphological features (such as dentinal tubules and enamel rods) and,...

  7. Multiphoton Microscopy of Nonfluorescent Nanoparticles In Vitro and In Vivo.

    Dietzel, Steffen; Hermann, Stefanie; Kugel, Yan; Sellner, Sabine; Uhl, Bernd; Hirn, Stephanie; Krombach, Fritz; Rehberg, Markus

    2016-06-01

    Nanotechnology holds great promise for a plethora of potential applications. The interaction of engineered nanomaterials with living cells, tissues, and organisms is, however, only partly understood. Microscopic investigations of nano-bio interactions are mostly performed with a few model nanoparticles (NPs) which are easy to visualize, such as fluorescent quantum dots. Here the possibility to visualize nonfluorescent NPs with multiphoton excitation is investigated. Signals from silver (Ag), titanium dioxide (TiO2 ), and silica (SiO2 ) NPs in nonbiological environments are characterized to determine signal dependency on excitation wavelength and intensity as well as their signal stability over time. Ag NPs generate plasmon-induced luminescence decaying over time. TiO2 NPs induce photoluminescent signals of variable intensities and in addition strong third harmonic generation (THG). Optimal settings for microscopic detection are determined and then applied for visualization of these two particle types in living cells, in murine muscle tissue, and in the murine blood stream. Silica NPs produce a THG signal, but in living cells it cannot be discriminated sufficiently from endogenous cellular structures. It is concluded that multiphoton excitation is a viable option for studies of nano-bio interactions not only for fluorescent but also for some types of nonfluorescent NPs. PMID:27120195

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

    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)

  9. Nanoparticle-assisted-multiphoton microscopy for in vivo brain imaging of mice

    Qian, Jun

    2015-03-01

    Neuro/brain study has attracted much attention during past few years, and many optical methods have been utilized in order to obtain accurate and complete neural information inside the brain. Relying on simultaneous absorption of two or more near-infrared photons by a fluorophore, multiphoton microscopy can achieve deep tissue penetration and efficient light detection noninvasively, which makes it very suitable for thick-tissue and in vivo bioimaging. Nanoparticles possess many unique optical and chemical properties, such as anti-photobleaching, large multiphoton absorption cross-section, and high stability in biological environment, which facilitates their applications in long-term multiphoton microscopy as contrast agents. In this paper, we will introduce several typical nanoparticles (e.g. organic dye doped polymer nanoparticles and gold nanorods) with high multiphoton fluorescence efficiency. We further applied them in two- and three-photon in vivo functional brain imaging of mice, such as brain-microglia imaging, 3D architecture reconstruction of brain blood vessel, and blood velocity measurement.

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

    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.

  11. Large field of view multiphoton microscopy of human skin

    Balu, Mihaela; Mikami, Hideharu; Hou, Jue; Potma, Eric O.; Tromberg, Bruce J.

    2016-03-01

    Clinical examination crucially relies on the ability to quickly examine large tissue areas and rapidly zoom in to regions of interest. Skin lesions often show irregularity in color and appearance in general, especially when they start to progress towards malignancy. Large field of view (FOV) and automatic translation of the imaging area are critical in the assessment of the entire lesion. Imaging of limited FOVs of the lesion can easily result in false negative diagnosis. We present a multiphoton microscope based on two-photon excited fluorescence and second-harmonic generation that images FOVs of about 0.8 mm2 (without stitching adjacent FOVs) at speeds of 10 frames/second (800 x 800 pixels) with lateral and axial resolutions of 0.5 μm and 2.5 μm, respectively. The main novelty of this instrument is the design of the scan head, which includes a fast galvanometric scanner, relay optics, a beam expander and a high NA objective lens. We optimized the system based on the Olympus 25x, 1.05NA water immersion lens, that features a long working distance of 1 mm. Proper tailoring of the beam expander, which consists of the scan and tube lens elements, enables scaling of the FOV. The design criteria include a flat wavefront of the beam, minimum field curvature, and suppressed spherical aberrations. All aberrations in focus are below the Marechal criterion of 0.07λ rms for diffraction-limited performance. We demonstrate the practical utility of this microscope by ex-vivo imaging of wide FOVs in normal human skin.

  12. Predicting bulk mechanical properties of cellularized collagen gels using multiphoton microscopy

    Raub, CB; Putnam, AJ; Tromberg, BJ; George, SC

    2010-01-01

    Cellularized collagen gels are a common model in tissue engineering, but the relationship between the microstructure and bulk mechanical properties is only partially understood. Multiphoton microscopy (MPM) is an ideal non-invasive tool to examine collagen microstructure, cellularity and crosslink content in these gels. In order to identify robust image parameters that characterize microstructural determinants of the bulk elastic modulus, we performed serial MPM and mechanical tests on acellu...

  13. Noninvasive Assessment of Collagen Gel Microstructure and Mechanics Using Multiphoton Microscopy

    Raub, Christopher B.; Suresh, Vinod; Krasieva, Tatiana; Lyubovitsky, Julia; Mih, Justin D.; Putnam, Andrew J.; Tromberg, Bruce J.; George, Steven C.

    2006-01-01

    Multiphoton microscopy of collagen hydrogels produces second harmonic generation (SHG) and two-photon fluorescence (TPF) images, which can be used to noninvasively study gel microstructure at depth (∼1 mm). The microstructure is also a primary determinate of the mechanical properties of the gel; thus, we hypothesized that bulk optical properties (i.e., SHG and TPF) could be used to predict bulk mechanical properties of collagen hydrogels. We utilized polymerization temperature (4–37°C) and gl...

  14. Non-descanned multifocal multiphoton microscopy with a multianode photomultiplier tube

    Jae Won Cha; Yew, Elijah Y. S.; Daekeun Kim; Jaichandar Subramanian; Elly Nedivi; 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 config...

  15. Continuum generation in ultra high numerical aperture fiber with application to multiphoton microscopy

    Sayler, Nicholas

    Nonlinear microscopy benefits from broadband laser sources, enabling efficient excitation of an array of fluorophores, for example. This work demonstrates broadening of a narrow band input pulse (6 nm to 40 nm) centered at 1040 nm with excellent shot-to-shot stability. In a preliminary demonstration, multiphoton imaging with pulses from the fiber is performed. In particular second harmonic imaging of corn starch is performed.

  16. Real-time digital signal processing in multiphoton and time-resolved microscopy

    Wilson, Jesse W.; Warren, Warren S.; Fischer, Martin C.

    2016-03-01

    The use of multiphoton interactions in biological tissue for imaging contrast requires highly sensitive optical measurements. These often involve signal processing and filtering steps between the photodetector and the data acquisition device, such as photon counting and lock-in amplification. These steps can be implemented as real-time digital signal processing (DSP) elements on field-programmable gate array (FPGA) devices, an approach that affords much greater flexibility than commercial photon counting or lock-in devices. We will present progress toward developing two new FPGA-based DSP devices for multiphoton and time-resolved microscopy applications. The first is a high-speed multiharmonic lock-in amplifier for transient absorption microscopy, which is being developed for real-time analysis of the intensity-dependence of melanin, with applications in vivo and ex vivo (noninvasive histopathology of melanoma and pigmented lesions). The second device is a kHz lock-in amplifier running on a low cost (50-200) development platform. It is our hope that these FPGA-based DSP devices will enable new, high-speed, low-cost applications in multiphoton and time-resolved microscopy.

  17. Multi-photon excitation microscopy for advanced biomedical imaging

    Gadella, B.M.; van Haeften, T.W.; Bavel, Kees van; Valentijn, Jack A.

    2003-01-01

    Fluorescence microscopy (FM) is a technique traditionally used for determining biological structures [33]; its basic concept is summarised in Figure 1a. The biological specimen under examination is labelled with one or more fluorescent probes before being placed in the microscope. A single photon from the light source (usually a Hg lamp) has sufficient energy to excite an electron in the fluorescence moiety of the specimen-bound probe, taking it from an un-excited 'ground' state to an excited...

  18. Integrated structural and functional optical imaging combining spectral-domain optical coherence and multiphoton microscopy

    Vinegoni, C; Luo, W; Marks, D L; Ralston, T; Tan, W

    2005-01-01

    An integrated microscope that combines different optical techniques for simultaneous imaging is demonstrated. The microscope enables spectral-domain optical coherence microscopy based on optical backscatter, and multi-photon microscopy for the detection of two-photon fluorescence and second harmonic generation signals. The unique configuration of this integrated microscope allows for the simultaneous acquisition of both anatomical (structural) and functional imaging information with particular emphasis for applications in the fields of tissue engineering and cell biology. In addition, the contemporary analysis of the spectroscopic features can enhance contrast by differentiating among different tissue components.

  19. Intrinsic indicator of photodamage during label-free multiphoton microscopy of cells and tissues.

    Roberta Galli

    Full Text Available Multiphoton imaging has evolved as an indispensable tool in cell biology and holds prospects for clinical applications. When addressing endogenous signals such as coherent anti-Stokes Raman scattering (CARS or second harmonic generation, it requires intense laser irradiation that may cause photodamage. We report that increasing endogenous fluorescence signal upon multiphoton imaging constitutes a marker of photodamage. The effect was studied on mouse brain in vivo and ex vivo, on ex vivo human brain tissue samples, as well as on glioblastoma cells in vitro, demonstrating that this phenomenon is common to a variety of different systems, both ex vivo and in vivo. CARS microscopy and vibrational spectroscopy were used to analyze the photodamage. The development of a standard easy-to-use model that employs rehydrated cryosections allowed the characterization of the irradiation-induced fluorescence and related it to nonlinear photodamage. In conclusion, the monitoring of endogenous two-photon excited fluorescence during label-free multiphoton microscopy enables to estimate damage thresholds ex vivo as well as detect photodamage during in vivo experiments.

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

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

    2014-06-01

    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.

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

    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)

  2. In vivo multiphoton fluorescence microscopy of epithelial precancer

    Zheng, Wei; Li, Dong; Zeng, Yan; Qu, Jianan Y.

    2011-03-01

    Most human cancers arise from epithelium, the superficial layer covering the exterior of body or lining the internal body cavities. Endogenous fluorophores such as aromatic amino acids, reduced nicotinamide adenine dinucleotide (NADH), flavoprotein (FAD), keratin, collagen, and elastin can provide abundant information to reveal the changes in biochemistry, metabolism, and morphology of living tissues. Thus, autofluorescence spectroscopy and microscopy have been recognized as potential tools for discrimination of cancer from normal tissues. However, current fluorescence diagnostic studies mostly rely on spectral analysis or morphological differentiation. It is challenged since the emission spectra of endogenous fluorophores are broad and usually overlapping with each other and the fluorescence intensity could be affected by many factors. In this study, we instrumented a nonlinear optical microscopy system to characterize the morphologic and biochemical features in the epithelial precancer in vivo. The 7,12-dimethylbenz(a)anthracenetreated hamster cheek pouch were used as a living animal carcinogenesis model. And the autofluorescence signals of NADH, collagen and elastin were recorded by a time- and spectral- resolved detection system. The results show that there are obvious differences in the morphology of three-dimensional autofluorescence images between normal and precancerous epithelial tissues. The fluorescence lifetime of NADH and the SHG signal from collagen could provide additional approaches to identify cancer from normal tissue.

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

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

    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...

  4. Analysis of somitogenesis using multiphoton laser scanning microscopy (MPLSM)

    Dickinson, Mary E.; Longmuir, Kenneth J.; Fraser, Scott E.

    2001-04-01

    In order to study complex cellular interactions in the developing somite and nervous system, we have been refining techniques for labeling and imaging individual cells within the living vertebrate embryo. Most recently, we have been using MPLSM to analyze cellular behaviors, such as cell migration, filopodial extension, cell process collapse, and neuron pathfinding using time-lapse microscopy in 3-dimensions (3-d). To enhance the efficiency of two-photon excitation in these samples, we have been using a Zeiss LSM 510 NLO fiber delivery system with a Grating Dispersion Compensator (GDC). This system not only offers the convenience of fiber delivery for coupling our Ti:Sapphire laser to the microscope, but also affords us precise control over the pulsewidth of the mode- locked beam. In addition, we have developed a novel peptide/non-cationic lipid gene delivery system to introduce GFP plasmid into somite cells. This approach has allowed us to generate detailed 3-d images of somite cell morphologies at various stages of somite development in a way that best preserves the vitality of the cells being imaged.

  5. 3D super-resolved in vitro multiphoton microscopy by saturation of excitation

    Nguyen, Anh Dung; Bouwens, Arno; Vanholsbeeck, Frédérique; Egrise, Dominique; Van Simayes, Gaetan; Emplit, Philippe; Goldman, Serge; Gorza, Simon-Pierre

    2015-01-01

    We demonstrate a significant resolution enhancement beyond the conventional limit in multiphoton microscopy (MPM) using saturated excitation of fluorescence. Our technique achieves super-resolved imaging by temporally modulating the excitation laser-intensity and demodulating the higher harmonics from the saturated fluorescence signal. The improvement of the lateral and axial resolutions is measured on a sample of fluorescent microspheres. While the third harmonic already provides an enhanced resolution, we show that a further improvement can be obtained with an appropriate linear combination of the demodulated harmonics. Finally, we present in vitro imaging of fluorescent microspheres incorporated in HeLa cells to show that this technique performs well in biological samples.

  6. Novel techniques with multiphoton microscopy: Deep-brain imaging with microprisms, neurometabolism of epilepsy, and counterfeit paper money detection

    Chia, Thomas H.

    Multiphoton microscopy is a laser-scanning fluorescence imaging method with extraordinary potential. We describe three innovative multiphoton microscopy techniques across various disciplines. Traditional in vivo fluorescence microscopy of the mammalian brain has a limited penetration depth (counterfeit paper money because of its significant differences in fluorescence lifetime when compared to genuine paper money. We used scanning multiphoton laser excitation to sample a ˜4 mm2 region from 54 genuine Reserve Notes. Three types of counterfeit samples were tested. Four out of the nine counterfeit samples fit to a one-component decay. Five out of nine counterfeit samples fit to a two-component model, but are identified as counterfeit due to significant deviations in the longer lifetime component compared to genuine bills.

  7. 多焦点多光子显微技术及其研究进展%Multifocal Multiphoton Microscopy and Its Research Progress

    刘立新

    2011-01-01

    Multifocal multiphoton microscopy ( MMM) can greatly improve the utilization of the excitation light and imaging speed, and it can also acquire three-dimensional fluorescence microscopic images by multiphoton excitation with the advantages of high speed, reduced photobleaching and photodamage, enhanced penetration depth and high signal to noise ratio. The realization and research progresses of multifocal multiphoton microscopy are discussed in detail, which include simultaneous time- and spectrum-resolved multifocal multiphoton microscopy(STSR-MMM) , multifocal multiphoton microscopy based on time-correlated single photon counting( TCSPC-MMM ) , multifocal multiphoton microscopy using a field of view zoom scanning protocol and stochastic scanning multifocal multiphoton microscopy (SS-MMM) , etc.%多焦点多光子显微技术(multifocal multiphoton microscopy,MMM)提高了激发光能的利用率和成像速度,可以实现样品的三维快速多光子激发荧光显微成像,并具有对活体样品损伤小,成像深度大,图像信噪比高等优点.详细阐述了MMM的实现方法及其研究进展,包括同时时间和光谱分辨的MMM(simultaneous time- and spectrum-resolved multifocal multiphoton microscopy,STSR-MMM)、基于时间相关单光子计数技术的MMM( multifocal multiphoton microscopy based on time-correlated single photon counting,TCSPC-MMM)、基于随机扫描的MMM(stochastic scanning multifocal multiphoton microscopy,SS-MMM)、基于固定光路系统的变视场扫描的MMM等技术.

  8. Multiphoton microscopy as a diagnostic tool for pathological analysis of sentinel lymph nodes

    Lemiere, J.; Douady, J.; Estève, F.; Salameire, D.; Lantuejoul, S.; Lorimier, P.; Ricard, C.; van der Sanden, B.; Vial, J.-C.

    2009-02-01

    Multiphoton microscopy has shown a powerful potential for biomedical in vivo and ex vivo analysis of tissue sections and explants. Studies were carried out on several animal organs such as brain, arteries, lungs, and kidneys. One of the current challenges is to transfer to the clinic the knowledge and the methods previously developed in the labs at the preclinical level. For tumour staging, physicians often remove the lymph nodes that are localized at the proximity of the lesion. In case of breast cancer or melanoma, sentinel lymph node protocol is performed: pathologists randomly realize an extensive sampling of formol fixed nodes. However, the duration of this protocol is important and its reliability is not always satisfactory. The aim of our study was to determine if multiphoton microscopy would enable the fast imaging of lymph nodes on important depths, with or without exogenous staining. Experiments were first conducted on pig lymph nodes in order to test various dyes and to determine an appropriate protocol. The same experiments were then performed on thin slices of human lymph nodes bearing metastatic melanoma cells. We obtained relevant images with both endofluorescence plus second-harmonic generation and xanthene dyes. They show a good contrast between tumour and healthy cells. Furthermore, images of pig lymph nodes were recorded up to 120μm below the surface. This new method could then enable a faster diagnosis with higher efficiency for the patient. Experiments on thicker human lymph nodes are currently underway in order to validate these preliminary results.

  9. Spatiotemporal control of degenerate multiphoton fluorescence microscopy with delay-tunable femtosecond pulse pairs

    Das, Dhiman; Bhattacharyya, Indrajit; Goswami, Debabrata

    2016-07-01

    Selective excitation of a particular fluorophore in an ensemble of different fluorophores with overlapping fluorescence spectra is shown to be dependent on the time delay of femtosecond pulse pairs in multiphoton fluorescence microscopy. In particular, the two-photon fluorescence behavior of the Texas Red and DAPI dye pair inside Bovine Pulmonary Artery Endothelial (BPAE) cells depends strongly on the center wavelength of the laser, as well as the delay between two identical laser pulses in one-color femtosecond pulse-pair excitation scheme. Thus, we present a novel design concept using pairs of femtosecond pulses at different central wavelengths and tunable pulse separations for controlling the image contrast between two spatially and spectrally overlapping fluorophores. This femtosecond pulse-pair technique is unique in utilizing the variation of dye dynamics inside biological cells as a contrast mode in microscopy of different fluorophores.

  10. Label-free monitoring of colorectal adenoma-carcinoma sequence based on multiphoton microscopy

    Chen, J. X.; Li, H. S.; Chen, Z. F.; Feng, C. Y.; Yang, Y. H.; Jiang, W. Z.; Guan, G. X.; Zhu, X. Q.; Zhuo, S. M.; Xu, J.

    2014-06-01

    The monitoring and evaluation of colorectal adenoma-carcinoma sequence during endoscopy are important for endoscopic resection of precursor lesions to disrupt the adenoma-carcinoma sequence and halt progression to invasive neoplastic disease. In this study, multiphoton microscopy (MPM) was used to identify different stages during the development of colorectal adenocarcinoma including adenoma with low-grade and high-grade dysplasia, and adenocarcinoma invading the submucosa. It was found that by combining two-photon excited fluorescence (TPEF) imaging and second harmonic generation (SHG) imaging, MPM can reveal the morphological changes of the epithelial cells and glands, identify the invasive position and depth of atypical glands and quantitatively describe the change of the cellular nucleus and the nuclear-to-cytoplasmic ratio during the stepwise progression of colorectal adenocarcinoma. These are important pathological findings for pathologists when diagnosing colorectal lesions. With the advancement of a compact and flexible multiphoton endoscope for in vivo imaging and clinical applications, MPM has the potential to provide immediate histological diagnosis for the monitoring and evaluation of the colorectal adenoma-carcinoma sequence during endoscopy.

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

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

    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

  13. Use of multiphoton microscopy to diagnose liver cancer and lung metastasis in an orthotopic rat model.

    Yan, Jun; Zhuo, Shuangmu; Chen, Gang; Tan, Changjun; Zhu, Weifeng; Lu, Jianping; Fan, Jia; Chen, Jianxin; Zhou, Jian

    2012-01-01

    Liver or lung biopsy for suspicious lesions has several disadvantages such as bleeding, bile leak or pneumothorax, needle track seeding, and time-consuming histopathological procedure. The ability to directly observe cellular and subcellular details and then perform "optical biopsy" is a major goal in the development of new interventional techniques. Multiphoton microscopy (MPM) enables real-time noninvasive visualization of tissue architecture and cell morphology in live tissue. We performed a study to evaluate whether MPMcan make real-time optical diagnosis for liver cancer and lung metastasis using an orthotopic rat model with Morris hepatoma. We found that real-time high-resolution MPMimaging could clearly show tissue architecture and cell morphology. In the normal liver tissue, MPMimaging clearly revealed the blood-filled sinusoids and cords of hepatocytes. In the cancerous tissue, MPMimaging clearly illustrated that cancer cells displayed marked cellular and nuclear pleomorphism. MPMimages were comparable to golden standard hematoxylin-eosin staining images. Moreover, MPMimaging had deep penetration with the capability of optical sectioning. In short, MPMcan make real-time optical diagnosis for liver cancer and lung metastasis. This study provides the groundwork for further using multiphoton endoscopy to perform real-time noninvasive "optical biopsy" for liver cancer and lung metastasis in the near future. PMID:22331704

  14. Tunable fibre-coupled multiphoton microscopy with a negative curvature fibre.

    Sherlock, Ben; Yu, Fei; Stone, Jim; Warren, Sean; Paterson, Carl; Neil, Mark A A; French, Paul M W; Knight, Jonathan; Dunsby, Chris

    2016-07-01

    Negative curvature fibre (NCF) guides light in its core by inhibiting the coupling of core and cladding modes. In this work, an NCF was designed and fabricated to transmit ultrashort optical pulses for multiphoton microscopy with low group velocity dispersion (GVD) at 800 nm. Its attenuation was measured to be power of ∼20 mW and pulse repetition rate of 80 MHz, the NCF enabled pulses with a duration of pulse widths were maintained to within 10% of low power values up to the maximum fibre output power achievable with the laser system used of 278 mW at 700 nm, 808 mW at 800 nm and 420 mW at 860 nm. When coupled to a multiphoton microscope, it enabled imaging of ex vivo tissue using excitation wavelengths from 740 nm to 860 nm without any need for adjustments to the set-up. PMID:26989868

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

    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.

  16. Imaging normal and cancerous human gastric muscular layer in transverse and longitudinal sections by multiphoton microscopy.

    Zhou, Yi; Kang, Deyong; Yang, Zhenrong; Li, Lianhuang; Zhuo, Shuangmu; Zhu, Xiaoqin; Zhou, Yongjian; Chen, Jianxin

    2016-07-01

    Multiphoton microscopy (MPM) based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) has been widely used for imaging microstructure of biological tissues. In this article, we used MPM to investigate the microstructure changes of normal and cancerous human gastric muscular layer in transverse and longitudinal sections. The results displayed different patterns of microstructure changes of smooth muscular tissue, cell morphology and interstitial fibers in transverse and longitudinal sections, being similar to standard histopathological images but without the need for tissue processing. Our study demonstrated that MPM can bring more detailed complementary information on tissue architecture through observing transverse and longitudinal sections of tissues, which are the important pathological information when the pathologists diagnose the gastrointestinal lesions. These observations indicate that MPM could be an important potential tool to provide real-time pathological diagnosis for gastric cancer in the future. SCANNING 38:357-364, 2016. © 2015 Wiley Periodicals, Inc. PMID:26435529

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

    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)

  18. Four-dimensional multiphoton microscopy with time-correlated single-photon counting.

    Schönle, A; Glatz, M; Hell, S W

    2000-12-01

    We report on the implementation of fluorescence-lifetime imaging in multiphoton excitation microscopy that uses PC-compatible modules for time-correlated single-photon counting. Four-dimensional data stacks are produced with each pixel featuring fluorescence-decay curves that consist of as many as 4096 bins. Fluorescence lifetime(s) and their amplitude(s) are extracted by statistical methods at each pixel or in arbitrarily defined regions of interest. When employing an avalanche photodiode the width of the temporal response function is 420 ps. Although this response confines the temporal resolution to values greater than several hundreds of picoseconds, the lifetime precision is determined by the signal-to-noise ratio and can be in the range of tens of picosconds. Lifetime changes are visualized in pulsed-laser-deposited fluorescent layers as well as in cyan fluorescent proteins that transfer energy to yellow fluorescent proteins in live mammalian cells. PMID:18354639

  19. In vivo measurements of cutaneous melanin across spatial scales: Using multiphoton microscopy and spatial frequency domain spectroscopy

    Saager, RB; Balu, M; Crosignani, V; Sharif, A; Durkin, AJ; Kelly, KM; Tromberg, BJ

    2015-01-01

    © 2015 The Authors. The combined use of nonlinear optical microscopy and broadband reflectance techniques to assess melanin concentration and distribution thickness in vivo over the full range of Fitzpatrick skin types is presented. Twelve patients were measured using multiphoton microscopy (MPM) and spatial frequency domain spectroscopy (SFDS) on both dorsal forearm and volar arm, which are generally sun-exposed and non-sun-exposed areas, respectively. Both MPM and SFDS measured melanin volu...

  20. Investigation of signal-to-noise ratio in frequency-domain multiphoton fluorescence lifetime imaging microscopy.

    Zhang, Yide; Khan, Aamir A; Vigil, Genevieve D; Howard, Scott S

    2016-07-01

    Multiphoton microscopy (MPM) combined with fluorescence lifetime imaging microscopy (FLIM) has enabled three-dimensional quantitative molecular microscopy in vivo. The signal-to-noise ratio (SNR), and thus the imaging rate of MPM-FLIM, which is fundamentally limited by the shot noise and fluorescence saturation, has not been quantitatively studied yet. In this paper, we investigate the SNR performance of the frequency-domain (FD) MPM-FLIM with two figures of merit: the photon economy in the limit of shot noise, and the normalized SNR in the limit of saturation. The theoretical results and Monte Carlo simulations find that two-photon FD-FLIM requires 50% fewer photons to achieve the same SNR as conventional one-photon FLIM. We also analytically show that the MPM-FD-FLIM can exploit the DC and higher harmonic components generated by nonlinear optical mixing of the excitation light to improve SNR, reducing the required number of photons by an additional 50%. Finally, the effect of fluorophore saturation on the experimental SNR performance is discussed. PMID:27409702

  1. Label-free in vivo imaging of Drosophila melanogaster by multiphoton microscopy

    Lin, Chiao-Ying; Hovhannisyan, Vladimir; Wu, June-Tai; Lin, Sung-Jan; Lin, Chii-Wann; Chen, Jyh-Horng; Dong, Chen-Yuan

    2008-02-01

    The fruit fly Drosophila melanogaster is one of the most valuable organisms in genetic and developmental biology studies. Drosophila is a small organism with a short life cycle, and is inexpensive and easy to maintain. The entire genome of Drosophila has recently been sequenced (cite the reference). These advantages make fruit fly an attractive model organism for biomedical researches. Unlike humans, Drosophila can be subjected to genetic manipulation with relative ease. Originally, Drosophila was mostly used in classical genetics studies. In the model era of molecular biology, the fruit fly has become a model organ for developmental biology researches. In the past, numerous molecularly modified mutants with well defined genetic defects affecting different aspects of the developmental processes have been identified and studied. However, traditionally, the developmental defects of the mutant flies are mostly examined in isolated fixed tissues which preclude the observation of the dynamic interaction of the different cell types and the extracellular matrix. Therefore, the ability to image different organelles of the fruit fly without extrinsic labeling is invaluable for Drosophila biology. In this work, we successfully acquire in vivo images of both developing muscles and axons of motor neurons in the three larval stages by using the minimially invasive imaging modality of multiphoton (SHG) microscopy. We found that while SHG imaging is useful in revealing the muscular architecture of the developing larva, it is the autofluorescence signal that allows label-free imaging of various organelles to be achieved. Our results demonstrate that multiphoton imaging is a powerful technique for investigation the development of Drosophila.

  2. Multimodal microscopy and the stepwise multi-photon activation fluorescence of melanin

    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

  3. Simultaneous multiple-excitation multiphoton microscopy yields increased imaging sensitivity and specificity

    Brinkman Brendan C

    2011-03-01

    Full Text Available Abstract Background Multiphoton microscopy (MPM offers many advantages over conventional wide-field and confocal laser scanning microscopy (CLSM for imaging biological samples such as 3D resolution of excitation, reduced phototoxicity, and deeper tissue imaging. However, adapting MPM for critical multi-color measurements presents a challenge because of the largely overlapping two-photon absorption (TPA peaks of common biological fluorophores. Currently, most multi-color MPM relies on the absorbance at one intermediate wavelength of multiple dyes, which introduces problems such as decreased and unequal excitation efficiency across the set of dyes. Results Here we describe an MPM system incorporating two, independently controlled sources of two-photon excitation whose wavelengths are adjusted to maximally excite one dye while minimally exciting the other. We report increased signal-to-noise ratios and decreased false positive emission bleed-through using this novel multiple-excitation MPM (ME-MPM compared to conventional single-excitation MPM (SE-MPM in a variety of multi-color imaging applications. Conclusions Similar to the tremendous gain in popularity of CLSM after the introduction of multi-color imaging, we anticipate that the ME-MPM system will further increase the popularity of MPM. In addition, ME-MPM provides an excellent tool to more rapidly design and optimize pairs of fluorescence probes for multi-color two-photon imaging, such as CFP/YFP or GFP/DsRed for CLSM.

  4. Super-resolved multimodal multiphoton microscopy with spatial frequency-modulated imaging

    Field, Jeffrey J; Domingue, Scott R; Motz, Alyssa M Allende; DeLuca, Keith F; DeLuca, Jennifer G; Kuciauskas, Darius; Levi, Dean H; Squier, Jeff A; Bartels, Randy A

    2015-01-01

    Super-resolved far-field microscopy has emerged as a powerful tool for investigating the structure of objects with resolution well below the diffraction limit of light. Nearly all super-resolution imaging techniques reported to date rely on real energy states of probe molecules to circumvent the diffraction limit, preventing super-resolved imaging of contrast mechanisms that occur via virtual energy states such as harmonic generation (HG). Here we report a super-resolution technique based on SPatIal Frequency modulated Imaging (SPIFI) that permits super-resolved nonlinear microscopy with any contrast mechanism, and with single-pixel detection. We show multimodal super-resolved images with two-photon excited fluorescence (TPEF) and second-harmonic generation (SHG) from biological and inorganic media. Multiphoton SPIFI (MP-SPIFI) provides spatial resolution up to 2$\\eta$ below the diffraction limit, where $\\eta$ is the highest power of the nonlinear intensity response. MP-SPIFI has the potential to not only pro...

  5. Multiphoton microscopy, fluorescence lifetime imaging and optical spectroscopy for the diagnosis of neoplasia

    Skala, Melissa Caroline

    2007-12-01

    Cancer morbidity and mortality is greatly reduced when the disease is diagnosed and treated early in its development. Tissue biopsies are the gold standard for cancer diagnosis, and an accurate diagnosis requires a biopsy from the malignant portion of an organ. Light, guided through a fiber optic probe, could be used to inspect regions of interest and provide real-time feedback to determine the optimal tissue site for biopsy. This approach could increase the diagnostic accuracy of current biopsy procedures. The studies in this thesis have characterized changes in tissue optical signals with carcinogenesis, increasing our understanding of the sensitivity of optical techniques for cancer detection. All in vivo studies were conducted on the dimethylbenz[alpha]anthracene treated hamster cheek pouch model of epithelial carcinogenesis. Multiphoton microscopy studies in the near infrared wavelength region quantified changes in tissue morphology and fluorescence with carcinogenesis in vivo. Statistically significant morphological changes with precancer included increased epithelial thickness, loss of stratification in the epithelium, and increased nuclear diameter. Fluorescence changes included a statistically significant decrease in the epithelial fluorescence intensity per voxel at 780 nm excitation, a decrease in the fluorescence lifetime of protein-bound nicotinamide adenine dinucleotide (NADH, an electron donor in oxidative phosphorylation), and an increase in the fluorescence lifetime of protein-bound flavin adenine dinucleotide (FAD, an electron acceptor in oxidative phosphorylation) with precancer. The redox ratio (fluorescence intensity of FAD/NADH, a measure of the cellular oxidation-reduction state) did not significantly change with precancer. Cell culture experiments (MCF10A cells) indicated that the decrease in protein-bound NADH with precancer could be due to increased levels of glycolysis. Point measurements of diffuse reflectance and fluorescence spectra in

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

    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.

  7. Label-free identification of the hippocampus and surrounding structures by multiphoton microscopy

    Wang, Shu; Jiang, Liwei; Du, Huiping; Wang, Xingfu; Zheng, Liqin; Li, Lianhuang; Zhuo, Shuangmu; Zhu, Xiaoqin; Chen, Jianxin

    2016-05-01

    The hippocampus is one of the essential neuroanatomical substrates and plays an important role in different neurological illnesses. In this work, multiphoton microscopy (MPM) based on intrinsic nonlinear optical processes two-photon excited fluorescence (TPEF) and second harmonic generation (SHG), was applied to label-freely detect the entire hippocampus and surrounding structures in high-magnification imaging, as well as acquire large-scale MPM images at subcellular resolution. It was found that MPM has the capability to identify cornu ammonis, dentate gyrus (DG), alveus, and fimbria of the entire hippocampus, choroid plexus in lateral ventricles, and white matter tracts. MPM also can be used to quantitatively describe the differences of the cellular nucleus in the cornu ammonis and the DG, further identify the morphological features of hippocampal subfields. In addition, the surrounding structures of the hippocampus including the lateral ventricles and white matter serve as useful information to determine the position of the hippocampus. Our results suggest that with the development of the clinical feasibility of two-photon fiberscopes and microendoscope probes, MPM has the potential for in vivo intraoperative identification and monitoring of hippocampus-related lesions without the need for tissue labelling or fluorescent markers.

  8. Non-descanned multifocal multiphoton microscopy with a multianode photomultiplier tube

    Jae Won Cha

    2015-08-01

    Full Text Available 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.

  9. Digital deconvolution filter derived from linear discriminant analysis and application for multiphoton fluorescence microscopy.

    Sullivan, Shane Z; Schmitt, Paul D; Muir, Ryan D; DeWalt, Emma L; Simpson, Garth J

    2014-04-01

    A digital filter derived from linear discriminant analysis (LDA) is developed for recovering impulse responses in photon counting from a high speed photodetector (rise time of ~1 ns) and applied to remove ringing distortions from impedance mismatch in multiphoton fluorescence microscopy. Training of the digital filter was achieved by defining temporally coincident and noncoincident transients and identifying the projection within filter-space that best separated the two classes. Once trained, data analysis by digital filtering can be performed quickly. Assessment of the reliability of the approach was performed through comparisons of simulated voltage transients, in which the ground truth results were known a priori. The LDA filter was also found to recover deconvolved impulses for single photon counting from highly distorted ringing waveforms from an impedance mismatched photomultiplier tube. The LDA filter was successful in removing these ringing distortions from two-photon excited fluorescence micrographs and through data simulations was found to extend the dynamic range of photon counting by approximately 3 orders of magnitude through minimization of detector paralysis. PMID:24559143

  10. Identifying and quantifying the stromal fibrosis in muscularis propria of colorectal carcinoma by multiphoton microscopy

    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. (letter)

  11. Approach to quantify human dermal skin aging using multiphoton laser scanning microscopy

    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.

  12. Identifying three different architectural subtypes of mammary ductal carcinoma in situ using multiphoton microscopy

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

    2015-10-01

    Ductal carcinoma in situ (DCIS) is often considered as the precursor of invasive breast cancer, and the risk of DCIS progression to IBC has been estimated based on the evaluation of pathological features, among which the architectural subtype is the most common one. In this study, multiphoton microscopy (MPM) is applied to identify three different architectural subtypes of DCIS (solid, cribriform and comedo). It is found that MPM has the capability to visualize the proliferating pattern of tumor cells, the presence of intraluminal necrosis and the morphology of basement membrane, which are all taken into account in subtyping DCIS. In addition, MPM also can be used to quantify the cellular metabolism, for quantitatively identifying tumor staging during tumor progression. This result highlights the potential of MPM as an advanced technique to assess the pathological characters of the breast tumor in real-time and reflect the degree of tumor progression in vivo, by integrating into the intra-fiberoptic ductoscopy or transdermal biopsy needle.

  13. Multiphoton microscopy as a diagnostic imaging modality for pancreatic neoplasms without hematoxylin and eosin stains

    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.

  14. Fringe-free, Background-free, Collinear Third Harmonic Generation FROG Measurements for Multiphoton Microscopy

    Chadwick, R; Spahr, E; Squier, J A; Durfee, C G; Walker, B C; Fittinghoff, D N

    2006-07-21

    Collinear pulse measurement tools useful at the full numerical aperture (NA) of multiphoton microscope objectives are a necessity for a quantitative characterization of the femtosecond pulses focused by these systems. In this letter, we demonstrate a simple new technique, for characterizing the pulse at the focus in a multiphoton microscope. This technique, a background-free, fringe-free, form of frequency-resolved optical gating, uses the third harmonic signal generated from a glass coverslip. Here it is used to characterize 100 fs pulses (typical values for a multiphoton microscope) at the focus of a 0.65 NA objective.

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

    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.

  16. Multiphoton Microscopy in the Study of Morphological Characteristics of Radiation-Induced Injuries of the Bladder

    S.S. Kuznetsov

    2016-06-01

    Full Text Available The aim of the investigation was to assess the feasibility of multiphoton microscopy (MPM for studying dynamics of bladder structural changes following a single exposure to gamma-radiation at various doses (2, 10, and 40 Gy in experiment. Materials and Methods. Specimens of rat bladders after a single local radiation at the dose of 2, 10, and 40 Gy were the objects of investigation (9 groups with two rats for each dose and term, and two intact rats — 20 observations in all. The study was carried out 1 day, 1 week, and 1 month after radiation exposure. Part of the histological bladder preparations was stained with picrofuchsin according to Van Gieson method. The other part of the sections, obtained from the same blocks, was investigated using MPM without additional staining. For this purpose a laser scanning microscope LSM Axiovert 510 Meta (Carl Zeiss, Germany was used. Excitation was generated with a femtosecond Ti:Sapphire laser (MAI TAI HP, Spectra Physics, USA at the wavelength of 800 nm, registration was performed in the range of 362–415 nm (second harmonic signal from collagen and 512–576 nm (signal of two-photon excited elastin autofluorescence. Results. Application of MPM method allowed us to find out, that in early terms (1 day and 1 week after radiation exposure the process of alteration of collagen-containing structures of bladder walls was a leading one at all selected doses. A month after 2 and 10 Gy radiation increase in collagen structures was registered, speaking of the onset of radiation fibrosis formation. At a dose of 40 Gy decrease of second harmonic signal retained in the extracellular matrix of the bladder wall. It allowed us to draw a conclusion on a long-term disorganization of collagen at high radiation doses. Conclusion. MPM method makes it possible to estimate, that structural destruction of extracellular tissue matrix occurs even after low radiation doses and in early terms after radiation exposure, which is not

  17. Miniature fiber-optic multiphoton microscopy system using frequency-doubled femtosecond Er-doped fiber laser

    Huang, Lin; Mills, Arthur K; Zhao, Yuan; Jones, David J.; Tang, Shuo

    2016-01-01

    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 78...

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

    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...

  19. Comparison of in vivo and ex vivo laser scanning microscopy and multiphoton tomography application for human and porcine skin imaging

    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)

  20. Comparison of in vivo and ex vivo laser scanning microscopy and multiphoton tomography application for human and porcine skin imaging

    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)

  1. Miniature fiber-optic multiphoton microscopy system using frequency-doubled femtosecond Er-doped fiber laser.

    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. PMID:27231633

  2. In vivo, label-free, three-dimensional quantitative imaging of liver surface using multi-photon microscopy

    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.

  3. In vivo three-dimensional optical coherence tomography and multiphoton microscopy in a mouse model of ovarian neoplasia

    Watson, Jennifer M.; Marion, Samuel L.; Rice, Photini Faith; Bentley, David L.; Besselsen, David; Utzinger, Urs; Hoyer, Patricia B.; Barton, Jennifer K.

    2013-03-01

    Our goal is to use optical coherence tomography (OCT) and multiphoton microscopy (MPM) to detect early tumor development in a mouse model of ovarian neoplasia. We hope to use information regarding early tumor development to create a diagnostic test for high-risk patients. In this study we collect in vivo images using OCT, second harmonic generation and two-photon excited fluorescence from non-vinylcyclohexene diepoxide (VCD)-dosed and VCD-dosed mice. VCD causes follicular apoptosis (simulating menopause) and leads to tumor development. Using OCT and MPM we visualized the ovarian microstructure and were able to see differences between non-VCD-dosed and VCD-dosed animals. This leads us to believe that OCT and MPM may be useful for detecting changes due to early tumor development.

  4. Compact non-contact total emission detection for in vivo multiphoton excitation microscopy.

    Combs, C A; Smirnov, A; Glancy, B; Karamzadeh, N S; Gandjbakhche, A H; Redford, G; Kilborn, K; Knutson, J R; Balaban, R S

    2014-02-01

    We describe a compact, non-contact design for a total emission detection (c-TED) system for intra-vital multiphoton imaging. To conform to a standard upright two-photon microscope design, this system uses a parabolic mirror surrounding a standard microscope objective in concert with an optical path that does not interfere with normal microscope operation. The non-contact design of this device allows for maximal light collection without disrupting the physiology of the specimen being examined. Tests were conducted on exposed tissues in live animals to examine the emission collection enhancement of the c-TED device compared to heavily optimized objective-based emission collection. The best light collection enhancement was seen from murine fat (5×-2× gains as a function of depth), whereas murine skeletal muscle and rat kidney showed gains of over two and just under twofold near the surface, respectively. Gains decreased with imaging depth (particularly in the kidney). Zebrafish imaging on a reflective substrate showed close to a twofold gain throughout the entire volume of an intact embryo (approximately 150 μm deep). Direct measurement of bleaching rates confirmed that the lower laser powers, enabled by greater light collection efficiency, yielded reduced photobleaching in vivo. The potential benefits of increased light collection in terms of speed of imaging and reduced photo-damage, as well as the applicability of this device to other multiphoton imaging methods is discussed. PMID:24251437

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

    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 electr...

  6. Smart microscope: an adaptive optics learning system for aberration correction in multiphoton confocal microscopy.

    Albert, O; Sherman, L; Mourou, G; Norris, T B; Vdovin, G

    2000-01-01

    Off-axis aberrations in a beam-scanning multiphoton confocal microscope are corrected with a deformable mirror. The optimal mirror shape for each pixel is determined by a genetic learning algorithm, in which the second-harmonic or two-photon fluorescence signal from a reference sample is maximized. The speed of the convergence is improved by use of a Zernike polynomial basis for the deformable mirror shape. This adaptive optical correction scheme is implemented in an all-reflective system by use of extremely short (10-fs) optical pulses, and it is shown that the scanning area of an f:1 off-axis parabola can be increased by nine times with this technique. PMID:18059779

  7. In vivo measurements of cutaneous melanin across spatial scales: using multiphoton microscopy and spatial frequency domain spectroscopy

    Saager, Rolf B.; Balu, Mihaela; Crosignani, Viera; Sharif, Ata; Durkin, Anthony J.; Kelly, Kristen M.; Tromberg, Bruce J.

    2015-06-01

    The combined use of nonlinear optical microscopy and broadband reflectance techniques to assess melanin concentration and distribution thickness in vivo over the full range of Fitzpatrick skin types is presented. Twelve patients were measured using multiphoton microscopy (MPM) and spatial frequency domain spectroscopy (SFDS) on both dorsal forearm and volar arm, which are generally sun-exposed and non-sun-exposed areas, respectively. Both MPM and SFDS measured melanin volume fractions between ˜5% (skin type I non-sun-exposed) and 20% (skin type VI sun exposed). MPM measured epidermal (anatomical) thickness values ˜30-65 μm, while SFDS measured melanin distribution thickness based on diffuse optical path length. There was a strong correlation between melanin concentration and melanin distribution (epidermal) thickness measurements obtained using the two techniques. While SFDS does not have the ability to match the spatial resolution of MPM, this study demonstrates that melanin content as quantified using SFDS is linearly correlated with epidermal melanin as measured using MPM (R2=0.8895). SFDS melanin distribution thickness is correlated to MPM values (R2=0.8131). These techniques can be used individually and/or in combination to advance our understanding and guide therapies for pigmentation-related conditions as well as light-based treatments across a full range of skin types.

  8. Correlating intravital multi-photon microscopy to 3D electron microscopy of invading tumor cells using anatomical reference points.

    Matthia A Karreman

    Full Text Available 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.

  9. Ex vivo multiscale quantitation of skin biomechanics in wild-type and genetically-modified mice using multiphoton microscopy

    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.

  10. Three-Dimensional Morphology by Multiphoton Microscopy with Clearing in a Model of Cisplatin-Induced CKD.

    Torres, Richard; Velazquez, Heino; Chang, John J; Levene, Michael J; Moeckel, Gilbert; Desir, Gary V; Safirstein, Robert

    2016-04-01

    Traditional histologic methods are limited in their ability to detect pathologic changes of CKD, of which cisplatin therapy is an important cause. In addition, poor reproducibility of available methods has limited analysis of the role of fibrosis in CKD. Highly labor-intensive serial sectioning studies have demonstrated that three-dimensional perspective can reveal useful morphologic information on cisplatin-induced CKD. By applying the new technique of multiphoton microscopy (MPM) with clearing to a new mouse model of cisplatin-induced CKD, we obtained detailed morphologic and collagen reconstructions of millimeter-thick renal sections that provided new insights into pathophysiology. Quantitative analysis revealed that a major long-term cisplatin effect is reduction in the number of cuboidal cells of the glomerular capsule, a change we term the "uncapped glomerulus lesion." Glomerulotubular disconnection was confirmed, but connection remnants between damaged tubules and atubular glomeruli were observed. Reductions in normal glomerular capsules corresponded to reductions in GFR. Mild increases in collagen were noted, but the fibrosis was not spatially correlated with atubular glomeruli. Glomerular volume and number remained unaltered with cisplatin exposure, but cortical tubulointerstitial mass decreased. In conclusion, new observations were made possible by using clearing MPM, demonstrating the utility of this technique for studies of renal disease. This technique should prove valuable for further characterizing the evolution of CKD with cisplatin therapy and of other conditions. PMID:26303068

  11. Multiphoton microscopy study of the morphological and quantity changes of collagen and elastic fiber components in keloid disease

    Chen, Jianxin; Zhuo, Shuangmu; Jiang, Xingshan; Zhu, Xiaoqin; Zheng, Liqin; Xie, Shusen; Lin, Bifang; Zeng, Haishan

    2011-05-01

    Multiphoton microscopy was used to study the extracellular matrix of keloid at the molecular level without tissue fixation and staining. Direct imaging of collagen and elastin was achieved by second harmonic generation and two-photon excited fluorescence, respectively. The morphology and quantity of collagen and elastin in keloid were characterized and quantitatively analyzed in comparison to normal skin. The study demonstrated that in keloid, collagen content increased in both the upper dermis and the deep dermis, while elastin mostly showed up in the deep dermis and its quantity is higher compared to normal skin. This suggests the possibility that abnormal fibroblasts synthesized an excessive amount of collagen and elastin at the beginning of keloid formation, corresponding to the observed deep dermis, while after a certain time point, the abnormal fibroblast produced mostly collagen, corresponding to the observed upper dermis. The morphology of collagen and elastin in keloid was disrupted and presented different variations. In the deep dermis, elastic fibers showed node structure, while collagen showed obviously regular gaps between adjacent bundles. In the upper dermis, collagen bundles aligned in a preferred direction, while elastin showed as sparse irregular granules. This new molecular information provided fresh insight about the development process of keloid.

  12. Quantifying local heterogeneity of in vivo transport dynamics using stochastic scanning multiphoton multifocal microscopy and segmented spatiotemporal image correlation spectroscopy

    Kim, Hee Y.; Jureller, Justin E.; Kuznetsov, Andrey; Philipson, Louis H.; Scherer, Norbert F.

    2008-02-01

    Elucidating the mechanisms of insulin granule trafficking in pancreatic β-cells is a critical step in understanding Type II Diabetes and abnormal insulin secretion. In this paper, rapid-sampling stochastic scanning multiphoton multifocal microscopy (SS-MMM) was developed to capture fast insulin granule dynamics in vivo. Stochastic scanning of (a diffractive optic generated) 10×10 hexagonal array of foci with a galvanometer yields a uniformly sampled image with fewer spatio-temporal artifacts than obtained by conventional or multibeam raster scanning. In addition, segmented spatio-temporal image correlation spectroscopy (Segmented STICS) was developed to extract dynamics of insulin granules from the image sequences. Measurements we conducted on MIN6 cells, which exhibit an order of magnitude lower granule number density, allow comparison of particle tracking with Segmented-STICS. Segmentation of the images into 8×8 pixel segments (similar to a size of one granule) allows some amount of spatial averaging, which can reduce the computation time required to calculate the correlation function, yet retains information about the local spatial heterogeneity of transport. This allows the correlation analysis to quantify the dynamics within each of the segments producing a "map" of the localized properties of the cell. The results obtained from Segmented STICS are compared with dynamics determined from particle tracking analysis of the same images. The resulting range of diffusion coefficients of insulin granules are comparable to previously published values indicating that SS-MMM and segmented- STICS will be useful to address the imaging challenges presented by β-cells, particularly the extremely large number density of granules.

  13. Examination of diagnostic features in multiphoton microscopy and optical coherence tomography images of ovarian tumorigenesis in a mouse model

    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

  14. Age-related changes in murine bladder structure and sensory innervation: a multiphoton microscopy quantitative analysis.

    Schueth, Anna; Spronck, Bart; van Zandvoort, Marc A M J; van Koeveringe, Gommert A

    2016-02-01

    Our study aimed to examine and quantify age-related structural alterations in the healthy mouse bladder using ex vivo two-photon laser scanning microscopy (TPLSM). Freshly dissected bladders from 25-, 52-, and 85-week-old C57bl/6J mice were examined, and morphological analyses and quantification of cell layers and nerves were performed. The numbers of stretched, curled, branched, and total number of nerves in volume units of the stained muscle layer were quantified. We observed differences in the bladder wall architecture and innervation with age. Especially in 85-week-old mice, age-related changes were found, including detachment of urothelial cells and an increase in connective tissue, intermingled with the smooth muscle fibers in the muscle layer (collagen-smooth muscle ratio of 1.15 ± 0.29). In 25- and 52-week-old mice, the collagen-smooth muscle ratios were 0.20 ± 0.04 and 0.31 ± 0.11, respectively, and a clear separation of collagen and muscle was observed. The overall number of nerves and the number of curled nerves were significantly higher in the 85-week-old mice (74.0 ± 13.0 and 25.9 ± 4.8, respectively), when comparing to 25-week-old mice (26.0 ± 2.7 and 6.7 ± 1.2, respectively) and 52-week-old mice (43.8 ± 4.3 and 22.1 ± 3.3, respectively). Significant age-related alterations in bladder morphology and innervation were found, when comparing freshly dissected bladder tissue from 25-, 52-, and 85-week-old mice. The higher number of curled nerves might be an indication of an increased neurotransmitter release, resulting in a higher nerve activity, with a part of the nerves being possibly mechanically impaired. This study shows that two-photon laser scanning microscopy of healthy aging male mice is a useful method to investigate and quantify the age-related changes in the bladder wall. PMID:26825637

  15. Functional cardiac imaging by random access microscopy

    Claudia eCrocini

    2014-10-01

    Full Text Available Advances in the development of voltage sensitive dyes and Ca2+ sensors in combination with innovative microscopy techniques allowed researchers to perform functional measurements with an unprecedented spatial and temporal resolution. At the moment, one of the shortcomings of available technologies is their incapability of imaging multiple fast phenomena while controlling the biological determinants involved. In the near future, ultrafast deflectors can be used to rapidly scan laser beams across the sample, performing optical measurements of action potential and Ca2+ release from multiple sites within cardiac cells and tissues. The same scanning modality could also be used to control local Ca2+ release and membrane electrical activity by activation of caged compounds and light-gated ion channels. With this approach, local Ca2+ or voltage perturbations could be induced, simulating arrhythmogenic events, and their impact on physiological cell activity could be explored. The development of this optical methodology will provide fundamental insights in cardiac disease, boosting new therapeutic strategies, and, more generally, it will represent a new approach for the investigation of the physiology of excitable cells.

  16. Remote z-scanning with a macroscopic voice coil motor for fast 3D multiphoton laser scanning microscopy.

    Rupprecht, Peter; Prendergast, Andrew; Wyart, Claire; Friedrich, Rainer W

    2016-05-01

    There is a high demand for 3D multiphoton imaging in neuroscience and other fields but scanning in axial direction presents technical challenges. We developed a focusing technique based on a remote movable mirror that is conjugate to the specimen plane and translated by a voice coil motor. We constructed cost-effective z-scanning modules from off-the-shelf components that can be mounted onto standard multiphoton laser scanning microscopes to extend scan patterns from 2D to 3D. Systems were designed for large objectives and provide high resolution, high speed and a large z-scan range (>300 μm). We used these systems for 3D multiphoton calcium imaging in the adult zebrafish brain and measured odor-evoked activity patterns across >1500 neurons with single-neuron resolution and high signal-to-noise ratio. PMID:27231612

  17. Quantitative structural markers of colorectal dysplasia in a cross sectional study of ex vivo murine tissue using label-free multiphoton microscopy

    Prieto, Sandra P.; Greening, Gage J.; Lai, Keith K.; Muldoon, Timothy J.

    2016-03-01

    Two-photon excitation of label-free tissue is of increasing interest, as advances have been made in endoscopic clinical application of multiphoton microscopy, such as second harmonic generation (SHG) scanning endoscopy used to monitor cervical collagen in mice1. We used C57BL mice as a model to investigate the progression of gastrointestinal structures, specifically glandular area and circularity. We used multiphoton microscopy to image ex-vivo label-free murine colon, focusing on the collagen structure changes over time, in mice ranging from 10 to 20 weeks of age. Series of images were acquired within the colonic and intestinal tissue at depth intervals of 20 microns from muscularis to the epithelium, up to a maximum depth of 180 microns. The imaging system comprised a two-photon laser tuned to 800nm wavelength excitation, and the SHG emission was filtered with a 400/40 bandpass filter before reaching the photomultiplier tube. Images were acquired at 15 frames per second, for 200 to 300 cumulative frames, with a field of view of 261um by 261um, and 40mW at sample. Image series were compared to histopathology H&E slides taken from adjacent locations. Quantitative metrics for determining differences between murine glandular structures were applied, specifically glandular area and circularity.

  18. Flexible digital signal processing architecture for narrowband and spread-spectrum lock-in detection in multiphoton microscopy and time-resolved spectroscopy

    Wilson, Jesse W.; Park, Jong Kang; Warren, Warren S.

    2015-01-01

    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. PMID:25832238

  19. Flexible digital signal processing architecture for narrowband and spread-spectrum lock-in detection in multiphoton microscopy and time-resolved spectroscopy.

    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. PMID:25832238

  20. Mitigating phototoxicity during multiphoton microscopy of live Drosophila embryos in the 1.0-1.2 µm wavelength range.

    Delphine Débarre

    Full Text Available Light-induced toxicity is a fundamental bottleneck in microscopic imaging of live embryos. In this article, after a review of photodamage mechanisms in cells and tissues, we assess photo-perturbation under illumination conditions relevant for point-scanning multiphoton imaging of live Drosophila embryos. We use third-harmonic generation (THG imaging of developmental processes in embryos excited by pulsed near-infrared light in the 1.0-1.2 µm range. We study the influence of imaging rate, wavelength, and pulse duration on the short-term and long-term perturbation of development and define criteria for safe imaging. We show that under illumination conditions typical for multiphoton imaging, photodamage in this system arises through 2- and/or 3-photon absorption processes and in a cumulative manner. Based on this analysis, we derive general guidelines for improving the signal-to-damage ratio in two-photon (2PEF/SHG or THG imaging by adjusting the pulse duration and/or the imaging rate. Finally, we report label-free time-lapse 3D THG imaging of gastrulating Drosophila embryos with sampling appropriate for the visualisation of morphogenetic movements in wild-type and mutant embryos, and long-term multiharmonic (THG-SHG imaging of development until hatching.

  1. Multiphoton microscopy based cryo-imaging of inflated frozen human lung sections at -60°C in healthy and COPD lungs

    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

  2. Deep insights: intravital imaging with two-photon microscopy.

    Schießl, Ina Maria; Castrop, Hayo

    2016-09-01

    Intravital multiphoton microscopy is widely used to assess the structure and function of organs in live animals. Although different tissues vary in their accessibility for intravital multiphoton imaging, considerable progress has been made in the imaging quality of all tissues due to substantial technical improvements in the relevant imaging components, such as optics, excitation laser, detectors, and signal analysis software. In this review, we provide an overview of the technical background of intravital multiphoton microscopy. Then, we note a few seminal findings that were made through the use of multiphoton microscopy. Finally, we address the technical limitations of the method and provide an outlook for how these limitations may be overcome through future technical developments. PMID:27352273

  3. Measurement of absorption spectrum of deuterium oxide (D2O) and its application to signal enhancement in multiphoton microscopy at the 1700-nm window

    1700-nm window has been demonstrated to be a promising excitation window for deep-tissue multiphoton microscopy (MPM). Long working-distance water immersion objective lenses are typically used for deep-tissue imaging. However, absorption due to immersion water at 1700 nm is still high and leads to dramatic decrease in signals. In this paper, we demonstrate measurement of absorption spectrum of deuterium oxide (D2O) from 1200 nm to 2600 nm, covering the three low water-absorption windows potentially applicable for deep-tissue imaging (1300 nm, 1700 nm, and 2200 nm). We apply this measured result to signal enhancement in MPM at the 1700-nm window. Compared with water immersion, D2O immersion enhances signal levels in second-harmonic generation imaging, 3-photon fluorescence imaging, and third-harmonic generation imaging by 8.1, 24.8, and 24.7 times with 1662-nm excitation, in good agreement with theoretical calculation based on our absorption measurement. This suggests D2O a promising immersion medium for deep-tissue imaging

  4. Measurement of absorption spectrum of deuterium oxide (D{sub 2}O) and its application to signal enhancement in multiphoton microscopy at the 1700-nm window

    Wang, Yuxin; Wen, Wenhui; Wang, Kai; Wang, Ke, E-mail: kewangfs@szu.edu.cn [Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Optoelectronic Engineering, Shenzhen University, Shenzhen 518060 (China); Zhai, Peng [Key Laboratory of Biomedical Engineering of Shenzhen, College of Medicine, Shenzhen University, Shenzhen 518060 (China); Qiu, Ping, E-mail: pingqiu@szu.edu.cn [College of Physics Science and Technology, Shenzhen University, Shenzhen 518060 (China)

    2016-01-11

    1700-nm window has been demonstrated to be a promising excitation window for deep-tissue multiphoton microscopy (MPM). Long working-distance water immersion objective lenses are typically used for deep-tissue imaging. However, absorption due to immersion water at 1700 nm is still high and leads to dramatic decrease in signals. In this paper, we demonstrate measurement of absorption spectrum of deuterium oxide (D{sub 2}O) from 1200 nm to 2600 nm, covering the three low water-absorption windows potentially applicable for deep-tissue imaging (1300 nm, 1700 nm, and 2200 nm). We apply this measured result to signal enhancement in MPM at the 1700-nm window. Compared with water immersion, D{sub 2}O immersion enhances signal levels in second-harmonic generation imaging, 3-photon fluorescence imaging, and third-harmonic generation imaging by 8.1, 24.8, and 24.7 times with 1662-nm excitation, in good agreement with theoretical calculation based on our absorption measurement. This suggests D{sub 2}O a promising immersion medium for deep-tissue imaging.

  5. Integrated coherent Raman scattering and multiphoton microscopy for label-free imaging of the dentin in the tooth

    Wang, Zi; Zheng, Wei; Lin, Jian; Hsu, Chin-Ying; Huang, Zhiwei

    2014-02-01

    We report the implementation of a unique multimodal nonlinear optical microscopy (i.e., coherent anti-Stokes Raman scattering (CARS), second harmonic generation (SHG), third harmonic generation (THG) and two photon excitation fluorescence (TPEF)) platform for label-free imaging of dentin. A picosecond tunable laser together with an OPO is used as the excitation source for simultaneously multimodal imaging. CARS shows similar information as TPEF in dentin, but it has a higher sectioning performance than TPEF and thus it is a good alternative for TPEF. Microtubule structure is revealed nearby dentin enamel junction (DEJ) from the multimodal images. This work demonstrates that combining different nonlinear optical imaging modalities can provide new insights into the understanding of morphological structures and biochemical/biomolecular distributions of the dentine without the need of labeling.

  6. Multiphoton excitation spectra in biological samples

    Dickinson, Mary E.; Simbuerger, Eva; Zimmermann, Bernhard; Waters, Christopher W.; Fraser, Scott E.

    2003-01-01

    Multiphoton microscopy is becoming a popular mode of live and fixed cell imaging. This mode of imaging offers several advantages due to the fact that fluorochrome excitation is a nonlinear event resulting in excitation only at the plane of focus. Multiphoton excitation is enhanced by the use of ultrafast lasers emitting in the near IR, offering better depth penetration coupled with efficient excitation. Because these lasers, such as titanium:sapphire lasers, offer tunable output it is possibl...

  7. Multiphoton processes: conference proceedings

    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

  8. In-vitro visualization of corneal wound healing in an organ culture model using multiphoton autofluorescence and second harmonic generation microscopy

    Lo, Wen; Chang, Yuh-Ling; Sun, Yen; Lin, Sung-Jan; Jee, Shiou-Hwa; Tan, Hsin-Yuan; Dong, Chen-Yuan

    2007-02-01

    The aim of this work is to image the wound healing process of cornea in an in vitro organ culture model with noninvasive multiphoton imaging modality. Autofluorescence and second harmonic generation (SHG) were respectively used to monitor the alterations of cellular and collagenous components during wound healing processes. Within additional developments, this approach may be applied to in vivo visualization of corneal structural destruction and the subsequent regeneration.

  9. Multiphoton processes: conference proceedings

    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)

  10. Multiphoton bibliography, 1980

    This bibliography expands the collection of references appearing in multiphoton bibliography 1970-76, edited by J.H. Eberly and B. Karczewski, and the multiphoton bibliographies 1977, 1978, and 1979, edited by J.H. Eberly, J.W. Gallagher and E.C. Beaty. Items are included based on a list of words and phrases (e.g., multiphoton, intense field, strong field, multiple photon, 2-photon, 3-photon, etc.) or their equivalents appearing in an article's title or abstract

  11. Multiphoton ionization of atoms

    The paper is devoted to the analysis of high intensity effects which result from multiphoton ionization of atoms in a high laser intensity, ranging from 1010 to 1015 W cm-2. Resonant multiphoton ionization of atoms, the production of multiply charged ions, and electron energy spectra, are all discussed. (U.K.)

  12. Optical physics enables advances in multiphoton imaging

    Since the initial images were taken using a multiphoton imaging technique the method has rapidly established itself as the preferred method for imaging deeply into biological samples with micron resolution in three dimensions. Multiphoton imaging has thus enabled researchers in the life sciences to undertake studies that had previously been believed to be impossible without significantly perturbing the sample. Many of these experiments have only been realized due to close cooperation between optical physicists, from a range of disciplines, and the biomedical researchers. This paper will provide a general review of the current state of the field demonstrating how the various aspects of the physics development have brought the multiphoton technique to its current position at the forefront of biological microscopy. (topical review)

  13. A pragmatic guide to multiphoton microscope design

    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.

  14. MULTIPHOTON IONIZATION OF ATOMS

    Mainfray, G.

    1985-01-01

    Multiphoton ionization of one-electron atoms, such as atomic hydrogen and alkaline atoms, is well understood and correctly described by rigorous theoretical models. The present paper will be devoted to collisionless multiphoton ionization of many-electron atoms as rare gases. It induces removal of several electrons and the production of multiply charged ions. Up to Xe5+ ions are produced in Xe atoms. Doubly charged ions can be produced, either by simultaneous excitation of two electrons, or b...

  15. Multiphoton Squeezed States

    YANGXiao-Xue; WUYing

    2003-01-01

    We present analytical results for the multiphoton squeezed states defined through nonlinear quadrature-dependent Bogoliubov transformations. These analytical results turn a nonlinear problem into an essentially linear one and they can be utilized to express explicitly the mean walues and deviations of the quadrature operators and the photon variables under the multiphoton states in terms of those quantities averaged over the standard squeezed states which only involves the quadrature-independent Bogoliubov transformation.

  16. Multiphoton Squeezed States

    YANG Xiao-Xue; WU Ying

    2003-01-01

    We present analytical results for the multiphoton squeezed states defined through nonlinear quadraturedependent Bogoliubov transformations. These analytical results turn a nonlinear problem into an essentially linear one and they can be utilized to express explicitly the mean values and deviations of the quadrature operators and the photon variables under the multiphoton states in terms of those quantities averaged over the standard squeezed states which only involves the quadrature-independent Bogoliubov transformation.

  17. Moxifloxacin: Clinically compatible contrast agent for multiphoton imaging

    Taejun Wang; Won Hyuk Jang; Seunghun Lee; Yoon, Calvin J.; Jun Ho Lee; Bumju Kim; Sekyu Hwang; Chun-Pyo Hong; Yeoreum Yoon; Gilgu Lee; Viet-Hoan Le; Seoyeon Bok; G-One Ahn; Jaewook Lee; Yong Song Gho

    2016-01-01

    Multiphoton microscopy (MPM) is a nonlinear fluorescence microscopic technique widely used for cellular imaging of thick tissues and live animals in biological studies. However, MPM application to human tissues is limited by weak endogenous fluorescence in tissue and cytotoxicity of exogenous probes. Herein, we describe the applications of moxifloxacin, an FDA-approved antibiotic, as a cell-labeling agent for MPM. Moxifloxacin has bright intrinsic multiphoton fluorescence, good tissue penetra...

  18. DMD-based random-access optical-resolution photoacoustic microscopy

    Liang, Jinyang; Zhou, Yong; Winkler, Amy W.; Wang, Lidai; Maslov, Konstantin I.; Li, Chiye; Wang, Lihong V.

    2014-03-01

    The scanning mechanism is a major technical focus in optical-resolution photoacoustic microscopy. Flexible scanning access with fast scanning speed is desired to monitor biological and physiological dynamics with high temporal resolution. We developed random-access optical-resolution photoacoustic microscopy (RA-OR-PAM) using a digital micromirror device (DMD). Each micromirror on the DMD can be independently controlled, allowing imaging of regions of interest with arbitrary user-selected shapes without extraneous information. A global structural image is first acquired, and the regions of interest are selected. The laser beam then scans these regions exclusively, resulting in a faster frame rate than in a conventional raster scan. This system can rapidly scan arbitrarily shaped regions of interest with a lateral resolution of 3.6 μm within a 40×40 μm2 imaging area, a size comparable to the focal spot size of a 50 MHz ultrasound transducer. We demonstrated the random-access ability of RA-OR-PAM by imaging a monolayer of red blood cells. This system was then used to monitor blood flow in vivo within user-selected capillaries in a mouse ear. By imaging only the capillary of interest, the frame rate was increased by up to 13.3 times.

  19. Label-free assessment of adipose-derived stem cell differentiation using coherent anti-Stokes Raman scattering and multiphoton microscopy

    Mouras, Rabah; Bagnaninchi, Pierre O.; Downes, Andrew R; Elfick, Alistair P D

    2012-01-01

    ABSTRACT. Adult stem cells (SCs) hold great potential as likely candidates for disease therapy but also as sources of differentiated human cells in vitro models of disease. In both cases, the label-free assessment of SC differentiation state is highly desirable, either as a quality-control technology ensuring cells to be used clinically are of the desired lineage or to facilitate in vitro time-course studies of cell differentiation. We investigate the potential of nonlinear optical microscopy...

  20. Label-free assessment of adipose-derived stem cell differentiation using coherent anti-Stokes Raman scattering and multiphoton microscopy

    Mouras, Rabah; Bagnaninchi, Pierre O.; Downes, Andrew R.; Elfick, Alistair P. D.

    2012-11-01

    Adult stem cells (SCs) hold great potential as likely candidates for disease therapy but also as sources of differentiated human cells in vitro models of disease. In both cases, the label-free assessment of SC differentiation state is highly desirable, either as a quality-control technology ensuring cells to be used clinically are of the desired lineage or to facilitate in vitro time-course studies of cell differentiation. We investigate the potential of nonlinear optical microscopy as a minimally invasive technology to monitor the differentiation of adipose-derived stem cells (ADSCs) into adipocytes and osteoblasts. The induction of ADSCs toward these two different cell lineages was monitored simultaneously using coherent anti-Stokes Raman scattering, two photon excitation fluorescence (TPEF), and second harmonic generation at different time points. Changes in the cell's morphology, together with the appearance of biochemical markers of cell maturity were observed, such as lipid droplet accumulation for adipo-induced cells and the formation of extra-cellular matrix for osteo-induced cells. In addition, TPEF of flavoproteins was identified as a proxy for changes in cell metabolism that occurred throughout ADSC differentiation toward both osteoblasts and adipocytes. These results indicate that multimodal microscopy has significant potential as an enabling technology for the label-free investigation of SC differentiation.

  1. Multiphoton ionization of atoms

    This article provides an overview of the current understanding of multiphoton ionization of atoms. It begins with an introductory section to explain the background of the subject. Then the article develops the three topics which have been central themes of discussion in multiphoton ionization of atoms these past few years: multiply charged ion production, very high order harmonic generation, and above-threshold ionization, a name given to the absorption of a very large number of photons by an already ionized electron. A large part of the review is devoted to some theoretical aspects of multiphoton ionization of atoms and especially non-perturbative theories. Finally the article considers the very near future prospects of laser-electron interactions and more generally laser-matter interactions at 1018 -1019 W cm-2, an intensity range now within reach due to new short pulse laser technology. (author)

  2. Preparation of resistance random access memory samples for in situ transmission electron microscopy experiments

    Kudo, Masaki, E-mail: masakikudo@ist.hokudai.ac.jp; Arita, Masashi; Ohno, Yuuki; Fujii, Takashi; Hamada, Kouichi; Takahashi, Yasuo

    2013-04-30

    The ion-shadow method, an ion milling process using carbon particles as the mask material, is investigated as a means of preparing resistance random access memory (ReRAM) samples for in situ transmission electron microscopy (TEM). With a milling time of 1 hour (Ar{sup +}, 5 kV, 1 mA), multiple long needles (> 5 μm), on which there are miniaturized ReRAM devices comprising a ReRAM insulating layer sandwiched by two metallic electrodes, are formed on the substrate. Device sizes of up to several hundreds of nm are easily obtained with the method. The internal part of small devices (i.e., up to 100 nm) can be observed by TEM. Electrical measurements using an in situ TEM holder demonstrate that sufficient electric contact is obtained without any electric shortage between the electrodes due to re-deposition of milled material. The ion-shadow method is confirmed to be a quick and easy method suitable for in situ TEM experiments, especially for ReRAM devices which are highly susceptible to destruction during the switching operation. - Highlights: ► Preparation method of in situ transmission electron microscopy samples is shown. ► Long and sharp needle-shaped samples are formed with the ion-shadow method. ► Resistive switching samples are formed at the tips of needle-shaped samples. ► Current–voltage characteristics obtained in an electron microscope are shown.

  3. Preparation of resistance random access memory samples for in situ transmission electron microscopy experiments

    The ion-shadow method, an ion milling process using carbon particles as the mask material, is investigated as a means of preparing resistance random access memory (ReRAM) samples for in situ transmission electron microscopy (TEM). With a milling time of 1 hour (Ar+, 5 kV, 1 mA), multiple long needles (> 5 μm), on which there are miniaturized ReRAM devices comprising a ReRAM insulating layer sandwiched by two metallic electrodes, are formed on the substrate. Device sizes of up to several hundreds of nm are easily obtained with the method. The internal part of small devices (i.e., up to 100 nm) can be observed by TEM. Electrical measurements using an in situ TEM holder demonstrate that sufficient electric contact is obtained without any electric shortage between the electrodes due to re-deposition of milled material. The ion-shadow method is confirmed to be a quick and easy method suitable for in situ TEM experiments, especially for ReRAM devices which are highly susceptible to destruction during the switching operation. - Highlights: ► Preparation method of in situ transmission electron microscopy samples is shown. ► Long and sharp needle-shaped samples are formed with the ion-shadow method. ► Resistive switching samples are formed at the tips of needle-shaped samples. ► Current–voltage characteristics obtained in an electron microscope are shown

  4. Clinical multiphoton FLIM tomography

    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.

  5. Readily Accessible Multiplane Microscopy: 3D Tracking the HIV-1 Genome in Living Cells.

    Itano, Michelle S; Bleck, Marina; Johnson, Daniel S; Simon, Sanford M

    2016-02-01

    Human immunodeficiency virus (HIV)-1 infection and the associated disease AIDS are a major cause of human death worldwide with no vaccine or cure available. The trafficking of HIV-1 RNAs from sites of synthesis in the nucleus, through the cytoplasm, to sites of assembly at the plasma membrane are critical steps in HIV-1 viral replication, but are not well characterized. Here we present a broadly accessible microscopy method that captures multiple focal planes simultaneously, which allows us to image the trafficking of HIV-1 genomic RNAs with high precision. This method utilizes a customization of a commercial multichannel emission splitter that enables high-resolution 3D imaging with single-macromolecule sensitivity. We show with high temporal and spatial resolution that HIV-1 genomic RNAs are most mobile in the cytosol, and undergo confined mobility at sites along the nuclear envelope and in the nucleus and nucleolus. These provide important insights regarding the mechanism by which the HIV-1 RNA genome is transported to the sites of assembly of nascent virions. PMID:26567131

  6. Multiphoton ionization of polarized atoms

    A theory is derived for the multiphoton ionization of polarized atoms. The angular distributions of the differential and total ionization probabilities are studied for various polarizations of the electromagnetic radiation. The circular dichroism is also studied. The multiphoton ionization of oriented s-state atoms near a resonance is studied separately. Some relevant experiments which might be carried out are discussed

  7. Multiphoton ionization of polarized atoms

    A theory of multiphoton ionization of polarized atoms is developed. The angular dependences of the different and total ionization probabilities for various polarizations of the radiation and the circular dichroism effect are investigated. Multiphoton ionization of oriented in the s-states near resonance is considered separately

  8. Optical clearing and multiphoton imaging of paraffin-embedded specimens

    Wilson, Jesse W.; Degan, Simone; Fischer, Martin C.; Warren, Warren S.

    2013-02-01

    New labeling, imaging, or analysis tools could provide new retrospective insights when applied to archived, paraffin-embedded samples. Deep-tissue multiphoton microscopy of paraffin-embedded specimens is achieved using optical clearing with mineral oil. We tested a variety of murine tissue specimens including skin, lung, spleen, kidney, and heart, acquiring multiphoton autofluorescence and second-harmonic generation, and pump-probe images This technique introduces the capability for non-destructive 3-dimensional microscopic imaging of existing archived pathology specimens, enabling retrospective studies.

  9. Multiphoton imaging with high peak power VECSELs

    Mirkhanov, Shamil; Quarterman, Adrian H.; Swift, Samuel; Praveen, Bavishna B.; Smyth, Conor J. C.; Wilcox, Keith G.

    2016-03-01

    Multiphoton imaging (MMPI) has become one of thee key non-invasive light microscopy techniques. This technique allows deep tissue imaging with high resolution and less photo-damage than conventional confocal microscopy. MPI is type of laser-scanning microscopy that employs localized nonlinear excitation, so that fluorescence is excited only with is scanned focal volume. For many years, Ti: sapphire femtosecond lasers have been the leading light sources for MPI applications. However, recent developments in laser sources and new types of fluorophores indicate that longer wavelength excitation could be a good alternative for these applications. Mode-locked VECSEELs have the potential to be low cost, compact light sources for MPI systems, with the additional advantage of broad wavelength coverage through use of different semiconductor material systems. Here, we use a femtosecond fibber laser to investigate the effect average power and repetition rate has on MPI image quality, to allow us to optimize our mode-locked VVECSELs for MPI.

  10. A novel flexible clinical multiphoton tomograph for early melanoma detection, skin analysis, testing of anti-age products, and in situ nanoparticle tracking

    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.

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

    König, Karsten; Andersen, Peter E.; Le, Tuan; Breunig, Hans Georg

    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...

  12. Calculation of multiphoton ionization processes

    Chang, T. N.; Poe, R. T.

    1976-01-01

    We propose an accurate and efficient procedure in the calculation of multiphoton ionization processes. In addition to the calculational advantage, this procedure also enables us to study the relative contributions of the resonant and nonresonant intermediate states.

  13. Multiphoton ionization of Uracil

    Prieto, Eladio; Martinez, Denhi; Guerrero, Alfonso; Alvarez, Ignacio; Cisneros, Carmen

    2016-05-01

    Multiphoton ionization and dissociation of Uracil using a Reflectron time of flight spectrometer was performed along with radiation from the second harmonic of a Nd:YAG laser. Uracil is one of the four nitrogen bases that belong to RNA. The last years special interest has been concentrated on the study of the effects under UV radiation in nucleic acids1 and also in the role that this molecule could have played in the origin and development of life on our planet.2 The MPI mass spectra show that the presence and intensity of the resulting ions strongly depend on the density power. The identification of the ions in the mass spectra is presented. The results are compared with those obtained in other laboratories under different experimental conditions and some of them show partial agreement.3 The present work was supported by CONACYT-Mexico Grant 165410 and DGAPA UNAM Grant IN101215 and IN102613.

  14. Enabling Multiphoton and Second Harmonic Generation Imaging in Paraffin-Embedded and Histologically Stained Sections

    Monaghan, Michael G.; Kroll, Sebastian; Brucker, Sara Y.; Schenke-Layland, Katja

    2016-01-01

    Nonlinear microscopy, namely multiphoton imaging and second harmonic generation (SHG), is an established noninvasive technique useful for the imaging of extracellular matrix (ECM). Typically, measurements are performed in vivo on freshly excised tissues or biopsies. In this article, we describe the effect of rehydrating paraffin-embedded sections on multiphoton and SHG emission signals and the acquisition of nonlinear images from hematoxylin and eosin (H&E)-stained sections before and after a...

  15. Multiphoton imaging with a nanosecond supercontinuum source

    Lefort, Claire; O'Connor, Rodney P.; Blanquet, Véronique; Baraige, Fabienne; Tombelaine, Vincent; Lévêque, Philippe; Couderc, Vincent; Leproux, Philippe

    2016-03-01

    Multiphoton microscopy is a well-established technique for biological imaging of several kinds of targets. It is classically based on multiphoton processes allowing two means of contrast simultaneously: two-photon fluorescence (TPF) and second harmonic generation (SHG). Today, the quasi exclusive laser technology used in that aim is femtosecond titanium sapphire (Ti: Sa) laser. We experimentally demonstrate that a nanosecond supercontinuum laser source (STM-250-VIS-IR-custom, Leukos, France; 1 ns, 600-2400 nm, 250 kHz, 1 W) allows to obtain the same kind of image quality in the case of both TPF and SHG, since it is properly filtered. The first set of images concerns the muscle of a mouse. It highlights the simultaneous detection of TPF and SHG. TPF is obtained thanks to the labelling of alpha-actinin with Alexa Fluor® 546 by immunochemistry. SHG is created from the non-centrosymmetric organization of myosin. As expected, discs of actin and myosin are superimposed alternatively. The resulting images are compared with those obtained from a standard femtosecond Ti: Sa source. The physical parameters of the supercontinuum are discussed. Finally, all the interest of using an ultra-broadband source is presented with images obtained in vivo on the brain of a mouse where tumor cells labeled with eGFP are grafted. Texas Red® conjugating Dextran is injected into the blood vessels network. Thus, two fluorophores having absorption wavelengths separated by 80 nm are imaged simultaneously with a single laser source.

  16. Super-resolving multi-photon interferences with independent light sources

    Oppel, Steffen; Kok, Pieter; von Zanthier, Joachim

    2012-01-01

    Multi-photon interferences with indistinguishable photons from independent light sources are at the focus of current research owing to their potential in optical quantum computing, creating remote entanglement for quantum computation and communication, and quantum metrology. The paradigmatic states for multi-photon interference are the highly entangled NOON states, which can be used to achieve increased resolution in spectroscopy, interferometry, lithography, and microscopy. Multi-photon interferences from independent, uncorrelated emitters can also lead to enhanced resolution in metrology and imaging. So far, such interferences have been observed with maximally two independent emitters. Here, we report multi-photon interferences with up to five independent emitters, displaying interference patterns equivalent to those of NOON states. Experimental results with independent thermal light sources confirm this NOON-like modulation. The experiment is an extension of the landmark measurement by Hanbury Brown and Tw...

  17. Multiphotons and Photon-Jets

    Toro, Natalia; Yavin, Itay(Perimeter Institute for Theoretical Physics, Waterloo, ON, Canada)

    2012-01-01

    We discuss an extension of the Standard Model with a new vector-boson decaying predominantly into a multi-photon final state through intermediate light degrees of freedom. The model has a distinctive phase in which the photons are collimated. As such, they would fail the isolation requirements of standard multi-photon searches, but group naturally into a novel object, the photon-jet. Once defined, the photon-jet object facilitates more inclusive searches for similar phenomena. We present a co...

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

    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 1016 W/cm2. In all, the research has resulted in 36 publications (five journal covers) and nine invention disclosures, five of which have continued on to patenting

  19. Analysis of the metabolic deterioration of ex vivo skin from ischemic necrosis through the imaging of intracellular NAD(P)H by multiphoton tomography and fluorescence lifetime imaging microscopy

    Sanchez, Washington Y.; Prow, Tarl W.; Sanchez, Washington H.; Grice, Jeffrey E.; Roberts, Michael S.

    2010-07-01

    Ex vivo human skin has been used extensively for cosmeceutical and drug delivery studies, transplantable skin allografts, or skin flaps. However, it has a half-life of a few days due to ischemic necrosis. Traditional methods of assessing viability can be time-consuming and provide limited metabolic information. Using multiphoton tomography and fluorescence lifetime imaging (MPT-FLIM) we assess ischemic necrosis of ex vivo skin by NAD(P)H autofluorescence intensity and fluorescence lifetime. Ex vivo skin is stored in the presence and absence of nutrient media (Dulbecco Modified Eagle Medium) at -20, 4, and 37 °C and room temperature over a 7-day time course to establish different rates of metabolic deterioration. At higher temperatures we observe a decrease in NAD(P)H autofluorescence, higher image noise, and a significant increase in the average fluorescence lifetime (τm) from ~1000 to 2000 ps. Additionally, significant distortions in NAD(P)H fluorescence lifetime histograms correspond to the reduction in autofluorescence. Skin kept at 4 °C, with or without media, showed the least change. Our findings suggest that MPT-FLIM enables useful noninvasive optical biopsies to monitor the metabolic state and deterioration of human skin for research and clinical purposes.

  20. Moxifloxacin: Clinically compatible contrast agent for multiphoton imaging

    Wang, Taejun; Jang, Won Hyuk; Lee, Seunghun; Yoon, Calvin J.; Lee, Jun Ho; Kim, Bumju; Hwang, Sekyu; Hong, Chun-Pyo; Yoon, Yeoreum; Lee, Gilgu; Le, Viet-Hoan; Bok, Seoyeon; Ahn, G.-One; Lee, Jaewook; Gho, Yong Song; Chung, Euiheon; Kim, Sungjee; Jang, Myoung Ho; Myung, Seung-Jae; Kim, Myoung Joon; So, Peter T. C.; Kim, Ki Hean

    2016-06-01

    Multiphoton microscopy (MPM) is a nonlinear fluorescence microscopic technique widely used for cellular imaging of thick tissues and live animals in biological studies. However, MPM application to human tissues is limited by weak endogenous fluorescence in tissue and cytotoxicity of exogenous probes. Herein, we describe the applications of moxifloxacin, an FDA-approved antibiotic, as a cell-labeling agent for MPM. Moxifloxacin has bright intrinsic multiphoton fluorescence, good tissue penetration and high intracellular concentration. MPM with moxifloxacin was demonstrated in various cell lines, and animal tissues of cornea, skin, small intestine and bladder. Clinical application is promising since imaging based on moxifloxacin labeling could be 10 times faster than imaging based on endogenous fluorescence.

  1. High-order multiphoton ionization photoelectron spectroscopy of NO

    Photoelectron energy and 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 A2Σ+ (v=0) level yielded a distribution of electron energies corresponding to all accessible vibrational levels (v+=0--6) of the nascent ion. Angular distributions of electrons corresponding to v+=0 and v+=3 were significantly different. The 3+2 REMPI via the A2Σ+ (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

  2. Multiphoton quantum optics and quantum state engineering

    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

    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.

  4. Invited Review Article: Pump-probe microscopy

    Fischer, Martin C.; Wilson, Jesse W.; Robles, Francisco E.; Warren, Warren S.

    2016-03-01

    Multiphoton microscopy has rapidly gained popularity in biomedical imaging and materials science because of its ability to provide three-dimensional images at high spatial and temporal resolution even in optically scattering environments. Currently the majority of commercial and home-built devices are based on two-photon fluorescence and harmonic generation contrast. These two contrast mechanisms are relatively easy to measure but can access only a limited range of endogenous targets. Recent developments in fast laser pulse generation, pulse shaping, and detection technology have made accessible a wide range of optical contrasts that utilize multiple pulses of different colors. Molecular excitation with multiple pulses offers a large number of adjustable parameters. For example, in two-pulse pump-probe microscopy, one can vary the wavelength of each excitation pulse, the detection wavelength, the timing between the excitation pulses, and the detection gating window after excitation. Such a large parameter space can provide much greater molecular specificity than existing single-color techniques and allow for structural and functional imaging without the need for exogenous dyes and labels, which might interfere with the system under study. In this review, we provide a tutorial overview, covering principles of pump-probe microscopy and experimental setup, challenges associated with signal detection and data processing, and an overview of applications.

  5. Invited Review Article: Pump-probe microscopy.

    Fischer, Martin C; Wilson, Jesse W; Robles, Francisco E; Warren, Warren S

    2016-03-01

    Multiphoton microscopy has rapidly gained popularity in biomedical imaging and materials science because of its ability to provide three-dimensional images at high spatial and temporal resolution even in optically scattering environments. Currently the majority of commercial and home-built devices are based on two-photon fluorescence and harmonic generation contrast. These two contrast mechanisms are relatively easy to measure but can access only a limited range of endogenous targets. Recent developments in fast laser pulse generation, pulse shaping, and detection technology have made accessible a wide range of optical contrasts that utilize multiple pulses of different colors. Molecular excitation with multiple pulses offers a large number of adjustable parameters. For example, in two-pulse pump-probe microscopy, one can vary the wavelength of each excitation pulse, the detection wavelength, the timing between the excitation pulses, and the detection gating window after excitation. Such a large parameter space can provide much greater molecular specificity than existing single-color techniques and allow for structural and functional imaging without the need for exogenous dyes and labels, which might interfere with the system under study. In this review, we provide a tutorial overview, covering principles of pump-probe microscopy and experimental setup, challenges associated with signal detection and data processing, and an overview of applications. PMID:27036751

  6. Multiphoton spectroscopy in heavy elements

    Solarz, R.W.; Paisner, J.A.; Worden, E.F.

    1977-05-03

    Some recently discovered regularities in the spectra of heavy elements which are also applicable to the analysis of the spectra of lighter atoms are described. It is pointed out that stepwise resonant multiphoton methods are irreplaceable tools in the study of high lying states in a complex atomic system. Systematic applications of these methods has permitted regularities to be observed which also hold for the lighter elements. It is noted that greatly increased understanding of the excited state structure of heavy atoms is not possible. 8 references. (JFP)

  7. Multiphoton dissociation of polyatomic molecules

    The dynamics of infrared multiphoton excitation and dissociation of SF6 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 SF6, the pulse duration of the CO2 laser and the frequency in both one and two laser experiments. Translational energy distributions of the SF5 dissociation product measured by time of flight and angular distributions and the dissociation lifetime of excited SF6 as inferred from the observation of secondary dissociation of SF5 into SF4 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

  8. Multiphoton Quantum Optics and Quantum State Engineering

    Dell'Anno, F; Illuminati, F; 10.1016/j.physrep.2006.01.004

    2009-01-01

    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. 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...

  9. MULTI-PHOTON PHOSPHOR FEASIBILITY RESEARCH

    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

  10. ATOMS INTERACTING WITH ELECTROMAGNETIC FIELDS, MULTIPHOTON IONIZATION

    Mainfray, G

    1982-01-01

    The non linear interaction between an intense laser radiation and atoms leads to ionization through the absorption of N photons from the laser radiation via laser-induced virtual states. The multiphoton ionization rate varies as a function of the laser intensity I as IN. We discuss the two most important effects which govern multiphoton ionization processes : resonance effects and laser-coherence effects. In a moderate laser intensity range (107 - 109 W cm-2) corresponding to the two, three o...

  11. Quantum electrodynamic perspective on multiphoton ionization

    A fully quantum nonperturbative method is developed to describe multiphoton ionization in intense fields. It is shown that, treating the radiation field with quantum electrodynamic (QED) theory enables the authors to obtain the above-threshold ionization energy distribution spectrum in analytical form firstly. Moreover, in addition to the well-known semiclassical theory, the framework presented here, derived from a QED perspective, provides a new picture of the multiphoton ionization

  12. Multiphoton Quantum Optics and Quantum State Engineering

    Dell'Anno, F.; Siena, S; Illuminati, F.

    2007-01-01

    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 ...

  13. Multiphoton ionization of uranium hexafluoride

    Multiphoton ionization (MPI) time-of-flight mass spectroscopy (TOFMS) and photoelectron spectroscopy (PES) studies of UF6 are reported 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. In general, the doubly charged uranium ion (U2+) intensity is much greater than that of the singly charged uranium ion (U+). For the case of the tunable dye laser experiments, the Un+ (n=1--4) wavelength dependence is relatively unstructured and does not show observable resonance enhancement at known atomic uranium excitation wavelengths. The MPI-PES studies reveal only very slow electrons (≤0.5 eV) for all wavelengths investigated. The dominance of the U2+ ion, the absence or very small intensities of UF+x (x=1--3) fragments, the unstructured wavelength dependence, and the preponderance of slow electrons all indicate that mechanisms may exist other than ionization of bare U atoms following the stepwise photodissociation of F atoms from the parent molecule. The data also argue against stepwise photodissociation of UF+x (x=5,6) ions. Neither of the traditional MPI mechanisms (''neutral ladder'' or the ''ionic ladder'') are believed to adequately describe the ionization phenomena observed. We propose that the multiphoton excitation of UF6 under these experimental conditions results in a highly excited molecule, superexcited UF6**

  14. Multiphoton ionization and multiphoton resonances in the tunneling regime

    The rate of ionization of an atom of helium, argon, or hydrogen exposed to an intense monochromatic laser field and the quasienergy spectrum of their dressed states are studied for values of the Keldysh parameter between 1 and 0.6 and wavelengths between 390 and 1300 nm. The calculations are carried out within the non-Hermitian Floquet theory. Resonances with intermediate excited states significantly affect ionization from the dressed ground state at all the intensities and all the wavelengths considered. The dressed excited states responsible for these structures are large-α0 states akin to the Kramers-Henneberger states of the high-frequency Floquet theory. Within the single-active-electron approximation, these large-α0 states become species independent at sufficiently high intensity or sufficiently long wavelength. Apart for the resonance structures arising from multiphoton coupling with excited states, the ab initio Floquet ionization rate is in excellent agreement with the predictions of two different calculations in the strong field approximation, one based on a length-gauge formulation of this approximation and one based on a velocity-gauge formulation. The calculations also confirm the validity of the ω2 expansion as an alternative to the strong field approximation for taking into account the nonadiabaticity of the ionization process in intense low-frequency laser fields.

  15. Multiphoton ionization of uranium hexafluoride

    Armstrong, D. P.; Harkins, D. A.; Compton, R. N.; Ding, D.

    1994-01-01

    Multiphoton ionization (MPI) time-of-flight mass spectroscopy (TOFMS) and photoelectron spectroscopy (PES) studies of UF6 are reported 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. In general, the doubly charged uranium ion (U2+) intensity is much greater than that of the singly charged uranium ion (U+). For the case of the tunable dye laser experiments, the Un+ (n=1-4) wavelength dependence is relatively unstructured and does not show observable resonance enhancement at known atomic uranium excitation wavelengths. The MPI-PES studies reveal only very slow electrons (≤0.5 eV) for all wavelengths investigated. The dominance of the U2+ ion, the absence or very small intensities of UF+x (x=1-3) fragments, the unstructured wavelength dependence, and the preponderance of slow electrons all indicate that mechanisms may exist other than ionization of bare U atoms following the stepwise photodissociation of F atoms from the parent molecule. The data also argue against stepwise photodissociation of UF+x (x=5,6) ions. Neither of the traditional MPI mechanisms (``neutral ladder'' or the ``ionic ladder'') are believed to adequately describe the ionization phenomena observed. We propose that the multiphoton excitation of UF6 under these experimental conditions results in a highly excited molecule, superexcited UF6**. The excitation of highly excited UF6** is proposed to be facilitated by the well known ``giant resonance,'' whose energy level lies in the range of 12-14 eV above that of ground state UF6. The highly excited molecule then primarily dissociates, via multiple channels, into Un+, UF+x, fluorine atoms, and ``slow'' electrons, although dissociation

  16. Stepwise multi-photon activation fluorescence reveals a new method of melanoma imaging for dermatologists

    Lai, Zhenhua; Lian, Christine; Ma, Jie; Yu, Jingyi; Gu, Zetong; Rajadhyaksha, Milind; DiMarzio, Charles A.

    2014-02-01

    Previous research has shown that the stepwise multi-photon activated fluorescence (SMPAF) of melanin, activated by a continuous-wave (CW) mode near infrared (NIR) laser, is a low cost and reliable method of detecting melanin. SMPAF images of melanin in a mouse hair and a formalin fixed mouse melanoma were compared with conventional multiphoton fluorescence microscopy (MPFM) images and confocal reflectance microscopy (CRM) images, all of which were acquired at an excitation wavelength of 920 nm, to further prove the effectiveness of SMPAF in detecting melanin. SMPAF images add specificity for melanin detection to MPFM images and CRM images. Melanin SMPAF can be a promising technology to enable melanoma imaging for dermatologists.

  17. Multiphoton ionization of H- and He in intense laser fields

    The recently proposed R-matrix-Floquet theory of multiphoton processes has been used to calculate multiphoton ionization rates for the two-electron systems H- and He in intense laser fields. The theory is nonperturbative and includes electron-electron correlations. Results are presented for total and partial multiphoton ionization rates and novel nonperturbative correlation effects are discussed

  18. Multiphoton ionization of atomic cesium

    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

  19. Models for multiphoton ionization processes

    Using the Fourier transform method, several analytical models for multiphoton processes have been developed, which emphasize the non-perturbative regime of C-C transitions. It was found that the population trapping of continuum-continuum (C-C) transitions can occurs under general conditions: for two continua with energy-dependent matrix elements, and for an infinite number of structureless continua with coupled bands. A above-threshold ionization model with two-frequency smooth pulses for hydrogenic atom was first proposed. This model give a simple form for analytical solutions if an ionization threshold is negligible. The results predict that (a) there is no trapping or saturation of the bound-continuum transition and that the ionization rate is independent of the redistribution among the continua for the C-C matrix element chosen; (b) There are peak switch in photoelectron energy spectra which depends on not only redistribution laser intensity, but also the pulses shape and their overlap in time, as well as the coupling between the bound state and continua; (c) peaks about the ionization energy in the photoelectron spectra are symmetric and only appear during the ionization process; (d) as the laser intensity is increased, the total angular photoelectrons distribution peak strongly in forward and backward direction of the redistribution laser polarization; and the minima of this distribution are not zero for higher laser intensity. The photon spectrum is also investigated

  20. Infrared multiphoton absorption and decomposition

    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

  1. Multiphoton imaging to identify grana, stroma thylakoid, and starch inside an intact leaf

    Chen, Mei-Yu; Zhuo, Guan-Yu; Chen, Kuan-Chieh; Wu, Pei-Chun; Hsieh, Tsung-Yuan; Liu, Tzu-Ming; Chu, Shi-Wei

    2014-01-01

    Background Grana and starch are major functional structures for photosynthesis and energy storage of plant, respectively. Both exhibit highly ordered molecular structures and appear as micrometer-sized granules inside chloroplasts. In order to distinguish grana and starch, we used multiphoton microscopy, with simultaneous acquisition of two-photon fluorescence (2PF) and second harmonic generation (SHG) signals. SHG is sensitive to crystallized structures while 2PF selectively reveals the dist...

  2. Multiphoton polymerization using optical trap assisted nanopatterning

    Leitz, Karl-Heinz; Tsai, Yu-Cheng; Flad, Florian; Schäffer, Eike; Quentin, Ulf; Alexeev, Ilya; Fardel, Romain; Arnold, Craig B.; Schmidt, Michael

    2013-06-01

    In this letter, we show the combination of multiphoton polymerization and optical trap assisted nanopatterning (OTAN) for the additive manufacturing of structures with nanometer resolution. User-defined patterns of polymer nanostructures are deposited on a glass substrate by a 3.5 μm polystyrene sphere focusing IR femtosecond laser pulses, showing minimum feature sizes of λ/10. Feature size depends on the applied laser fluence and the bead surface spacing. A finite element model describes the intensity enhancement in the microbead focus. The results presented suggest that OTAN in combination with multiphoton processing is a viable technique for additive nanomanufacturing with sub-diffraction-limited resolution.

  3. Observation of amyloid precursor protein cleavage and Aβ generation in living cells by using multiphoton laser scanning microscopy%多光子激光扫描成像技术对活细胞内淀粉样前体蛋白裂解和β-淀粉样蛋白生成的观察

    李晓晴; 张苏明; 杨华静; 张智红

    2007-01-01

    Objective To investigate the proteolytic mechanism of amyloid precursor protein (APP) and to explore amyloidbeta (Aβ) generation in living neurons. Methods DNA fragments were amplified by PCR or synthesized. The four fragments- CFP- 54bp- YFP and C99 were ligated into pcDNAS.O vector to construct the recombinant plasmids pcDNA3.0-CFP-54bp-YFP and pcDNA3.0-CFP-54bp-YFP-C99. The SH-SY5Y cells were transiently transfected with pcDNA3.0-CFP-54bp-YFP or pcDNA3.0-CFP-54bp-YFP-C99.The SH-SY5Y cells were transiently transfected with pcDNA3.0-CFP-54bp-YFP or pcDNA3.0-CFP-54bp-YFP-C99.The expression of fusion gene was examined under a multiphoton laser scanning microscope.Fluorescence resonance energy transfer (FRET) was used to measure the p cleavage and y cleavage of APP.Aβ generation was confirmed by immunocytochemistry and multiphoton laser scanning microscopy.Cell viability was tested by MTT assay at different time points.Results (1) The double restriction endonuclease digestion and sequencing analysis confirmed the authenticity of the recombinant plasmids pcDNA3.0-CFP-54bp-YFP and pcDNA3.0-CFP-54bp-YFP-C99.(2) Blue and yellow fluorescences were detected in the transfected cells.(3) FRET occurred in pcDNA3.0-CFP-54bp-YFP-transfected cells but not in pcDNA3.0-CFP-54bp-YFP-C99-transfected cells.(4) Aβ was produced in the pcDNA3.0-CFP-54bp-YFP-C99 transfected cells.(5) Aβ-deposition was widespread in the cell.(6) Cell viability decreased along with the intracellular Aβ deposition.Conclusion C99 is important for the APP β cleavage.Aβ may be generated and deposited in cells at the early stage of Alzheimer's disease.Intracellular Aβ accumulation brings deleterious effects on cells.%目的 在活细胞内探究淀粉样前体蛋白(amyloid precursor protein,APP)的裂解和β-淀粉样蛋白(amyloid beta,Aβ)的生成机制.方法 利用PCR扩增CFP(编码蓝色荧光蛋白),YFP(编码黄色荧光蛋白)和C99(编码APP最后99个氨基酸)三片段.含有54

  4. Optical investigation of the intergrowth structure and accessibility of Brønsted acid sites in etched SSZ-13 zeolite crystals by confocal fluorescence microscopy.

    Sommer, Linn; Svelle, Stian; Lillerud, Karl Petter; Stöcker, Michael; Weckhuysen, Bert M; Olsbye, Unni

    2010-11-01

    Template decomposition followed by confocal fluorescence microscopy reveals a tetragonal-pyramidal intergrowth of subunits in micrometer-sized nearly cubic SSZ-13 zeolite crystals. In order to accentuate intergrowth boundaries and defect-rich areas within the individual large zeolite crystals, a treatment with an etching NaOH solution is applied. The defective areas are visualized by monitoring the spatial distribution of fluorescent tracer molecules within the individual SSZ-13 crystals by confocal fluorescence microscopy. These fluorescent tracer molecules are formed at the inner and outer crystal surfaces by utilizing the catalytic activity of the zeolite in the oligomerization reaction of styrene derivatives. This approach reveals various types of etching patterns that are an indication for the defectiveness of the studied crystals. We can show that specially one type of crystals, denoted as core-shell type, is highly accessible to the styrene molecules after etching. Despite the large crystal dimensions, the whole core-shell type SSZ-13 crystal is utilized for catalytic reaction. Furthermore, the confocal fluorescence microscopy measurements indicate a nonuniform distribution of the catalytically important Brønsted acid sites underlining the importance of space-resolved measurements. PMID:20496927

  5. MULTIPLE IONIZATION OF ATOMS THROUGH MULTIPHOTON ABSORPTION

    L'Huillier, A

    1987-01-01

    We review the main aspects of multiple ionization of rare gases in strong laser fields (above 1012W.cm-2). We discuss the mechanisms responsible for the multi-electron ejection : inner-shell ionization or outer-shell ionization, one-step or multi-step. We show the differences between one-photon and multi-photon multiple ionization.

  6. Route to Direct Multiphoton Multiple Ionization

    Lambropoulos, P; Papamihail, K G

    2011-01-01

    We address the concept of direct multiphoton multiple ionization in atoms exposed to intense, short wavelength radiation and explore the conditions under which such processes dominate over the sequential. Their contribution is shown to be quite robust, even under intensity fluctuations and interaction volume integration, and reasonable agreement with experimental data is also found.

  7. Route to direct multiphoton multiple ionization

    We address the concept of direct multiphoton multiple ionization in atoms exposed to intense, short-wavelength radiation and explore the conditions under which such processes dominate over the sequential. Their contribution is shown to be quite robust, even under intensity fluctuations and interaction volume integration, and reasonable agreement with experimental data is also found.

  8. Brain plasticity and functionality explored by nonlinear optical microscopy

    Sacconi, L.; Allegra, L.; Buffelli, M.; Cesare, P.; D'Angelo, E.; Gandolfi, D.; Grasselli, G.; Lotti, J.; Mapelli, J.; Strata, P.; Pavone, F. S.

    2010-02-01

    In combination with fluorescent protein (XFP) expression techniques, two-photon microscopy has become an indispensable tool to image cortical plasticity in living mice. In parallel to its application in imaging, multi-photon absorption has also been used as a tool for the dissection of single neurites with submicrometric precision without causing any visible collateral damage to the surrounding neuronal structures. In this work, multi-photon nanosurgery is applied to dissect single climbing fibers expressing GFP in the cerebellar cortex. The morphological consequences are then characterized with time lapse 3-dimensional two-photon imaging over a period of minutes to days after the procedure. Preliminary investigations show that the laser induced fiber dissection recalls a regenerative process in the fiber itself over a period of days. These results show the possibility of this innovative technique to investigate regenerative processes in adult brain. In parallel with imaging and manipulation technique, non-linear microscopy offers the opportunity to optically record electrical activity in intact neuronal networks. In this work, we combined the advantages of second-harmonic generation (SHG) with a random access (RA) excitation scheme to realize a new microscope (RASH) capable of optically recording fast membrane potential events occurring in a wide-field of view. The RASH microscope, in combination with bulk loading of tissue with FM4-64 dye, was used to simultaneously record electrical activity from clusters of Purkinje cells in acute cerebellar slices. Complex spikes, both synchronous and asynchronous, were optically recorded simultaneously across a given population of neurons. Spontaneous electrical activity was also monitored simultaneously in pairs of neurons, where action potentials were recorded without averaging across trials. These results show the strength of this technique in describing the temporal dynamics of neuronal assemblies, opening promising

  9. Multi-Photon Interference and Temporal Distinguishability of Photons

    Ou, Z. Y.

    2007-01-01

    A number of recent interference experiments involving multiple photons are reviewed. These experiments include generalized photon bunching effects, generalized Hong-Ou-Mandel interference effects and multi-photon interferometry for demonstrations of multi-photon de Broglie wavelength. The multi-photon states used in these experiments are from two pairs of photons in parametric down-conversion. We find that the size of the interference effect in these experiments, characterized by the visibili...

  10. Multiphoton ionization with femtosecond laser pulses

    There are a number of reasons to use subpicosecond pulses in multiphoton ionization experiments. Pulses with shorter risetimes make it possible to study processes with higher rates before one runs into the problem of depletion of target atoms. Furthermore, the momentum of the electron does not change between the point of ionization and the detector if the pulse expires before the electron has time to sample the spatial inhomogeneity of the light intensity. This makes it possible to identify the intensity at which an electron was formed from the energy with which it reaches the detector. Yet another advantage is the fact that the primary ionization products are subject to the ionizing radiation for only a short amount of time, thus reducing the probability of sequential ionization as compared to direct processes. In this paper the experimental results on the multiphoton ionization of xenon with pulses of 100 fs, that were obtained during the preceding year at ENSTA, Palaiseau, will be presented

  11. Multiphoton ionization/dissociation of osmium tetroxide

    The mechanisms leading to laser multiphoton ionization and dissociation (MPI/MPD) of osmium tetroxide (OsO4) have been investigated from measurements of the kinetic energies of product ions (Os+, Os2+, OsO+, O2+, O+) and photoelectrons as a function of the laser wavelength. Neutral channels, intermediate to the dominant Os+ ionization channel, such as OsO4→OsO4-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+

  12. In vivo multiphoton tomography of skin cancer

    König, Karsten; Riemann, Iris; Ehlers, Alexander; Buckle, Rainer; Dimitrow, Enrico; Kaatz, Martin; Fluhr, Joachim; Elsner, Peter

    2006-02-01

    The multiphoton tomograph DermaInspect was used to perform first clinical studies on the early non-invasive detection of skin cancer based on non-invasive optical sectioning of skin by two-photon autofluorescence and second harmonic generation. In particular, deep-tissue pigmented lesions -nevi- have been imaged with intracellular resolution using near infrared (NIR) femtosecond laser radiation. So far, more than 250 patients have been investigated. Cancerous tissues showed significant morphological differences compared to normal skin layers. In the case of malignant melanoma, the occurrence of luminescent melanocytes has been detected. Multiphoton tomography will become a novel non-invasive method to obtain high-resolution 3D optical biopsies for early cancer detection, treatment control, and in situ drug screening.

  13. Multiphoton processes in isolated atoms and molecules

    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

  14. Medium-induced multi-photon radiation

    MA, Hao; Salgado, Carlos A.; Tywoniuk, Konrad

    2011-01-01

    We study the spectrum of multi-photon radiation off a fast quark in medium in the BDMPS/ASW approach. We reproduce the medium-induced one-photon radiation spectrum in dipole approximation, and go on to calculate the two-photon radiation in the Moli\\`{e}re limit. We find that in this limit the LPM effect holds for medium-induced two-photon ladder emission.

  15. How periodic orbit bifurcations drive multiphoton ionization

    Huang, S.; Chandre, C; Uzer, T.

    2006-01-01

    The multiphoton ionization of hydrogen by a strong bichromatic microwave field is a complex process prototypical for atomic control research. Periodic orbit analysis captures this complexity: Through the stability of periodic orbits we can match qualitatively the variation of experimental ionization rates with a control parameter, the relative phase between the two modes of the field. Moreover, an empirical formula reproduces quantum simulations to a high degree of accuracy. This quantitative...

  16. Medium-induced multi-photon radiation

    Ma Hao; Salgado, Carlos A [Departamento de Fisica de PartIculas, Universidade de Santiago de Compostela, E-15782 Santiago de Compostela (Spain); Tywoniuk, Konrad [Lund University (Sweden)

    2011-01-01

    We study the spectrum of multi-photon radiation off a fast quark in medium in the BDMPS/ASW approach. We reproduce the medium-induced one-photon radiation spectrum in dipole approximation, and go on to calculate the two-photon radiation in the Moliere limit. We find that in this limit the LPM effect holds for medium-induced two-photon ladder emission.

  17. Fundamental studies of molecular multiphoton ionization

    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

  18. First multiphoton tomography of brain in man

    König, Karsten; Kantelhardt, Sven R.; Kalasauskas, Darius; Kim, Ella; Giese, Alf

    2016-03-01

    We report on the first two-photon in vivo brain tissue imaging study in man. High resolution in vivo histology by multiphoton tomography (MPT) including two-photon FLIM was performed in the operation theatre during neurosurgery to evaluate the feasibility to detect label-free tumor borders with subcellular resolution. This feasibility study demonstrates, that MPT has the potential to identify tumor borders on a cellular level in nearly real-time.

  19. Statistical analysis on activation and photo-bleaching of step-wise multi-photon activation fluorescence of melanin

    Gu, Zetong; Lai, Zhenhua; Zhang, Xi; Yin, Jihao; DiMarzio, Charles A.

    2015-03-01

    Melanin is regarded as the most enigmatic pigments/biopolymers found in most organisms. We have shown previously that melanin goes through a step-wise multi-photon absorption process after the fluorescence has been activated with high laser intensity. No melanin step-wise multi-photon activation fluorescence (SMPAF) can be obtained without the activation process. The step-wise multi-photon activation fluorescence has been observed to require less laser power than what would be expected from a non-linear optical process. In this paper, we examined the power dependence of the activation process of melanin SMPAF at 830nm and 920nm wavelengths. We have conducted research using varying the laser power to activate the melanin in a point-scanning mode for multi-photon microscopy. We recorded the fluorescence signals and position. A sequence of experiments indicates the relationship of activation to power, energy and time so that we can optimize the power level. Also we explored regional analysis of melanin to study the spatial relationship in SMPAF and define three types of regions which exhibit differences in the activation process.

  20. Multiphoton imaging to distinguish grana and starch inside an intact leaf

    Chen, Mei-Yu; Zhuo, Guan-Yu; Chen, Po-Fu; Wu, Pei-Chun; Liu, Tzu-Ming; Chu, Shi-Wei

    2013-02-01

    We have demonstrated a straightforward and noninvasive method to identify the distribution of grana and starch within an intact leaf. Grana and starch are the major functional structures for photosynthesis and energy storage of plant, respectively. Both exhibit highly ordered molecular structures and appear as micrometer-sized granules inside chloroplasts. In order to distinguish grana and starch, we used multiphoton microscopy, with simultaneous acquisition of two photon fluorescence (2PF) and second harmonic generation (SHG) signals. Consequently, SHG is found on both grana and starch while 2PF from chlorophyll indicates the identity of grana.

  1. The multiphoton ionization of uranium hexafluoride

    Multiphoton ionization (MPI) time-of-flight mass spectroscopy and photoelectron spectroscopy studies of UF6 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 UFx+ fragment ions. The laser power dependence of Un+ ion signals indicates that saturation can occur for many of the steps required for their ionization. The doubly-charged uranium ion (U2+) intensity is much greater than that of the singly-charged uranium ion (U+). For the case of the tunable dye laser experiments, the Un+ (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 U2+ ion and the absence or very small intensities of UFx+ fragments, along with the unstructured 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. The data argue against step-wise photodissociation of UFx+ (x = 5,6) ions. Neither the neutral ladder nor the ion ladder mechanisms adequately describe the ionization phenomena observed. These results suggest an alternate mechanism which better explains the multiphoton excitation and dissociative ionization of UF6. It is likely that the multiphoton excitation of UF6 under these experimental conditions results in a superexcited molecule, UF6**, which primarily dissociates into Un+ (through multiple channels), fluorine atoms, and slow electrons. The excitation of such superexcited molecules may be facilitated by the existence of a previously reported giant resonance at 12-14 eV

  2. Intense-field multiphoton ionization of helium

    The dynamics of multiphoton ionization of helium are investigated through numerical integration of the two-electron time-dependent Schroedinger equation. Using this work as a benchmark, a new single-active-electron model is introduced that gives agreement with He ionization rates to within a few per cent on average, and gives good agreement with He harmonic generation spectra over a laser intensity range of 0.5x1014 to 8.0x1014 W cm-2, and frequencies corresponding to four- and five-photon ionization. (author). Letter-to-the-editor

  3. Multiphoton imaging the disruptive nature of sulfur mustard lesions

    Werrlein, Robert J.; Braue, Catherine R.; Dillman, James F.

    2005-03-01

    Sulfur mustard [bis-2-chloroethyl sulfide] is a vesicating agent first used as a weapon of war in WWI. It causes debilitating blisters at the epidermal-dermal junction and involves molecules that are also disrupted by junctional epidermolysis bullosa (JEB) and other blistering skin diseases. Despite its recurring use in global conflicts, there is still no completely effective treatment. We have shown by imaging human keratinocytes in cell culture and in intact epidermal tissues that the basal cells of skin contain well-organized molecules (keratins K5/K14, α6β4 integrin, laminin 5 and α3β1 integrin) that are early targets of sulfur mustard. Disruption and collapse of these molecules is coincident with nuclear displacement, loss of functional asymmetry, and loss of polarized mobility. The progression of this pathology precedes basal cell detachment by 8-24 h, a time equivalent to the "clinical latent phase" that defines the extant period between agent exposure and vesication. Our images indicate that disruption of adhesion-complex molecules also impairs cytoskeletal proteins and the integration of structures required for signal transduction and tissue repair. We have recently developed an optical system to test this hypothesis, i.e., to determine whether and how the early disruption of target molecules alters signal transduction. This environmentally controlled on-line system provides a nexus for real-time correlation of imaged lesions with DNA microarray analysis, and for using multiphoton microscopy to facilitate development of more effective treatment strategies.

  4. Signatures of tunneling and multiphoton ionization by short-laser pulses: The partial-wave distribution

    Arbó, Diego G.; Lemell, Christoph; Burgdörfer, Joachim

    2015-09-01

    We analyze the two-dimensional angular momentum-energy distribution of electrons emitted from argon by short laser pulses. We identify characteristic features of both multiphoton and tunneling ionization in the partial-wave distribution for Keldysh parameters close to unity. We observe a remarkable degree of quantum-classical correspondence in the photoinization process which becomes even more pronounced after intensity averaging over the focal volume. We derive an energy-dependent cut-off for the highest angular momentum accessible within the framework of the strong-field approximation, which accurately reproduces the partial wave distributions found from solutions of the time-dependent Schrödinger equation.

  5. The infrared multiphoton dissociation of three nitrolkanes

    Wodtke, A. M.; Hintsa, E. J.; Lee, Y. T.

    1986-01-01

    Infrared multiphoton dissociation in a molecular beam has been studied in order to elucidate the collision free, 'thermal' chemistry and dynamics of nitromethane, nitroethane and 2-nitropropane. The isomerization of CH3NO2 to CH3ONO was observed by detecting the CH3O and NO products from the dissociation of the very internally hot, isomerized nitromethane. A novel application of RRKM theory was used to estimate the barrier height to isomerization at 55.5 kcal/mol. The barrier height determination method was tested and found to give excellent results by applying it to the determintaion of the barrier height to HONO elimination from nitroethane, a value which is well known from activation energy measurements. The method was then applied to the case of HONO elimination from 2-nitropropane and it appears that there is good to believe that the barrier height is 3-5 kcal/mol lower in 2-nitropropane than in nitroethane. The success of this method for determining barrier heights shows how a microscopic molecular beam experiment, using infrared multiphoton dissociation where the concept of temperature has no place, can be quantitatively related to pyrolysis experiments which are conducted under collisional, thermal conditions and measure phenomenological quantities such as activation energies.

  6. Single- and multiphoton ionization processes in molecules

    This dissertation is theoretical in nature and can be separated into two main areas: (1) single- and multiphoton ionization studies of a novel photoelectron effect, and (2) single-photon ionization studies of simple clusters as models for adsorbate photoemission. The first area centers on the phenomenon of circular dichroism in photoelectron angular distributions (CDAD). CDAD is shown to exist from oriented linear molecules, adsorbed atoms, and aligned atoms and molecules in the gas phase. The calculations presented here are the first to demonstrate the experimental feasability of CDAD studies. CDAD is shown to be a measureable effect which exists because the photoelectron collection direction can break the symmetry of these otherwise highly symmetric systems. As a direct results of the work presented here, CDAD has now been observed experimentally. Coupled with resonantly enhanced multiphoton ionization (REMPI), CDAD is shown to be a powerful probe of unknown alignment in gas phase atomic and molecular samples. The second area of research focuses on the simple oriented molecules NiCO and NiN2 as models for the corresponding adsorbate systems. These simple models provide insight into features observed in the experimental angle-resolved photoemission spectra

  7. Multiphoton ionization of large water clusters

    Water clusters are multimers of water molecules held together by hydrogen bonds. In the present work, multiphoton ionization in the UV range coupled with time of flight mass spectrometry has been applied to water clusters with up to 160 molecules in order to obtain information on the electronic states of clusters of different sizes up to dimensions that can approximate the bulk phase. The dependence of ion intensities of water clusters and their metastable fragments produced by laser ionization at 355 nm on laser power density indicates a (3+1)-photon resonance-enhanced multiphoton ionization process. It also explains the large increase of ionization efficiency at 355 nm compared to that at 266 nm. Indeed, it was found, by applying both nanosecond and picosecond laser ionization with the two different UV wavelengths, that no water cluster sequences after n = 9 could be observed at 266 nm, whereas water clusters up to m/z 2000 Th in reflectron mode and m/z 3000 Th in linear mode were detected at 355 nm. The agreement between our findings on clusters of water, especially true in the range with n > 10, and reported data for liquid water supports the hypothesis that clusters above a critical dimension can approximate the liquid phase. It should thus be possible to study clusters just above 10 water molecules, for getting information on the bulk phase structure

  8. Multiphoton imaging of quantum dot bioconjugates in cultured cells following Nd:YLF laser excitation

    Serrano, Elba E.; Knight, V. B.

    2005-04-01

    Quantum dot bioconjugates offer unprecedented opportunities for monitoring biological processes and molecular interactions in cells, tissues, and organs. We are interested in developing applications that permit investigation of physiological processes and cytoskeletal organization in live cells, and allow imaging of complex organs, such as the auditory and vestibular sensory structures of the inner ear. Multiphoton microscopy is a powerful technique for acquiring images from deep within a sample while reducing phototoxic effects of laser light exposure on cells. Previous studies have established that a solid-state Nd:YLF laser can be used to acquire two-photon and three-photon images from live cells while minimizing phototoxic side effects (Wokosin et al., 1996, Bioimaging, 4:208-214; Squirrell et al., 1999, Nature Biotechnology, 8:763-767). We present here the results of experiments using an all-solid-state Nd:YLF 1047 nm femtosecond laser (Microlase DPM1000) source to excite quantum dot bioconjugates. Cells were labeled with Qdot (Quantum Dot Corporation) bioconjugates or with Alexa Fluor (Molecular Probes) bioconjugates and then imaged with a BioRad 1024 confocal microscope configured for multiphoton imaging using internal or external (non-descanned) detectors. Results demonstrate that the Nd:YLF laser can be used to stimulate fluorescence emission of quantum dots and Alexa Fluor bioconjugates in cultured amphibian (Xenopus) and mammalian (rat, chinese hamster) cells. We conclude that the Nd:YLF laser is a viable excitation source that extends the applicability of quantum dots for investigation of biological processes using multiphoton microscopy.

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

    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.

  10. R-matrix Floquet theory of molecular multiphoton processes: II. Multiphoton ionization of H2

    Multiphoton ionization rates for H2 immersed in an intense linearly polarized laser field are calculated using the recently developed R-matrix Floquet theory of molecular multiphoton processes. We assume that the H2 molecule is aligned along the laser polarization direction and we adopt the fixed-nuclei approximation, in which the motion of the target electrons is calculated in the laser field and in the field of the nuclei, which are assumed to be fixed in space. An accurate multi-state wavefunction is employed to calculate one-, two- and four-photon ionization rates for H2 at several internuclear separations over a range of frequencies and intensities. Analysis of the ionization rates reveals the important role played both by resonances corresponding to Rydberg bound states converging to the H2+ ion ground state and by doubly excited states converging to the H2+ ion first excited state. These resonances give rise to resonant enhanced multiphoton ionization peaks in many of the ionization rates studied in this paper, and their possible role in controlling the vibrational population of the final H2+ ion is discussed. (author)

  11. In vivo multiphoton tomography of inflammatory tissue and melanoma

    Riemann, Iris; Dimitrow, Enrico; Kaatz, Martin; Fluhr, Joachim; Elsner, Peter; Kobow, Jens; Konig, Karsten

    2005-04-01

    Multiphoton optical tomography provides the capability of non-invasive optical sectioning of skin with high spatial and intracellular resolution as well as high NIR (near infrared) light penetration into pigmented skin areas. The imaging system DermaInspect based on femtosecond laser pulses was used to perform multiphoton optical tomography in clinical studies. Patients with abnormal pigmented tissues were imaged in vivo. After the multiphoton imaging procedure, biopsies were taken, imaged again and further processed with standard histological methods. We report on preliminary results. The visualization of pigmented cell clusters based on non-linear luminescence using the novel multiphoton device was possible. These clusters could be clearly distinguished from non-pigmented cells. Cancerous tissues showed significant differences in the cell structure of the epidermal layers. The system DermaInspect might become a high resolution diagnostic tool for melanoma diagnostics.

  12. Multi-photon entanglement in high dimensions

    Malik, Mehul; Huber, Marcus; Krenn, Mario; Fickler, Robert; Zeilinger, Anton

    2015-01-01

    Entanglement lies at the heart of quantum mechanics $-$ as a fundamental tool for testing its deep rift with classical physics, while also providing a key resource for quantum technologies such as quantum computation and cryptography. In 1987 Greenberger, Horne, and Zeilinger realized that the entanglement of more than two particles implies a non-statistical conflict between local realism and quantum mechanics. The resulting predictions were experimentally confirmed by entangling three photons in their polarization. Experimental efforts since have singularly focused on increasing the number of particles entangled, while remaining in a two-dimensional space for each particle. Here we show the experimental generation of the first multi-photon entangled state where both $-$ the number of particles and the number of dimensions $-$ are greater than two. Interestingly, our state exhibits an asymmetric entanglement structure that is only possible when one considers multi-particle entangled states in high dimensions....

  13. Multi-photon entanglement in high dimensions

    Malik, Mehul; Erhard, Manuel; Huber, Marcus; Krenn, Mario; Fickler, Robert; Zeilinger, Anton

    2016-04-01

    Forming the backbone of quantum technologies today, entanglement has been demonstrated in physical systems as diverse as photons, ions and superconducting circuits. Although steadily pushing the boundary of the number of particles entangled, these experiments have remained in a two-dimensional space for each particle. Here we show the experimental generation of the first multi-photon entangled state where both the number of particles and dimensions are greater than two. Two photons in our state reside in a three-dimensional space, whereas the third lives in two dimensions. This asymmetric entanglement structure only appears in multiparticle entangled states with d > 2. Our method relies on combining two pairs of photons, high-dimensionally entangled in their orbital angular momentum. In addition, we show how this state enables a new type of ‘layered’ quantum communication protocol. Entangled states such as these serve as a manifestation of the complex dance of correlations that can exist within quantum mechanics.

  14. Lippmann-Schwinger description of multiphoton ionization

    Ivanov, I A

    2005-01-01

    We outline a formalism and develop a computational procedure to treat the process of multiphoton ionization (MPI) of atomic targets in strong laser fields. We treat the MPI process nonperturbatively as a decay phenomenon by solving a coupled set of the integral Lippmann-Schwinger equations. As basic building blocks of the theory we use a complete set of field-free atomic states, discrete and continuous. This approach should enable us to provide both the total and differential cross-sections of MPI of atoms with one or two electrons. As an illustration, we apply the proposed procedure to a simple model of MPI from a square well potential and to the hydrogen atom.

  15. Multiphoton dissociative ionization of molecular deuterium

    The kinetic energy spectra of deuterium ions produced from D2 arising from collision-free subpicosecond irradiation at 248 nm with intensities spanning the 10/sup 13/--10/sup 16/-W/cm2 range have been measured by time-of-flight analysis. The behaviors of the kinetic energy distributions of the fragments and the relative abundances of atomic (D+) and molecular (D2+) ions reveal the presence of two mechanisms of multiphoton dissociative ionization. Calibration of the energy scale for D+ is facilitated by comparison with He/sup 2+/. For intensities in the 10/sup 13/--10/sup 15/-W/cm2 region, intermediate three-photon resonances and the optical Stark shift play important roles. At an intensity /similar to/0/sup 16/ W/cm2, a direct transition from the molecular ground state to the dissociative ionic level appears as a significant channel. No evidence of direct double ionization was observed

  16. Soliton dynamics in the multiphoton plasma regime

    Husko, Chad A; Colman, Pierre; Zheng, Jiangjun; De Rossi, Alfredo; Wong, Chee Wei; 10.1038/srep01100

    2013-01-01

    Solitary waves have consistently captured the imagination of scientists, ranging from fundamental breakthroughs in spectroscopy and metrology enabled by supercontinuum light, to gap solitons for dispersionless slow-light, and discrete spatial solitons in lattices, amongst others. Recent progress in strong-field atomic physics include impressive demonstrations of attosecond pulses and high-harmonic generation via photoionization of free-electrons in gases at extreme intensities of 1014 Wcm2. Here we report the first phase-resolved observations of femtosecond optical solitons in a semiconductor microchip, with multiphoton ionization at picojoule energies and 1010 Wcm2 intensities. The dramatic nonlinearity leads to picojoule observations of free-electron-induced blue-shift at 1016 cm3 carrier densities and self-chirped femtosecond soliton acceleration. Furthermore, we evidence the time-gated dynamics of soliton splitting on-chip, and the suppression of soliton recurrence due to fast free-electron dynamics. Thes...

  17. Advances in multi-photon processes and spectroscopy, v.3

    Lin, Sheng H

    1987-01-01

    In view of the rapid growth in both experimental and theoretical studies of multiphoton processes and multiphoton spectroscopy of atoms, ions, and molecules in chemistry, physics, biology and material sciences, this series contains review papers exploring the growth. They are readable not only by active researchers in these areas but also by those who are not experts in the field but intend to enter the field.

  18. Multiphoton above threshold effects in strong-field fragmentation

    Madsen, C. B.; Anis, F; Madsen, L. B.; Esry, B. D.

    2012-01-01

    We present a study of multiphoton dissociative ionization from molecules. By solving the time-dependent Schr\\"{o}dinger equation for H$_2^+$ 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 the energies of all fragments are accounted for. Our current work provides a new avenue to analyze strong-field fragmentation that l...

  19. Advances in multi-photon processes and spectroscopy, v.2

    Lin, Sheng H

    1986-01-01

    This volume focuses on the recent rapid growth in both experimental and theoretical studies of multiphoton processes and multiphoton spectroscopy of atoms, ions and molecules in chemistry, physics, biology, material sciences, It contains papers readable by active researchers and by those who intend to enter it. Theory and experiment are equally emphasized, and each review article is written in a self-contained manner by experts in the field so that readers learn the subject without much preparation.

  20. Molecular detection by multiphoton photoionization: Final report, 1 November 1985-1 November 1986

    This paper discusses the use of multiphoton ionization techniques to detection molecules at a sensitivity approaching one-molecule/cm3. Multiphoton fragmentation processes and ion spectroscopy are discussed. 10 refs., 3 figs

  1. Multiphoton photochemistry of red fluorescent proteins in solution and live cells.

    Drobizhev, Mikhail; Stoltzfus, Caleb; Topol, Igor; Collins, Jack; Wicks, Geoffrey; Mikhaylov, Alexander; Barnett, Lauren; Hughes, Thomas E; Rebane, Aleksander

    2014-08-01

    Genetically encoded fluorescent proteins (FPs), and biosensors based on them, provide new insights into how living cells and tissues function. Ultimately, the goal of the bioimaging community is to use these probes deep in tissues and even in entire organisms, and this will require two-photon laser scanning microscopy (TPLSM), with its greater tissue penetration, lower autofluorescence background, and minimum photodamage in the out-of-focus volume. However, the extremely high instantaneous light intensities of femtosecond pulses in the focal volume dramatically increase the probability of further stepwise resonant photon absorption, leading to highly excited, ionizable and reactive states, often resulting in fast bleaching of fluorescent proteins in TPLSM. Here, we show that the femtosecond multiphoton excitation of red FPs (DsRed2 and mFruits), both in solution and live cells, results in a chain of consecutive, partially reversible reactions, with individual rates driven by a high-order (3-5 photon) absorption. The first step of this process corresponds to a three- (DsRed2) or four-photon (mFruits) induced fast isomerization of the chromophore, yielding intermediate fluorescent forms, which then subsequently transform into nonfluorescent products. Our experimental data and model calculations are consistent with a mechanism in which ultrafast electron transfer from the chromophore to a neighboring positively charged amino acid residue triggers the first step of multiphoton chromophore transformations in DsRed2 and mFruits, consisting of decarboxylation of a nearby deprotonated glutamic acid residue. PMID:25004113

  2. High-resolution multiphoton optical tomography of tissues: an in vitro and in vivo study

    Riemann, Iris; Schenke-Layland, Katja; Ehlers, Alexander; Dimitrow, Enrico; Kaatz, Martin; Elsner, Peter; Martin, Sven; König, Karsten

    2006-03-01

    Multiphoton optical tomography based on NIR (near-infrared) femtosecond laser pulses provides non-invasive optical sectioning of skin with high spatial intracellular resolution and high tissue penetration. The imaging system DermaInspect was used to perform this technology in clinical studies in vivo on patients with suspicious melanoma. Pigmented cell clusters based on non-linear luminescence were clearly distinguished from non-pigmented cells in the epidermis using the autofluorescence of endogenous fluorophores like NAD(P)H, flavins, keratin, elastin, collagen and melanin. Some of the investigated tissues showed differences in the structure of the epidermal layers and the presence of dendritic cells compared to normal skin. Multiphoton laser microscopy was used to visualize extracellular matrix (ECM) structures of native and tissueengineered heart valves. The quality of the resulting 3-D images allowed an exact differentiation between collagenous and elastic fibers. The analysis of heart valve tissues of patients with cardiomyopathy revealed a dramatic loss of its capability to generate SH (second harmonic), indicating a structural deformation of the collagenous fibers, which was virtually impossible to obtain by routine histological or immunohistological staining. These results indicate that NIR femtosecond laser scanning systems can be employed as novel non-invasive optical technology for 3-D resolved ECM component imaging and in vitro and in vivo tissue diagnosis.

  3. High-resolution multimodal clinical multiphoton tomography of skin

    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.

  4. The multiphoton ionization of uranium hexafluoride

    Multiphoton ionization (MPI) time-of-flight mass spectroscopy and photoelectron spectroscopy studies of UF6 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 UFx+ fragment ions even at the lowest laser power densities at which signal could be detected. The laser power dependence of Un+ ions signals indicates that saturation can occur for many of the steps required for their ionization. In general, the doubly-charged uranium ion (U2+) intensity is much greater than that of the singly-charged uranium ion (U+). For the case of the tunable dye laser experiments, the Un+ (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 U2+ ion and the absence or very small intensities of UFx+ 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

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

    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.)

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

    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.)

  7. Preparation of metallo-dielectric photonic crystals by multi-photon direct laser writing

    Kuebler, Stephen M.; Tal, Amir; Chen, Yun-Sheng

    2008-02-01

    Metallo-dielectric photonic crystals (MDPCs) can exhibit intriguing and potentially useful optical properties, including ultra-wide photonic bandgaps, engineered thermal emission, and negative refractive index. But access to such materials has been limited by the lack of suitable methods for their preparation. We have developed a route to three-dimensional (3D) MDPCs that involves fabricating a polymeric pre-form by multi-photon direct laser writing and then conformally depositing metal onto the pre-form by electroless metallization. We use the approach to prepare silver- and copper-plated "woodpile" PCs having face-centered tetragonal symmetry and unit-cell period of several micrometers. The resulting 3D metallized structures exhibit mid-infrared reflectance that is consistent with theory and experimental observations obtained for MDPCs prepared by other routes. These data indicate that multi-photon direct laser writing coupled with electroless metallization is a viable route to complex 3D MDPCs of many symmetries and basis sets and provides a path for integrating such structures with other micron-scale optical elements.

  8. Optical Investigation of the Intergrowth Structure and Accessibility of Brønsted Acid Sites in Etched SSZ-13 Zeolite Crystals by Confocal Fluorescence Microscopy

    Sommer, L.; Svelle, S.; Lillerud, K.-P.; Stöcker, M; Weckhuysen, B.M.; Olsbye, U.

    2013-01-01

    Template decomposition followed by confocal fluorescence microscopy reveals a tetragonal-pyramidal intergrowth of subunits in micrometer-sized nearly cubic SSZ-13 zeolite crystals. In order to accentuate intergrowth boundaries and defect-rich areas within the individual large zeolite crystals, a tre

  9. Selective enhancement of resonant multiphoton ionization with strong laser fields

    Li, Min; Luo, Siqiang; Zhou, Yueming; Zhang, Qingbin; Lan, Pengfei; Lu, Peixiang

    2015-01-01

    High-resolution photoelectron momentum distributions of Xe atom ionized by 800-nm linearly polarized laser fields have been traced at intensities from 1.1*1013 W/cm2 to 3.5*1013 W/cm2 using velocity-map imaging techniques. At certain laser intensities, the momentum spectrum exhibits a distinct double-ring structure for low-order above-threshold ionization, which appears to be absent at lower or higher laser intensities. By investigating intensity-resolved photoelectron energy spectrum, we find that this double-ring structure originates from resonant multiphoton ionization involving multiple Rydberg states of atoms. Varying the laser intensity, we can selectively enhance multiphoton excitation of atomic Rydberg populations. The photoelectron angular distributions of multiphoton resonance are also investigated for the low-order above threshold ionization.

  10. Dynamic resonances and tunnelling in the multiphoton ionization of argon

    We present results of wavepacket simulations for multiphoton ionization in argon. A single active electron model is applied to estimate the single-electron ionization rates and photoelectron energy distributions for λ = 390 nm light with intensities up to I = 2 x 1014 W cm-2. The multiphoton ionization rates are compared with R-matrix Floquet calculations and found to be in very good agreement. The photoelectron energy distribution is used to study the nature of ionization at the higher intensities. Our results are consistent with recent calculations and experiments which show the imprint of the tunnelling process in the multiphoton regime. For few-cycle intense pulses, we find that the strong modulation of intensity and increased bandwidth leads to dynamic mixing of the 3d and 5s resonances.

  11. Advances in multi-photon processes and spectroscopy, v.5

    Lin, Sheng H

    1989-01-01

    In view of the rapid growth in both experimental and theoretical studies of multiphoton processes and multiphoton spectroscopy of atoms, ions and molecules in many interdisciplinary fields, an Advanced Series that contains review papers readable not only to active researchers in these areas but also to those who are not experts in the field but intend to enter the field is very necessary. This series attempts to serve this purpose. Each review article is written in a self-contained manner by the experts in the area so that the readers can grasp the knowledge in the area without too much prepar

  12. Unambiguous atomic Bell measurement assisted by multiphoton states

    Torres, Juan Mauricio; Bernád, József Zsolt; Alber, Gernot

    2016-05-01

    We propose and theoretically investigate an unambiguous Bell measurement of atomic qubits assisted by multiphoton states. The atoms interact resonantly with the electromagnetic field inside two spatially separated optical cavities in a Ramsey-type interaction sequence. The qubit states are postselected by measuring the photonic states inside the resonators. We show that if one is able to project the photonic field onto two coherent states on opposite sites of phase space, an unambiguous Bell measurement can be implemented. Thus, our proposal may provide a core element for future components of quantum information technology such as a quantum repeater based on coherent multiphoton states, atomic qubits and matter-field interaction.

  13. Quantum cryptography with entangled multiphotons of the same polarization

    Multiphoton entanglement in the same polarization has been theoretically shown to be obtainable by type-I spontaneous parametric down-conversion (SPDC), which can generate bright pulses more easily than type-II SPDC. A quantum cryptographic protocol utilizing photon number coding of the detected type-I entangled multiphotons is described. The information capacity versus photon number is calculated after considering the transmission loss inside the optical fiber and the detector efficiency, considering possible attacks to the scheme. The result thus achieved compares very favorably with all other schemes employing entanglement

  14. Effect of multiphoton ionization on performance of crystalline lens.

    Gupta, Pradeep Kumar; Singh, Ram Kishor; Strickland, D; Campbell, M C W; Sharma, R P

    2014-12-15

    This Letter presents a model for propagation of a laser pulse in a human crystalline lens. The model contains a transverse beam diffraction effect, laser-induced optical breakdown for the creation of plasma via a multiphoton ionization process, and the gradient index (GRIN) structure. Plasma introduces the nonlinearity in the crystalline lens which affects the propagation of the beam. The multiphoton ionization process generates plasma that changes the refractive index and hence leads to the defocusing of the laser beam. The Letter also points out the relevance of the present investigation to cavitation bubble formation for restoring the elasticity of the eyes. PMID:25502994

  15. Observation of detection-dependent multi-photon coherence times

    Ra, Young-Sik; Tichy, Malte C.; Lim, Hyang-Tag;

    2013-01-01

    which the multi-photon coherence time, defined by the width of the interference signal, depends on the number of interfering photons and on the measurement scheme chosen to detect the particles. A theoretical analysis reveals that all multi-photon interferences with more than two particles feature this...... dependence, which can be attributed to higher-order effects in the mutual indistinguishability of the particles. As a striking consequence, a single, well-defined many-particle quantum state can exhibit qualitatively different degrees of interference, depending on the chosen observable. Therefore, optimal...... sensitivity in many-particle quantum interferometry can only be achieved by choosing a suitable detection scheme....

  16. Quantitative studies of multiphoton ionization using tunable VUV radiation

    The storage ring free electron laser makes studies of multiphoton ionization in the vacuum ultraviolet possible. At relatively low laser intensities one can study two-photon resonant three-photon ionization of atoms in a regime where perturbation theory works well. In this regime cross sections for the multiphoton processes can be measured accurately and then used for sensitive, quantitative detection of atoms. At higher intensities higher-order processes such as multiple ionization can take place. The tunability, variable pulse length, and well characterized spatial distribution of the FEL is important in unraveling the mechanisms of these processes

  17. Nanosecond Versus, Picosecond Molecular Multiphoton Fragmentation of Ketene and Cyclohexane

    Castillejo, M.; Martín, M; de Nalda, R.

    1998-01-01

    Multiphoton dissociation of ketene was studied at 290.5 nm after irradiation of the first excited singlet state with two different laser pulsewidths of 4 ns and 10 ps. Analysis of the photofragment fluorescence spectra in the two time domains sheds light on the mechanisms that are responsible for the production of the observed CH(A2Δ),  (B2Σ−) and C2(d3Πg) photofragments. Multiphoton ionization of cyclohexane induced by a visible laser at 581 nm and its second harmonic at 290.5 nm was measure...

  18. Multiphoton above threshold effects in strong-field fragmentation

    Madsen, C B; Madsen, L B; Esry, B D

    2012-01-01

    We present a study of multiphoton dissociative ionization from molecules. By solving the time-dependent Schr\\"{o}dinger equation for H$_2^+$ 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 the energies of all fragments are accounted for. Our current work provides a new avenue to analyze strong-field fragmentation that leads to a deeper understanding of the correlated molecular dynamics.

  19. Multiphoton above threshold effects in strong-field fragmentation

    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....

  20. Robert Feulgen Prize Lecture. Laser tweezers and multiphoton microscopes in life sciences.

    König, K

    2000-08-01

    Near infrared (NIR) laser microscopy enables optical micromanipulation, piconewton force determination, and sensitive fluorescence studies by laser tweezers. Otherwise, fluorescence images with high spatial and temporal resolution of living cells and tissues can be obtained via non-resonant fluorophore excitation with multiphoton NIR laser scanning microscopes. Furthermore, NIR femtosecond laser pulses at TW/cm2 intensities can be used to realize non-invasive contact-free surgery of nanometer-sized structures within living cells and tissues. Applications of these novel versatile NIR laser-based tools for the determination of motility forces, coenzyme and chlorophyll imaging, three-dimensional multigene detection, non-invasive optical sectioning of tissues ("optical biopsy"), functional protein imaging, and nanosurgery of chromosomes are described. PMID:11052257

  1. Generation of multiphoton entangled quantum states by means of integrated frequency combs.

    Reimer, Christian; Kues, Michael; Roztocki, Piotr; Wetzel, Benjamin; Grazioso, Fabio; Little, Brent E; Chu, Sai T; Johnston, Tudor; Bromberg, Yaron; Caspani, Lucia; Moss, David J; Morandotti, Roberto

    2016-03-11

    Complex optical photon states with entanglement shared among several modes are critical to improving our fundamental understanding of quantum mechanics and have applications for quantum information processing, imaging, and microscopy. We demonstrate that optical integrated Kerr frequency combs can be used to generate several bi- and multiphoton entangled qubits, with direct applications for quantum communication and computation. Our method is compatible with contemporary fiber and quantum memory infrastructures and with chip-scale semiconductor technology, enabling compact, low-cost, and scalable implementations. The exploitation of integrated Kerr frequency combs, with their ability to generate multiple, customizable, and complex quantum states, can provide a scalable, practical, and compact platform for quantum technologies. PMID:26965623

  2. Multiphoton Coherent Manipulation in Large Spin Qubits

    Chiorescu, Irinel

    2009-03-01

    Manipulation of quantum information allows certain algorithms to be performed at unparalleled speeds. Photons are an ideal choice to manipulate qubits as they interact with quantum systems in predictable ways. They are a versatile tool for manipulating, reading/coupling qubits and for encoding/transferring quantum information over long distances. Spin-based qubits have well known behavior under photon driving and can be potentially operated up to room temperature. When diluted enough to avoid uncontrolled spin-spin interactions, a variety of spin qubits show long coherence times, e.g. the nitrogen vacancies in pure diamonds (1,2), nitrogen atoms trapped in a C60 cage (3), Ho3+ and Cr5+ ions (4,5) and molecular magnets (6,7). We have used large spin Mn2+ ions (S=5/2) to realize a six level system that can be operated by means of single as well as multi-photon coherent Rabi oscillations (8). This spin system has a very small anisotropy whose effect can be tuned in-situ to turn the system into a multi-level harmonic system. This offer new ways of manipulating, reading and resetting a spin qubit. Decoherence effects are strongly reduced by the quasi-isotropic electron interaction with the crystal field and with the 55Mn nuclear spins. [0pt] 1. R. Hanson et al., Science 320, 352 (2008). [0pt] 2. M.V. Gurudev Dutt et al., Science 316, 1312 (2007). [0pt] 3. G.W. Morley et al., Phys. Rev. Lett. 98, 220501 (2007). [0pt] 4. S. Bertaina et al., Nat. Nanotech. 2, 39 (2007). [0pt] 5. S. Nellutla et al., Phys. Rev. Lett. 99, 137601 (2007). [0pt] 6. A. Ardavan et al., Phys. Rev. Lett. 98, 057201 (2007). [0pt] 7. S. Bertaina et al., Nature 453, 203,(2008). [0pt] 8. S. Bertaina et al., submitted.

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

    Woutersen, S.; Milan,, M; Lange; 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 1D excited state, prepared by in situ photodissociation of H2S. The observed states derive from the (2Do)5p and (2Po)4p configurations. For the (2Do)5p 3F and (2Po)4p 3D triplets, extensive photoele...

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

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

    1997-05-01

    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 1D excited state, prepared by in situ photodissociation of H2S. The observed states derive from the (2Do)5p and (2Po)4p configurations. For the (2Do)5p 3F and (2Po)4p 3D triplets, extensive photoelectron spectroscopic studies enable a detailed comparison of the autoionization and photoionization rates of these states.

  5. Advances in time-dependent methods for multiphoton processes

    This paper discusses recent theoretical results on above threshold ionization harmonic generation and high-frequency, high intensity suppression of ionization. These studies of multiphoton processes in atoms and molecules for short, intense pulsed optical lasers have been carried out using techniques which involve the explicit solution of the time-dependent Schroedinger equation. 43 refs., 5 figs

  6. Multiphoton ionization of magnesium via an autoionizing state

    N.J. van Druten; R. Trainham; H.G. Muller

    1994-01-01

    Multiphoton single and double ionization of magnesium was studied by measuring electron energy spectra and ion mass spectra using 1-ps laser pulses in the 580-595-nm wavelength and 1012-1013-W/cm2 intensity range. In single ionization the (3p)2 1S doubly excited autoionizing state, resonant at the f

  7. All short pulse multiphoton ionization is resonant ionization

    Energy resolved photoelectron spectra of multiphoton ionization taken with a 300 fs laser pulse at 616 nm shows that the ionization probability is highly structured as a function of laser intensity. The spectrum is consistent with all of the ionization occurring at intensity resonances

  8. Multi-photon Absorption in Optical Pumping of Rubidium

    Xu, Xinyi

    2015-01-01

    In optical pumping of rubidium, a new kind of absorption occurs with a higher amplitude of radio frequency current. From measurement of the corresponding magnetic field value where this absorption occurs, there is a conclusion that it is multi-photon absorption. Both the degeneracy and energy of photons contribute to the intensity.

  9. Multi-photon Rabi model: Generalized parity and its applications

    Quantum multi-photon spin–boson model is considered. We solve an operator Riccati equation associated with that model and present a candidate for a generalized parity operator allowing to transform spin–boson Hamiltonian to a block-diagonal form what indicates an existence of the related symmetry of the model.

  10. On a Jaynes-Cummings type model with multiphoton transitions

    The authors present a quantum electrodynamic model, soluble in the dipole and rotating wave approximation, for a three-level atom interacting with a two-mode resonant radiation field through the multiphoton transition mechanism. Population dynamics and photon statistics in this Jaynes-Cummings type model are examined. (author)

  11. Landau-Dykhne approximation for multiphoton dipole-forbidden transitions

    A two-level system in a monochromatic laser field is considered in the Landau-Dykhne approximation under the violation of dipole selection rules. An analytic expression is obtained for the rate of transitions. The multiphoton and tunneling limits are found

  12. Multi-photon entanglement and applications in quantum information

    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

  13. Multi-photon entanglement and applications in quantum information

    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

  14. Highly resolved intravital striped-illumination microscopy of germinal centers.

    Cseresnyes, Zoltan; Oehme, Laura; Andresen, Volker; Sporbert, Anje; Hauser, Anja E; Niesner, Raluca

    2014-01-01

    Monitoring cellular communication by intravital deep-tissue multi-photon microscopy is the key for understanding the fate of immune cells within thick tissue samples and organs in health and disease. By controlling the scanning pattern in multi-photon microscopy and applying appropriate numerical algorithms, we developed a striped-illumination approach, which enabled us to achieve 3-fold better axial resolution and improved signal-to-noise ratio, i.e. contrast, in more than 100 µm tissue depth within highly scattering tissue of lymphoid organs as compared to standard multi-photon microscopy. The acquisition speed as well as photobleaching and photodamage effects were similar to standard photo-multiplier-based technique, whereas the imaging depth was slightly lower due to the use of field detectors. By using the striped-illumination approach, we are able to observe the dynamics of immune complex deposits on secondary follicular dendritic cells - on the level of a few protein molecules in germinal centers. PMID:24748007

  15. Microscopy with nano focus

    Shaul Aloni

    2005-01-01

    The Handbook of Microscopy for Nanotechnology is recommended for scientists needing to choose which methods to use, but its high price may bar it from students using it as a quick, accessible entré to the topic, says Shaul Aloni.

  16. Random-access Two-photon Microscopy for Neural Activity Observation%用于神经活动观测的随机扫描双光子显微成像

    姜润华; 吕晓华; 李德荣; 全廷伟; 刘秀丽; 骆清铭; 曾绍群

    2012-01-01

    Two-photon microscope has become an important instrument in neuroscience research. However, the current commercial instruments can hardly meet the need for the detection of neural signal in millisecond scale due to their low imaging rates. Fast random-access two-photon microscopy based on acousto-optic deflector (AOD) has the potential for increasing the observation speed while maintaining adequate signal to noise ratio (SNR). We summarize the latest related research progress. It is demonstrated from four parts, including the spatio-temporal evolution theory of the femtosecond laser after passing the angular dispersion devices, dispersion compensation method for AOD, random-access two-photon microscopy instrument, and calcium signal identification method in the instrument applications. In the end, the future development trends for random-access two-photon microscopy are discussed. The systematic and in-deep research on this technology will provide a new tool for the neural activity observation and boost the development of brain science.%双光子荧光显微镜是神经科学研究中的重要观测仪器,但是现有的商品化仪器受限于较低的成像速度,难以满足脑功能研究中毫秒量级神经信号检测的需要.基于声光偏转器的快速随机扫描双光子显微成像技术,有望在保持信噪比的同时提高观测速度.本文综述了这一研究的最新进展,从飞秒激光经过角色散器件后的时空演化理论、声光偏转器的色散补偿方法、随机扫描成像仪器及仪器应用到神经成像时钙信号的识别方法四个方面分别进行介绍,最后分析了随机扫描双光子显微成像技术的发展趋势.这项技术的系统深入研究将为神经活动观测提供一种全新的方法,推动脑科学研究的发展.

  17. Multiphoton imaging of excised normal skin and keloid scar: preliminary investigations

    Brewer, Michael B.; Yeh, Alvin T.; Torkian, Behrooz; Sun, Chung-Ho; Tromberg, Bruce J.; Wong, Brian J.

    2004-07-01

    Wound healing is a physiologic process that acts to repair disruptions in the continuity of tissue caused by injury or surgical incision. Keloids and hypertrophic scars are forms of aberrant wound healing, which are characterized by the overproduction of collagen, resulting in an excessive amount of scar tissue. Keloid tumors, by definition, grow outside the boundary of the original tissue damage. Multiphoton microscopy (MPM) is an imaging technique which allows imaging of living specimens, without the use of fixation or stains. Images of collagen fibers are produced by the second harmonic signal intensity generated by endogenous fluorescence through excitation by infrared laser light. A postauricular keloid tumor was excised from a patient. The tissue was dissected, and a portion was imaged using MPM. Normal skin tissue was isolated from a patient undergoing a facelift. A portion of this tissue was also dissected and imaged using MPM. MPM images were taken using a 63X water immersion objective lens on a two-photon microscope and a titanium-sapphire laser. Images were taken beginning at the surface of the tissue and moving in at intervals of 200 nm to a final depth of 30 μm. The two-photon images were used to reconstruct three-dimensional representations of the collagen matrix within the tissues, which are readily contrasted. Density of the collagen within each tissue was also ascertained using depth dependant decay of the image intensity. Multiphoton imaging was successfully used to image the collagen matrix of normal skin and a keloid scar, demonstrating differences in their microstructures.

  18. Multiphoton dynamics of qutrits in the ultrastrong coupling regime with a quantized photonic field

    Multiphoton resonant excitation of a three-state quantum system (a qutrit) with a single-mode photonic field is considered in the ultrastrong coupling regime, when the qutrit–photonic field coupling rate is comparable to appreciable fractions of the photon frequency. For ultrastrong couplings, the obtained solutions of the Schrödinger equation that reveal multiphoton Rabi oscillations in qutrits with the interference effects leading to the collapse and revival of atomic excitation probabilities at the direct multiphoton resonant transitions

  19. Hamiltonian models of multiphoton processes and four--photon squeezed states via nonlinear canonical transformations

    De Siena, Silvio; Di Lisi, Antonio; Illuminati, Fabrizio

    2002-01-01

    We introduce nonlinear canonical transformations that yield effective Hamiltonians of multiphoton down conversion processes, and we define the associated non-Gaussian multiphoton squeezed states as the coherent states of the multiphoton Hamiltonians. We study in detail the four-photon processes and the associated non-Gaussian four-photon squeezed states. The realization of squeezing, the behavior of the field statistics, and the structure of the phase space distributions show that these state...

  20. Multiphoton dynamics of qutrits in the ultrastrong coupling regime with a quantized photonic field

    Avetissian, H. K., E-mail: avetissian@ysu.am; Avetissian, A. K.; Mkrtchian, G. F. [Yerevan State University 0025, Center of Strong Fields Physics (Armenia); Kibis, O. V. [Novosibirsk State Technical University, Department of Applied and Theoretical Physics (Russian Federation)

    2015-12-15

    Multiphoton resonant excitation of a three-state quantum system (a qutrit) with a single-mode photonic field is considered in the ultrastrong coupling regime, when the qutrit–photonic field coupling rate is comparable to appreciable fractions of the photon frequency. For ultrastrong couplings, the obtained solutions of the Schrödinger equation that reveal multiphoton Rabi oscillations in qutrits with the interference effects leading to the collapse and revival of atomic excitation probabilities at the direct multiphoton resonant transitions.

  1. Multiphoton Rabi Oscillations of Correlated Electrons in Strong Field Nonsequential Double Ionization

    Qing, Liao; Cheng, Huang; Peixiang, Lu

    2011-01-01

    With quantum calculations, we have investigated the multiphoton nonsequential double ionization of helium atoms in intense laser fields at ultraviolet wavelengths. Very surprisingly, we find a so-far unobserved double-circle structure in the correlated electron momentum spectra. The double-circle structure essentially reveals multiphoton Rabi oscillations of two electrons, which are strongly supported by the oscillating population of a certain doubly excited state and by the oscillating double ionization signals. This two-electron multiphoton Rabi effect provides profound understandings of electronic correlations and complicated multiphoton phenomena and is expected to be a new tool for broad applications, such as quantum coherent control.

  2. Multiphoton Rabi oscillations of correlated electrons in strong-field nonsequential double ionization

    With quantum calculations, we have investigated the multiphoton nonsequential double ionization of helium atoms in intense laser fields at ultraviolet wavelengths. Very surprisingly, we found a so-far unobserved double-circle structure in the correlated electron momentum spectra. The double-circle structure essentially reveals multiphoton Rabi oscillations of two electrons, which are strongly supported by the oscillating population of a certain doubly excited state and by the oscillating double ionization signals. This two-electron multiphoton Rabi effect provides a profound understanding of electronic correlations and complicated multiphoton phenomena and is expected to be a new tool for broad applications, such as quantum coherent control. (paper)

  3. Multiphoton Imaging of Ultrasound Bioeffects in the Murine Brain

    Raymond, Scott; Skoch, Jesse; Bacskai, Brian; Hynynen, Kullervo

    2006-05-01

    The purpose of this study was to demonstrate the feasibility of multiphoton imaging in the murine brain during exposure to ultrasound. Our experimental setup coupled ultrasound through the ventral surface of the mouse while allowing imaging through a cranial window from the dorsal surface. Field attenuation was estimated by scanning the field after insertion of a freshly sacrificed mouse; beam profile and peak position were preserved, suggesting adequate targeting for imaging experiments. C57 mice were imaged with a Biorad multiphoton microscope while being exposed to ultrasound (f = 1.029 MHz, peak pressure ˜ 200 kPa, average power ˜ 0.18 W) with IV injection of Optison. We observed strong vasoconstriction coincident with US and Optison, as well as permeabilization of the blood-brain barrier.

  4. Experimental Resonance Enhanced Multiphoton Ionization (REMPI) studies of small molecules

    Dehmer, J. L.; Dehmer, P. M.; Pratt, S. T.; Ohalloran, M. A.; Tomkins, F. S.

    1987-01-01

    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. An overview of current studies of excited molecular states is given to illustrate the principles and prospects of REMPI.

  5. Microwave multiphoton ionization and excitation of helium Rydberg atoms

    We study experimentally and theoretically the detailed field-amplitude dependence of the multiphoton ionization and excitation probability of highly excited n03S helium atoms in a 9.924-GHz linearly polarized microwave electric field. For ionization, with principal quantum numbers in the range n0=25--32, we use a quasistatic analysis that employs integration of the time-dependent Schroedinger equation using basis states of the static field Hamiltonian. The calculated results are used to interpret the observed ionization threshold structure. For excitation, the results of n03S→n03L, L>2 excitation experiments are explained quantitatively and precisely using a theory of multiphoton resonances. We present maps of quasienergy levels that allow the study of the dynamics of the field-switching transients. These transient effects are analyzed along the lines of standard atomic collision theory and are shown to determine the shape of the observed resonances

  6. Theoretical aspects of multiphoton ionization with many resonant excited states

    The variety of the parameters involved in multi-color multi-step ionization makes it a formidable job to obtain a theoretically comprehensible overview of the process. We examine these parameters of such processes as well as commonly used assumptions in theoretical investigations of multiphoton ionization with many resonantly excited levels. The density matrix formalism is adequate to treat resonant multiphoton ionization when the number of the resonant levels involved is not too large, solving the resonantly coupled states separately beyond the lowest order perturbation theory, while the rest of the states are treated perturbatively. An example of such formalism is given for a four-level system ionized with three lasers each of which resonantly couples the adjacent pairs of the atomic states.

  7. A simple model of multiphoton micromachining in silk hydrogels

    Applegate, Matthew B.; Alonzo, Carlo; Georgakoudi, Irene; Kaplan, David L.; Omenetto, Fiorenzo G.

    2016-06-01

    High resolution three-dimensional voids can be directly written into transparent silk fibroin hydrogels using ultrashort pulses of near-infrared (NIR) light. Here, we propose a simple finite-element model that can be used to predict the size and shape of individual features under various exposure conditions. We compare predicted and measured feature volumes for a wide range of parameters and use the model to determine optimum conditions for maximum material removal. The simplicity of the model implies that the mechanism of multiphoton induced void creation in silk is due to direct absorption of light energy rather than diffusion of heat or other photoproducts, and confirms that multiphoton absorption of NIR light in silk is purely a 3-photon process.

  8. Does Infrared Multiphoton Dissociation of Vinyl Chloride Yield Cold Vinylidene?

    Fernando, Ravin; Qu, Chen; Bowman, Joel M; Field, Robert W; Suits, Arthur G

    2015-07-01

    Velocity map imaging of the infrared multiphoton dissociation of vinyl chloride shows the formation of HCl in rotational levels below J = 10 that are associated with the three-center elimination pathway. The total translational energy release is observed to peak at 3-5 kcal/mol, which is consistent with the low reverse barrier predicted for the formation of HCl with vinylidene coproducts. Direct dynamics trajectory studies from the three-center transition state reproduce the observed distributions and show that the associated vinylidene is formed with only modest rotational excitation, precluding Coriolis-induced mixing among the excited vibrational levels of acetylene that would lead to distribution of vinylidene character into many vibrationally mixed acetylene vibrational levels. The results suggest that infrared multiphoton dissociation of vinyl chloride is an efficient route to synthesis of stable, cold vinylidene. PMID:26266719

  9. Individual bioaerosol particle discrimination by multi-photon excited fluorescence

    Kiselev, Denis; Bonacina, Luigi; Wolf, Jean-Pierre

    2011-01-01

    Femtosecond laser induced multi-photon excited fluorescence (MPEF) from individual airborne particles is tested for the first time for discriminating bioaerosols. The fluorescence spectra, analysed in 32 channels, exhibit a composite character originating from simultaneous two-photon and three-photon excitation at 790 nm. Simulants of bacteria aggregates (clusters of dyed polystyrene microspheres) and different pollen particles (Ragweed, Pecan, Mulberry) are clearly discriminated by their MPE...

  10. Relaxation channels of multi-photon excited xenon clusters

    Serdobintsev, P. Yu.; Melnikov, A. S. [Institute of Nanobiotechnologies, Peter the Great St.Petersburg Polytechnic University, Saint Petersburg 195251 (Russian Federation); Department of Physics, St. Petersburg State University, Saint Petersburg 198904 (Russian Federation); Rakcheeva, L. P., E-mail: lida@nanobio.spbstu.ru; Murashov, S. V.; Khodorkovskii, M. A. [Institute of Nanobiotechnologies, Peter the Great St.Petersburg Polytechnic University, Saint Petersburg 195251 (Russian Federation); Lyubchik, S. [REQUIMTE, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica 2829-516 (Portugal); Timofeev, N. A.; Pastor, A. A. [Department of Physics, St. Petersburg State University, Saint Petersburg 198904 (Russian Federation)

    2015-09-21

    The relaxation processes of the xenon clusters subjected to multi-photon excitation by laser radiation with quantum energies significantly lower than the thresholds of excitation of atoms and ionization of clusters were studied. Results obtained by means of the photoelectron spectroscopy method showed that desorption processes of excited atoms play a significant role in the decay of two-photon excited xenon clusters. A number of excited states of xenon atoms formed during this process were discovered and identified.

  11. Dark gamma-ray bursts: possible role of multiphoton processes

    Perel'man, Mark E

    2009-01-01

    The absence of optical afterglow at some gamma-ray bursts (so called dark bursts) requires analyses of physical features of this phenomenon. It is shown that such singularity can be connected with multiphoton processes of frequencies summation in the Rayleigh- Jeans part of spectra, their pumping into higher frequencies. It can be registered most probably on young objects with still thin plasma coating, without further thermalization, i.e. soon after a prompt beginning of the explosive activity.

  12. Multiphoton ionization and stabilization of helium in superintense xuv fields

    Sørngård, S. A.; Askeland, S.; Nepstad, R.; Førre, M.

    2011-01-01

    Multiphoton ionization of helium is investigated in the superintense field regime, with particular emphasis on the role of the electron-electron interaction in the ionization and stabilization dynamics. To accomplish this, we solve ab initio the time-dependent Schr\\"odinger equation with the full electron-electron interaction included. By comparing the ionization yields obtained from the full calculations with corresponding results of an independent-electron model, we come to the somewhat cou...

  13. Multiphoton ionization and stabilization of helium in superintense xuv fields

    Sørngård, Stian Astad; Askeland, Sigurd; Nepstad, Raymond; Førre, Morten

    2011-01-01

    Multiphoton ionization of helium is investigated in the superintense field regime, with particular emphasis on the role of the electron-electron interaction in the ionization and stabilization dynamics. To accomplish this, we solve ab initio the time-dependent Schr¨odinger equation with the full electron-electron interaction included. By comparing the ionization yields obtained from the full calculations with the corresponding results of an independent-electron model, we come to t...

  14. Multiphoton resonance ionization for hydrogen atom in laser field

    The Schroedinger equation of hydrogen atom in laser field is expanded by Floquet wave and can be solved by the iterative method. The atomic ionization by laser field is a complex eigenvalue problem, which is formed from differential equation and boundary condition. Then the formula of the multiphoton resonance ionization in a linear polarization laser field was obtained and it is compared with the experiment

  15. Tagging multiphoton ionization events by two-dimensional photoelectron spectroscopy

    de Groot, Mattijs; Broos, Jaap; Buma, Wybren Jan

    2007-01-01

    Two-dimensional photoelectron spectroscopy has been used to supply process-specific labels to multiphoton ionization events. Employing these tags, the authors can construct excitation and photoelectron spectra along predefined excitation routes in the neutral manifold and ionization routes to the ionic manifold from one single two-dimensional photoelectron spectrum. These results offer a novel way to elucidate the vibronic and dynamic properties of excited and ionic states. (c) 2007 American ...

  16. CHIRAL MULTIPHOTON ABSORPTION AND INVERSE SKIN EFFECT IN WLAN SYSTEMS

    Héctor Torres Silva; Mario Zamorano Lucero

    2005-01-01

    A model formed by chiral bioplasma with a set of macromolecules of DNA, which represents the human head inner structure, makes possible to analyze its behavior, when it is radiated by a microwave electromagnetic field from cellular phones and WLAN's at frequencies of 2.4 and 5.2 GHz is presented. The finite difference time domain, FDTD, numeric technique is used under multiphoton regime deduced from Maxwell equations. The numerical results of the Specific Absorption Rate, SAR, show the SAR be...

  17. Multiphoton ionization mass spectrometry of nitrated polycyclic aromatic hydrocarbons.

    Tang, Yuanyuan; Imasaka, Tomoko; Yamamoto, Shigekazu; Imasaka, Totaro

    2015-08-01

    In order to suppress the fragmentation and improve the sensitivity for determination of nitrated polycyclic aromatic hydrocarbons (NPAHs), the mechanism of multiphoton ionization was studied for the following representative NPAHs, 9-nitroanthracene, 3-nitrofluoranthene, and 1-nitropyrene. The analytes were extracted from the PM2.5 on the sampling filter ultrasonically, and were measured using gas chromatography/multiphoton ionization/time-of-flight mass spectrometry with a femtosecond tunable laser in the range from 267 to 405 nm. As a result, a molecular ion was observed as the major ion and fragmentation was suppressed at wavelengths longer than 345 nm. Furthermore, the detection limit measured at 345 nm was measured to be the subpicogram level. The organic compounds were extracted from a 2.19 mg sample of particulate matter 2.5 (PM2.5), and the extract was subjected to multiphoton ionization mass spectrometry after gas chromatograph separation. The background signals were drastically suppressed at 345 nm, and the target NPAHs, including 9-nitroanthracene and 1-nitropyrene, were detected, and their concentrations were determined to be 5 and 3 pg/m(3), respectively. PMID:26048831

  18. Multiphoton ionization studies of laser induced chemistry in clusters

    Three examples are presented where multiphoton ionization mass spectrometry is used to study photochemistry in clusters. In the first, NO+(N2O3)m and NO2+(N2O3)m are made by 226nm multiphoton ionization of the clusters produced in an expansion of NO/CH4/Ar with a trace of H2O. Second, H3O+(H2O)n and CH3OH2+(CH3OH)n are observed when sufficiently large clusters of NO(H2O)m and NO(CH3OH)m are ionized, suggesting laser initiation of intracluster charge transfer reactions in these systems. Thirdly, multiphoton ionization of mixed expansions of NO and Fe(CO)5 leads to the production of (Fe)m+, (Fe)m+(CO)n, Fe+(NO)(CO), Fe+NO, and FeO+. The mechanisms for formation of these species will be discussed and analogies drawn between intracluster and collisional chemistry. copyright 1997 American Institute of Physics

  19. Record Multiphoton Absorption Cross-Sections by Dendrimer Organometalation.

    Simpson, Peter V; Watson, Laurance A; Barlow, Adam; Wang, Genmiao; Cifuentes, Marie P; Humphrey, Mark G

    2016-02-12

    Large increases in molecular two-photon absorption, the onset of measurable molecular three-photon absorption, and record molecular four-photon absorption in organic π-delocalizable frameworks are achieved by incorporation of bis(diphosphine)ruthenium units with alkynyl linkages. The resultant ruthenium alkynyl-containing dendrimers exhibit strong multiphoton absorption activity through the biological and telecommunications windows in the near-infrared region. The ligated ruthenium units significantly enhance solubility and introduce fully reversible redox switchability to the optical properties. Increasing the ruthenium content leads to substantial increases in multiphoton absorption properties without any loss of optical transparency. This significant improvement in multiphoton absorption performance by incorporation of the organometallic units into the organic π-framework is maintained when the relevant parameters are scaled by molecular weights or number of delocalizable π-electrons. The four-photon absorption cross-section of the most metal-rich dendrimer is an order of magnitude greater than the previous record value. PMID:26797727

  20. From morphology to biochemical state – intravital multiphoton fluorescence lifetime imaging of inflamed human skin

    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.

  1. Virtual Hematoxylin and Eosin Transillumination Microscopy Using Epi-Fluorescence Imaging

    Husvogt, Lennart; Vardeh, Hilde; Faulkner-Jones, Beverly E.; Hornegger, Joachim; Connolly, James L.; Fujimoto, James G.

    2016-01-01

    We derive a physically realistic model for the generation of virtual transillumination, white light microscopy images using epi-fluorescence measurements from thick, unsectioned tissue. We demonstrate this technique by generating virtual transillumination H&E images of unsectioned human breast tissue from epi-fluorescence multiphoton microscopy data. The virtual transillumination algorithm is shown to enable improved contrast and color accuracy compared with previous color mapping methods. Finally, we present an open source implementation of the algorithm in OpenGL, enabling real-time GPU-based generation of virtual transillumination microscopy images using conventional fluorescence microscopy systems. PMID:27500636

  2. Intravital microscopy

    Masedunskas, Andrius; Milberg, Oleg; Porat-Shliom, Natalie; Sramkova, Monika; Wigand, Tim; Amornphimoltham, Panomwat; Weigert, Roberto

    2012-01-01

    Intravital microscopy is an extremely powerful tool that enables imaging several biological processes in live animals. Recently, the ability to image subcellular structures in several organs combined with the development of sophisticated genetic tools has made possible extending this approach to investigate several aspects of cell biology. Here we provide a general overview of intravital microscopy with the goal of highlighting its potential and challenges. Specifically, this review is geared...

  3. Generation of Multi-Photon Cluster States through the Cavity Input-Output Process

    We propose a scheme to generate the multi-photon cluster states via the cavity input-output process and the single-bit rotations. The method can be generalized to construct a series of multi-photon graph states, and the successful probability is close to unity in the ideal condition

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

    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. PMID:26830089

  5. Correlative microscopy.

    Loussert Fonta, Céline; Humbel, Bruno M

    2015-09-01

    In recent years correlative microscopy, combining the power and advantages of different imaging system, e.g., light, electrons, X-ray, NMR, etc., has become an important tool for biomedical research. Among all the possible combinations of techniques, light and electron microscopy, have made an especially big step forward and are being implemented in more and more research labs. Electron microscopy profits from the high spatial resolution, the direct recognition of the cellular ultrastructure and identification of the organelles. It, however, has two severe limitations: the restricted field of view and the fact that no live imaging can be done. On the other hand light microscopy has the advantage of live imaging, following a fluorescently tagged molecule in real time and at lower magnifications the large field of view facilitates the identification and location of sparse individual cells in a large context, e.g., tissue. The combination of these two imaging techniques appears to be a valuable approach to dissect biological events at a submicrometer level. Light microscopy can be used to follow a labelled protein of interest, or a visible organelle such as mitochondria, in time, then the sample is fixed and the exactly same region is investigated by electron microscopy. The time resolution is dependent on the speed of penetration and fixation when chemical fixatives are used and on the reaction time of the operator for cryo-fixation. Light microscopy can also be used to identify cells of interest, e.g., a special cell type in tissue or cells that have been modified by either transfections or RNAi, in a large population of non-modified cells. A further application is to find fluorescence labels in cells on a large section to reduce searching time in the electron microscope. Multiple fluorescence labelling of a series of sections can be correlated with the ultrastructure of the individual sections to get 3D information of the distribution of the marked proteins: array

  6. Investigation of prostate cancer cells using NADH and Tryptophan as biomarker: multiphoton FLIM-FRET microscopy

    Rehman, Shagufta; O'Melia, Meghan J.; Wallrabe, Horst; Svindrych, Zdenek; Chandra, Dhyan; Periasamy, Ammasi

    2016-03-01

    Fluorescence Lifetime Imaging (FLIM) can be used to understand the metabolic activity in cancer. Prostate cancer is one of the leading cancers in men in the USA. This research focuses on FLIM measurements of NAD(P)H and Tryptophan, used as biomarkers to understand the metabolic activity in prostate cancer cells. Two prostate cancers and one normal cell line were used for live-cell FLIM measurements on Zeiss780 2P confocal microscope with SPCM FLIM board. Glucose uptake and glycolysis proceeds about ten times faster in cancer than in non-cancerous tissues. Therefore, we assessed the glycolytic activity in the prostate cancer in comparison to the normal cells upon glucose stimulation by analyzing the NAD(P)H and Trp lifetime distribution and efficiency of energy transfer (E%). Furthermore, we treated the prostate cancer cells with 1μM Doxorubicin, a commonly used anti-cancer chemotherapeutic. Increase in NADH a2%, an indicator of increased glycolysis and increased E% between Trp and NAD(P)H were seen upon glucose stimulation for 30min. The magnitude of shift to the right for NAD(P)H a2% and E% distribution was higher in prostate cancer versus the normal cells. Upon treatment with Doxorubicin decrease in cellular metabolism was seen at 15 and 30 minutes. The histogram for NAD(P)H a2% post-treatment for prostate cancer cells showed a left shift compared to the untreated control suggesting decrease in glycolysis and metabolic activity opposite to what was observed after glucose stimulation. Hence, NAD(P)H and Trp lifetimes can be used biomarkers to understand metabolic activity in prostate cancer and upon chemotherapeutic interventions.

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

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

    2013-01-01

    -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...... excised skin, including applications of fluctuation correlation spectroscopy on transdermal penetration of liposomes are presented and discussed. The data from the different studies reported reveal the intrinsic heterogeneity of skin and also prove these strategies to be powerful noninvasive tools to...

  8. Non-perturbative methods applied to multiphoton ionization

    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)

  9. Multiphoton ionization of magnesium via an autoionizing state

    Druten, van, N.J.; Trainham, R.; Muller, H.G.

    1994-01-01

    Multiphoton single and double ionization of magnesium was studied by measuring electron energy spectra and ion mass spectra using 1-ps laser pulses in the 580-595-nm wavelength and 1012-1013-W/cm2 intensity range. In single ionization the (3p)2 1S doubly excited autoionizing state, resonant at the four-photon level, is found to play an important role. Single ionization leaving the Mg+ ion in the 3p excited state is strongly enhanced when resonant with the (3p)2 1S state. The amount of above t...

  10. Partitioning of the linear photon momentum in multiphoton ionization

    Smeenk, C; Zhou, B; Mysyrowicz, A; Villeneuve, D M; Staudte, A; Corkum, P B

    2011-01-01

    The balance of the linear photon momentum in multiphoton ionization is studied experimentally. In the experiment argon and neon atoms are singly ionized by circularly polarized laser pulses with a wavelength of 800 nm and 1400 nm in the intensity range of 10^{14} - 10^{15} W/cm^2. The photoelectrons are measured using velocity map imaging. We find that the photoelectrons carry linear momentum corresponding to the photons absorbed above the field free ionization threshold. Our finding has implications for concurrent models of the generation of terahertz radiation in filaments.

  11. Circular dichroism in XUV + IR multiphoton ionization of atoms

    Circular dichroism (CD) is theoretically considered for two-colour multiphoton ionization of arbitrary atomic subshells. In particular, p-subshell ionization is analysed and compared with s-subshell ionization. Simple analytical expressions for the CD are obtained for both s- and p-subshell ionization. The calculations performed for Ne(2p) ionization by an extreme ultraviolet pulse in the presence of an infrared laser pulse show that the CD in this case is appreciably larger than in previously discussed s-shell ionization. It makes this case favourable for applications as a sensitive tool for measuring the helicity of short-wavelength free-electron laser beams. (paper)

  12. NON-PERTURBATIVE METHODS APPLIED TO MULTIPHOTON IONIZATION

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

    1982-01-01

    We discuss the use of non-perturbative methods in the treatment of atomic ionization. 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 scheme is suggested. A brief comparison betw...

  13. Multiphoton production at high energies in the Standard Model, 1

    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 obtain recursion relations for currents consisting of a charged scalar, spinor, or vector line that radiates $n$ photons. Closed form solutions to these recursion relations for arbitrary $n$ are presented for the cases of like-helicity and one unlike-helicity photon production. We apply the currents singly and in pairs to obtain amplitudes for processes involving the production of $n$ photons with up to two unlike helicities from a pair of charged particles. The replacement of one or more photons by transversely polarized

  14. Multiphoton double ionization via field-independent resonant excitation

    The double ionization of xenon in the multiphoton regime has been studied at two wavelengths (0.77 and 0.79 μm) using an electron-ion coincidence technique and an intensity binned ion ratio method. Sharp resonant structures in the electron energy distribution correlated with the doubly charged ion, as well as a wavelength dependence of the Xe2+/Xe+ ratio provides new insights. A mechanism involving the shelving of population in Rydberg states followed by excitation of a core electron is proposed

  15. Advances in multi-photon processes and spectroscopy v.22

    Lin, S H; Fujimura, Y

    2014-01-01

    This volume presents the recent progress and perspective in multi-photon processes and spectroscopy of atoms, ions, molecules and solids. The subjects in the series cover the experimental and theoretical investigations in the interdisciplinary research fields of natural science including chemistry, physics, bioscience and material science. Contents:Theoretical Foundations for Exploring Quantum Optimal Control of Molecules (Tak-San Ho, Herschel Rabitz and Shih-I Chu)Intramolecular Nuclear Flux Densities (I Barth, C Daniel, E Gindensperger, J Manz, J F Pérez-To

  16. Signal enhancement in multiphoton imaging by the use of coated glass substrates.

    Lee, Sheng-Lin; Guo, Han-Wen; Chen, Yang-Fan; Dong, Chen-Yuan

    2015-09-01

    In nonlinear optical imaging of biological specimens, more than half of the generated luminescence signal is lost, when signal collection is performed in the epi-illuminated geometry. In this study, we enhanced the collected luminescence signal by the use of alternating multiply-coated layers of tantalum pentoxide (Ta2O5) and silicon dioxide (SiO2) on standard microscope cover glasses that has high transmission in the near-infrared wavelength region and high reflection of the visible, luminescence signal. Our coating is biocompatible, allows visual examination of the specimens and optimize collection of the luminescence signal. We demonstrated this approach on a number of specimens including sulforhodamine solution, fluorescence microspheres, and labeled 3T3 cells. In all cases, the use of coated cover glass enhanced signal, optimally by a factor of about 2. Image analysis of labeled 3T3 cells also shows signal enhancement did not contribute to additional photobleaching. Our results show that properly designed coated cover glass can enhance detected signal in multiphoton microscopy and result in improved image quality. PMID:26417521

  17. Combined multiphoton imaging and automated functional enucleation of porcine oocytes using femtosecond laser pulses

    Kuetemeyer, Kai; Lucas-Hahn, Andrea; Petersen, Bjoern; Lemme, Erika; Hassel, Petra; Niemann, Heiner; Heisterkamp, Alexander

    2010-07-01

    Since the birth of ``Dolly'' as the first mammal cloned from a differentiated cell, somatic cell cloning has been successful in several mammalian species, albeit at low success rates. The highly invasive mechanical enucleation step of a cloning protocol requires sophisticated, expensive equipment and considerable micromanipulation skill. We present a novel noninvasive method for combined oocyte imaging and automated functional enucleation using femtosecond (fs) laser pulses. After three-dimensional imaging of Hoechst-labeled porcine oocytes by multiphoton microscopy, our self-developed software automatically identified the metaphase plate. Subsequent irradiation of the metaphase chromosomes with the very same laser at higher pulse energies in the low-density-plasma regime was used for metaphase plate ablation (functional enucleation). We show that fs laser-based functional enucleation of porcine oocytes completely inhibited the parthenogenetic development without affecting the oocyte morphology. In contrast, nonirradiated oocytes were able to develop parthenogenetically to the blastocyst stage without significant differences to controls. Our results indicate that fs laser systems have great potential for oocyte imaging and functional enucleation and may improve the efficiency of somatic cell cloning.

  18. Signal enhancement in multiphoton imaging by the use of coated glass substrates

    Lee, Sheng-Lin; Guo, Han-Wen; Chen, Yang-Fan; Dong, Chen-Yuan

    2015-01-01

    In nonlinear optical imaging of biological specimens, more than half of the generated luminescence signal is lost, when signal collection is performed in the epi-illuminated geometry. In this study, we enhanced the collected luminescence signal by the use of alternating multiply-coated layers of tantalum pentoxide (Ta2O5) and silicon dioxide (SiO2) on standard microscope cover glasses that has high transmission in the near-infrared wavelength region and high reflection of the visible, luminescence signal. Our coating is biocompatible, allows visual examination of the specimens and optimize collection of the luminescence signal. We demonstrated this approach on a number of specimens including sulforhodamine solution, fluorescence microspheres, and labeled 3T3 cells. In all cases, the use of coated cover glass enhanced signal, optimally by a factor of about 2. Image analysis of labeled 3T3 cells also shows signal enhancement did not contribute to additional photobleaching. Our results show that properly designed coated cover glass can enhance detected signal in multiphoton microscopy and result in improved image quality. PMID:26417521

  19. Dynamics of rotationally-resolved multiphoton ionization processes in molecules

    This dissertation presents the results of studies of several rotationally-resolved resonance enhanced multiphoton ionization (REMPI) processes in some simple molecular systems. The objective of these studies is to quantitatively identify the underlying dynamics of this highly state-specific process which utilizes the narrow bandwidth radiation of a laser to ionize a molecule by first preparing an excited state via multiphoton absorption and subsequently ionization that state before it can decay. Coupled with high-resolution photoelectron spectroscopy, REMPI is clearly an important probe of molecular excited states and their photoionization dynamics. A key feature of these studies is that they are carried out using accurate Hartree-Fock orbitals to describe the photoelectron orbitals of the molecular ions. Studies reported here include investigations of (i) ionic rotational branching ratios and their energy dependence for REMPI via the A2Σ+(3sσ) and D2Σ+(3pσ) states of NO, (ii) the influence of angular momentum constraints on branching ratios at low photoelectron energies for REMPI via low-J levels of the resonant intermediate state, (iii) the strong dependence of photoelectron angular distributions on final ionic rotational state and on the alignment in REMPI of the A2Σ+ state of NO, (iv) vibrational state dependence of ionic rotational branching ratios arising from rapid orbital evolution in resonant states (v) the influence of rovibronic interactions on the rotational branching ratios seen in REMPI via the D2Σ+(3pσ) state of NO

  20. Dynamics of single and multiphoton ionization processes in molecules

    Single-photon and resonant multiphoton ionization studies, which can now be carried out using synchrotron radiation and lasers, respectively, are providing important dynamical information on molecular photoionization. The author studied the underlying dynamical features of these ionization processes using Hartree-Fock continuum orbitals generated using the iterative Schwinger variational method for solving the photoelecttron collisional equations. The single-photon studies examine the important role that shape and autoionizing resonances play in molecular photoionization, while the multiphoton studies investigate the ionization dynamics of exited electronic states. The subtle nature of shape resonances was demonstrated in polyatomic systems such as C2H2 and C2N2, where the possibility of multiple resonances in a single channel is observed. Molecular autoionizing resonances are known to dominate regions of the photoionization spectra. The author adapted and applied a generalization of the Fano treatment for autoionization to molecular systems. Results for H2 and C2H2 autoionizing resonances are presented and discussed

  1. Multiphoton ionization and stabilization of helium in superintense xuv fields

    Sørngård, S A; Nepstad, R; Førre, M

    2011-01-01

    Multiphoton ionization of helium is investigated in the superintense field regime, with particular emphasis on the role of the electron-electron interaction in the ionization and stabilization dynamics. To accomplish this, we solve ab initio the time-dependent Schr\\"odinger equation with the full electron-electron interaction included. By comparing the ionization yields obtained from the full calculations with corresponding results of an independent-electron model, we come to the somewhat counterintuitive conclusion that the single-particle picture breaks down at superstrong field strengths. We explain this finding from the perspective of the so-called Kramers-Henneberger frame, the reference frame of a free (classical) electron moving in the field. The breakdown is tied to the fact that shake-up and shake-off processes cannot be properly accounted for in commonly used independent-electron models. In addition, we see evidence of a change from the multiphoton to the shake-off ionization regime in the energy di...

  2. Resonant enhanced multiphoton ionization studies of atomic oxygen

    Dixit, S. N.; Levin, D.; Mckoy, V.

    1987-01-01

    In resonant enhanced multiphoton ionization (REMPI), an atom absorbs several photons making a transition to a resonant intermediate state and subsequently ionizing out of it. With currently available tunable narrow-band lasers, the extreme sensitivity of REMPI to the specific arrangement of levels can be used to selectively probe minute amounts of a single species (atom) in a host of background material. Determination of the number density of atoms from the observed REMPI signal requires a knowledge of the multiphoton ionization cross sections. The REMPI of atomic oxygen was investigated through various excitation schemes that are feasible with available light sources. Using quantum defect theory (QDT) to estimate the various atomic parameters, the REMPI dynamics in atomic oxygen were studied incorporating the effects of saturation and a.c. Stark shifts. Results are presented for REMPI probabilities for excitation through various 2p(3) (4S sup o) np(3)P and 2p(3) (4S sup o) nf(3)F levels.

  3. Multiphoton ionization and stabilization of helium in superintense xuv fields

    Multiphoton ionization of helium is investigated in the superintense field regime, with particular emphasis on the role of the electron-electron interaction in the ionization and stabilization dynamics. To accomplish this, we solve ab initio the time-dependent Schroedinger equation with the full electron-electron interaction included. By comparing the ionization yields obtained from the full calculations with the corresponding results of an independent-electron model, we come to the somewhat counterintuitive conclusion that the single-particle picture breaks down at superstrong field strengths. We explain this finding from the perspective of the so-called Kramers-Henneberger frame, the reference frame of a free (classical) electron moving in the field. The breakdown is tied to the fact that shake-up and shake-off processes cannot be properly accounted for in commonly used independent-electron models. In addition, we see evidence of a change from the multiphoton to the shake-off ionization regime in the energy distributions of the electrons. From the angular distribution, it is apparent that the correlation is an important factor even in this regime.

  4. Ultrafast multiphoton transient absorption of {beta}-carotene

    Buckup, Tiago [Philipps University Marburg, Department of Chemistry, D-35043 Marburg (Germany); Ruprecht-Karls University Heidelberg, Physical-Chemistry Institute, D-69120 (Germany); Weigel, Alexander; Hauer, Juergen [Philipps University Marburg, Department of Chemistry, D-35043 Marburg (Germany); Motzkus, Marcus, E-mail: Marcus.Motzkus@pci.uni-heidelberg.de [Philipps University Marburg, Department of Chemistry, D-35043 Marburg (Germany); Ruprecht-Karls University Heidelberg, Physical-Chemistry Institute, D-69120 (Germany)

    2010-07-19

    Multiphoton spectroscopy is able to directly excite electronic states, which are one-photon forbidden. Under single photon conditions, such one-photon forbidden states are exclusively populated via internal relaxation. Hence, transient absorption with two-photon excitation has the potential of clarifying complex relaxation networks by using aimed excitation. In this work we exploited ultrafast two-photon spectroscopy to investigate the excitation of dark states of {beta}-carotene in solution. After direct excitation of the vibronic manifold of S{sub 1}(2A{sub g}{sup -}) from S{sub 0} via two-photon transition, the characteristic internal conversion via hot-S{sub 1} {yields} S{sub 1} {yields} S{sub 0} was observed in the respective spectral region. Additional slow dynamics in the blue-wing of excited-state absorption (ESA) and in the NIR were detected, which is not directly observable with one-photon excitation transient absorption. These features are associated here to resonant multiphoton processes, which lead simultaneously to ultrafast intersystem crossing between singlet and triplet systems as well as to excitation of doublet states. Furthermore, we identify a 340-400 fs relaxation component in the near-infrared region after two-photon resonant excitation and discuss the role of additional dark states (3A{sub g}{sup -} and 1B{sub u}{sup -}) in this process.

  5. The cross-over from tunnelling to multiphoton ionization of atoms

    Klaiber, Michael

    2016-01-01

    We present a theory illuminating the cross-over from strong-field tunnelling ionization to weak-field multiphoton ionization in the interaction of a classical laser field with a hydrogen atom. A simple formula is derived in which the ionization amplitude appears as a product of two separate amplitudes. The first describes the initial polarization of the atom by virtual multiphoton absorption and the second the subsequent tunnelling out of the polarized atom. Tunnelling directly from the ground state and multiphoton absorption without tunnelling appear naturally as the limits of the theory.

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

    The favorable properties of the infrared multiphoton absorption and dissociation of trichloroethylene-H, (C2 HCl3), by TEA-CO2 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% D2 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 C2 HCl3 in isotopic mixture with C2 DCl3. (Author)

  7. Generating Nanostructures with Multiphoton Absorption Polymerization using Optical Trap Assisted Nanopatterning

    Tsai, Yu-Cheng; Leitz, Karl-Heinz; Fardel, Romain; Schmidt, Michael; Arnold, Craig B.

    The need to generate sub 100 nm features is of interest for a variety of applications including optics, optoelectronics, and plasmonics. To address this requirement, several advanced optical lithography techniques have been developed based on either multiphoton absorption polymerization or near-field effects. In this paper, we combine strengths from multiphoton absorption and near field using optical trap assisted nanopatterning (OTAN). A Gaussian beam is used to position a microsphere in a polymer precursor fluid near a substrate. An ultrafast laser is focused by that microsphere to induce multiphoton polymerization in the near field, leading additive direct-write nanoscale processing.

  8. Hamiltonian formulation for the theory of multiphoton processes in atoms based on the first principles--

    Bakasov, A.A. (Joint Institute for Nuclear Research, Head Post Office, P.O. Box 79, Moscow (SU))

    1989-07-01

    A Hamiltonian is derived on the basis of the first principles of quantum electrodynamics. The Hamiltonian is seen to describe two- and multiphoton processes. A problem of consequent derivation and microscopic substantiation of models of multiphoton processes widely used in quantum optics is solved. The first correction to the Pauli equation is obtained. The constant of interaction of a two-level atom with two photons is given in an explicit form. A method of calculating interaction constants for multilevel atoms with multiphoton transitions is presented. Other results obtained on the basis of the developed approach are discussed.

  9. Hamiltonian models of multiphoton processes and four--photon squeezed states via nonlinear canonical transformations

    De Siena, S; Illuminati, F; Siena, Silvio De; Lisi, Antonio Di; Illuminati, Fabrizio

    2002-01-01

    We introduce nonlinear canonical transformations that yield effective Hamiltonians of multiphoton down conversion processes, and we define the associated non-Gaussian multiphoton squeezed states as the coherent states of the multiphoton Hamiltonians. We study in detail the four-photon processes and the associated non-Gaussian four-photon squeezed states. The realization of squeezing, the behavior of the field statistics, and the structure of the phase space distributions show that these states realize a natural four-photon generalization of the two-photon squeezed states.

  10. Recent developments in fluorescence-based microscopy applied in biomedical sciences

    2001-01-01

    The present short review aims to give an overview of the most recent de velopments in fluorescence microscopy and its applications in biomedical science s. Apart from improvements in well-established methods based on conventional fl u orescence microscopy and confocal microscopy (fluorescence in situ hybridisa tion (FISH), tyramide signal amplification (TSA) in immunocytochemistry, new fluorop hores), more recently introduced techniques like fluorescence resonance energy t ransfer (FRET), fluorescence recovery after photobleaching (FRAP), multiphoton m icroscopy and fluorescence correlation spectroscopy (FCS) will be discussed.

  11. Evaluation of Elastin/Collagen Content in Human Dermis in-Vivo by Multiphoton Tomography—Variation with Depth and Correlation with Aging

    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.

  12. Monitoring femtosecond laser microscopic photothermolysis with multimodal microscopy (Conference Presentation)

    Huang, Yimei; Lui, Harvey; Zhao, Jianhua; McLean, David I.; Zeng, Haishan

    2016-02-01

    Photothermolysis induced by femtosecond (fs) lasers may be a promising modality in dermatology because of its advantages of high precision due to multiphoton absorption and deeper penetration due to the use of near infrared wavelengths. Although multiphoton absorption nonlinear effects are capable of precision targeting, the femtosecond laser photothermolysis could still have effects beyond the targeted area if a sufficiently high dose of laser light is used. Such unintended effects could be minimized by real time monitoring photothermolysis during the treatment. Targeted photothermolytic treatment of ex vivo mouse skin dermis was performed with tightly focused fs laser beams. Images of reflectance confocal microscopy (RCM), second harmonic generation (SHG), and two-photon fluorescence (TPF) of the mouse skins were obtained with integrated multimodal microscopy before, during, and after the laser treatment. The RCM, SHG, and TPF signal intensities of the treatment areas changed after high power femtosecond laser irradiation. The intensities of the RCM and SHG signals decreased when the tissue was damaged, while the intensity of the TPF signal increased when the photothermolysis was achieved. Moreover, the TPF signal was more susceptible to the degree of the photothermolysis than the RCM and SHG signals. The results suggested that multimodal microscopy is a potentially useful tool to monitor and assess the femtosecond laser treatment of the skin to achieve microscopic photothermolysis with high precision.

  13. Two-photon excitation improves multifocal structured illumination microscopy in thick scattering tissue

    Ingaramo, Maria; York, Andrew G.; Wawrzusin, Peter; Milberg, Oleg; Hong, Amy; Weigert, Roberto; Shroff, Hari; Patterson, George H.

    2014-01-01

    Superresolution microscopy has made much progress in improving resolution and imaging speed over the past several years, but the ability to image below the diffraction limit in thick scattering specimens has not kept pace. In many interesting samples, such as Caenorhabditis elegans, Drosophila melanogaster, mouse, or human tissues, resolution is limited primarily by scattering rather than diffraction. In this paper, we show that the combination of multiphoton excitation with multifocal struct...

  14. Multiphoton ionization of CF3I clusters by ultraviolet laser radiation

    The results of the investigation of the multiphoton ionization of (CF3I)n clusters by ultraviolet laser radiation are reported. The yields of the I2+ and I+ ions, which are the products of the multiphoton ionization, have been measured as functions of the intensity of the ultraviolet radiation at the wavelengths of 308 and 232.5 nm. The degree of multiphoton ionization has been determined and appears to depend on the wavelength of radiation. The velocity distributions of the products have been measured in detail for various wavelengths and various polarizations of radiation. The anisotropy parameters of the velocity distributions of the produced ions and their kinetic energy have been determined. After analysis of the data, a mechanism of the multiphoton ultraviolet ionization of the clusters under investigation has been proposed. This mechanism depends on the used wavelengths.

  15. Intensity and Polarization Effects in Short-Pulse Multiphoton Ionization of Xenon

    KANG Hui-Peng; WANG Chuan-Liang; LIN Zhi-Yang; CHEN Yong-Ju; WU Ming-Yan; QUAN Wei; LIU Hong-Ping; LIU Xiao-Jun

    2011-01-01

    @@ We present photoelectron spectra (PES) of xenon subject to ultrashort intense laser pulses at 400nm.The intensity-dependent PES exhibit the dominance of ac-Stark-shifted multiphoton resonances in a multiphoton ionization process.A distinct difference in the spectra with different laser polarization states (i.e., linearly and circularly polarized states) is revealed and can be understood in terms of the quantum selection rule, whichrestricts the angular momentum of states that may shift into multiphoton resonances.Furthermore, the intensity dependence of the resonance-enhanced electron yield is analyzed in the context of multiphoton Landau-Zener theory.The model calculation results considering the focal volume effect are in good agreement with the experimental observation.

  16. Intensity and Polarization Effects in Short-Pulse Multiphoton Ionization of Xenon

    We present photoelectron spectra (PES) of xenon subject to ultrashort intense laser pulses at 400 nm. The intensity-dependent PES exhibit the dominance of ac-Stark-shifted multiphoton resonances in a multiphoton ionization process. A distinct difference in the spectra with different laser polarization states (i.e., linearly and circularly polarized states) is revealed and can be understood in terms of the quantum selection rule, which restricts the angular momentum of states that may shift into multiphoton resonances. Furthermore, the intensity dependence of the resonance-enhanced electron yield is analyzed in the context of multiphoton Landau-Zener theory. The model calculation results considering the focal volume effect are in good agreement with the experimental observation. (atomic and molecular physics)

  17. Wavelength scaling of high-harmonic generation efficiency close to the multiphoton ionization regime

    Lai, Chien-Jen; Cirmi, Giovanni; Huang, Shu-Wei; Granados, Eduardo; Hong, Kyung-Han; Moses, Jeffrey; Keathley, Philip; Bhardwaj, Siddharth; Kaertner, Franz

    2013-01-01

    Our experiment shows a less dramatic wavelength scaling of high harmonic generation efficiency between the tunneling and the multiphoton ionization regimes, which can be explained by a modified three-step model with complex ionization time.

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

    Dell'Anno, F; Illuminati, F; Anno, Fabio Dell'; Siena, Silvio De; 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 mixings 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 to vary at will the statist...

  19. Properties of Differential Scattering Section Based on Multi-photon Nonlinear Compton Effect

    2002-01-01

    Properties of damping electrons in collision with photons based on multi-photon nonlinear Compton effect are investigated. The expressions of the differential scattering section are derived. Several useful conclusions are drawn.

  20. Real-time high dynamic range laser scanning microscopy

    Vinegoni, C.; Leon Swisher, C.; Fumene Feruglio, P.; Giedt, R. J.; Rousso, D. L.; Stapleton, S.; Weissleder, R.

    2016-04-01

    In conventional confocal/multiphoton fluorescence microscopy, images are typically acquired under ideal settings and after extensive optimization of parameters for a given structure or feature, often resulting in information loss from other image attributes. To overcome the problem of selective data display, we developed a new method that extends the imaging dynamic range in optical microscopy and improves the signal-to-noise ratio. Here we demonstrate how real-time and sequential high dynamic range microscopy facilitates automated three-dimensional neural segmentation. We address reconstruction and segmentation performance on samples with different size, anatomy and complexity. Finally, in vivo real-time high dynamic range imaging is also demonstrated, making the technique particularly relevant for longitudinal imaging in the presence of physiological motion and/or for quantification of in vivo fast tracer kinetics during functional imaging.

  1. Dynamics of multi-photon photoluminescence in gold nanoantennas

    Biagioni, P; Huang, J -S; Kern, J; Duò, L; Hecht, B; Finazzi, M; Cerullo, G

    2011-01-01

    We perform a combined study of the degree of nonlinearity and the temporal dynamics of multiphoton-excited photoluminescence (MPPL) in gold nanoantennas. At variance with standard gold two-photon photoluminescence (TPPL), the large photoluminescence enhancement in resonant nanostructures is sometimes found to involve more than two absorbed photons per emitted photon. By two-pulse correlation measurements of TPPL and MPPL we are able to address the particular dynamics of these mechanisms. Our data give direct evidence for the two-step model that has been proposed for gold TPPL and suggest that MPPL is characterized by very similar dynamics. Interestingly, for resonant antennas we observe a reduced MPPL relaxation time compared to off-resonant antennas.

  2. Clinical multiphoton tomography and clinical two-photon microendoscopy

    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.

  3. Tunneling dynamics in multiphoton ionization and attoclock calibration

    Klaiber, M; Keitel, C H

    2014-01-01

    The intermediate domain of strong-field ionization between the tunneling and the multiphoton regimes is investigated using the strong field approximation and the imaginary-time method. An intuitive model for the dynamics is developed which describes the ionization process within a nonadiabatic tunneling picture with a coordinate dependent electron energy during the under-the-barrier motion. The nonadiabatic effects in the elliptically polarized laser field induce a transversal momentum shift of the tunneled electron wave packet at the tunnel exit, a delayed appearance in the continuum as well as a shift of the tunneling exit towards the ionic core. The latter significantly modifies the Coulomb focusing during the electron excursion in the laser field after exiting the ionization tunnel. We show that nonadiabatic effects are especially large when the Coulomb field of the ionic core is taken into account during the under-the-barrier motion. The simpleman model modified with these nonadiabatic corrections provid...

  4. Multiphoton ionization of acetone-water clusters at 355 nm

    WANG Reng; KONG Xiang-he; ZHANG Shu-dong; ZHANG Xia; FAN Xing-yan; ZHAO Shu-yan

    2006-01-01

    @@ The multiphoton ionization of acetone-water clusters were detected at 355 nm laser wavelength by using the time of flight mass spectrometer(TOF-MS).The experiments show that all products are protonated.Three main products such as (CH3COCH3)n-(H2O)n-2H+,(CH3COCH3)n-(H2O)n-1H+ and (CH3COCH3)n-(H2O)nH+ are concluded from the results.In order to study the equilibrium structures of the (CH3COCH3)n-(H2O)n-2H+,the ab-initio calculation is used on them.The experiment is even done when the volume rate of acetone to water is 1:2.

  5. Multiphoton production at high energies in the Standard Model, 2

    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, longitudinal $Z$ or neutrino pairs.

  6. Quadrature-dependent Bogoliubov transformations and multiphoton squeezed states

    De Siena, S; Illuminati, F; Siena, Silvio De; Lisi, Antonio Di; Illuminati, Fabrizio

    2001-01-01

    We introduce a linear, canonical transformation of the fundamental single--mode field operators $a$ and $a^{\\dagger}$ that generalizes the linear Bogoliubov transformation familiar in the construction of the harmonic oscillator squeezed states. This generalization is obtained by adding to the linear transformation a nonlinear function of any of the fundamental quadrature operators $X_{1}$ and $X_{2}$, making the original Bogoliubov transformation quadrature--dependent. Remarkably, the conditions of canonicity do not impose any constraint on the form of the nonlinear function, and lead to a set of nontrivial algebraic relations between the $c$--number coefficients of the transformation. We examine in detail the structure and the properties of the new quantum states defined as eigenvectors of the transformed annihilation operator $b$. These eigenvectors define a class of multiphoton squeezed states. The structure of the uncertainty products and of the quasiprobability distributions in phase space shows that bes...

  7. Monitoring wound healing by multiphoton tomography/endoscopy

    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.

  8. Picosecond multiphoton ionization of atomic and molecular clusters

    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

  9. Partial indistinguishability theory for multi-photon experiments in multiport devices

    Shchesnovich, V. S.

    2014-01-01

    We generalize an approach for description of multi-photon experiments with multi-port unitary linear optical devices, initiated in \\textit{Phys. Rev. A \\textbf{89}, 022333 (2014)} for the case of single photons in mixed spectral states, to arbitrary (multi-photon) input and arbitrary photon detectors. We give a physical interpretation of a non-negative definite Hermitian matrix, the matrix of a quadratic form giving output probabilities, as the partial indistinguishability matrix. We show tha...

  10. Multiphoton Rabi Oscillations of Correlated Electrons in Strong Field Nonsequential Double Ionization

    Qing, Liao; Yueming, Zhou; Cheng, Huang; Peixiang, Lu

    2011-01-01

    With quantum calculations, we have investigated the multiphoton nonsequential double ionization of helium atoms in intense laser fields at ultraviolet wavelengths. Very surprisingly, we find a so-far unobserved double-circle structure in the correlated electron momentum spectra. The double-circle structure essentially reveals multiphoton Rabi oscillations of two electrons, which are strongly supported by the oscillating population of a certain doubly excited state and by the oscillating doubl...

  11. Multiphoton atom ionization in a field of an ultrashort laser pulse

    Closed analytical expressions for the probability of the multiphoton atoms and ions ionization under effect of the alternating current filed applicable for any value of Keldysh parameter are obtained through imaginary time method. Dependence of the ionization probability and photoelectron pulse spectrum on the ultrashort laser pulse form is considered. The formulae obtained may be used in the theory of the semiconductor multiphoton ionization by the laser pulse field

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

    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 SF6 caused by vibrational self-quenching. Between 1000-3000 cm-1 of energy is removed from SF6 excited to approx. > 60 kcal/mole by collision with a cold SF6 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 SiF4 as absorbing gas for the CO2 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

  13. High-order multiphoton ionization of the noble gases

    Ionization of an atom by the absorption of several photons from a strong electromagnetic field is considered. Specifically, the absolute yield of multiply charged ions of the noble gases, argon, krypton, and xenon produced using a well-characterized, tunable picosecond dye laser is reported as a function of both laser intensity and frequency. Theoretical models are developed to provide a quantitative interpretation of the data. The experiments were designed to investigate the influence of intermediate atomic states on the multiphoton ionization probability at intensities such that the strength of the applied field approached that of the internal atomic field. This was accomplished by measuring the ion yield over a broad range of laser intensity, 1012 ≤ I ≤ 4 x 1014 Wcm2 and frequency. The laser wavelength was varied between 570 and 620 nm and the second harmonic, 285 to 310 nm. Enhancement of the yield of singly-charged krypton ions by two orders of magnitude by three-photon resonant, four-photon ionization was observed at intensities nearly two orders of magnitude above previous results. A model which considers only two atomic levels coupled by the strong field is presented and is found to be in good agreement with the experimental results. Measured values of the three-photon Rabi rate to, and the photoionization cross sections of, several excited states of Kr I are reported. In addition, even at intensities exceeding 1013 Wcm2, it was possible to find frequencies at which no resonant enhancement of the ionization probability could be observed. Charge states as high as Ar4+, Kr5+ and Xe6+ produced by this non-resonant multiphoton ionization were observed at the highest intensities. 102 refs., 50 figs., 5 tabs

  14. Multiphoton photochemical crosslinking-based fabrication of protein micropatterns with controllable mechanical properties for single cell traction force measurements

    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.

  15. Discharge and Multiphoton Ionization in the Focus of Powerful Lasers

    The paper gives a review of the most interesting physical results obtained in recent years during studies of the development of light sparks and multiphoton ionization of atoms. The processes taking place in light sparks - the avalanche ionization of gases in a laser focus - are considered. The basic features of light sparks, determined by the focal length of the lens and the power of the laser beam, are indicated. All sparks can be classed as ''short'' or ''long''. In ''short'' sparks the discharge plasma can extend across the laser beam, and a large part of the light energy is absorbed in it. In ''long'' sparks, produced with lenses of long focal length and high power lasers, only a small fraction of the energy goes into the plasma. ''Short'' sparks propagate in the direction of the laser, while ''long'' sparks propagate along a straight line in two opposite directions. The sparks are characterized by a striated structure. This structure is apparently due not only to the characteristics of the plasma produced, but also to the space-time structure of the laser radiation.. Thus, the production of a light spark is much more complicated than was assumed in earlier theoretical works. In ''short'' laser sparks produced with a lens of short focal length, high temperatures are reached (approx. 106 degrees), so that they can be regarded as high-temperature explosions. To understand the processes taking place in the spark, it is important to consider the following stages:' 'the formation of a fast photo-ionization corona, the production of a shock-wave fireball, the separation of,the shock wave from the fireball, the production of two-layer structure in the shock wave and, finally, the formation of long- lived decaying plasma. At lower pressures there is no avalanche ionization, and ionization is due mainly to the multiphoton photo-effect. The first results on multiphoton ionization of atoms by a strong changing field of laser radiation were presented at the Yugoslav

  16. CHIRAL MULTIPHOTON ABSORPTION AND INVERSE SKIN EFFECT IN WLAN SYSTEMS

    Héctor Torres Silva

    2005-12-01

    Full Text Available A model formed by chiral bioplasma with a set of macromolecules of DNA, which represents the human head inner structure, makes possible to analyze its behavior, when it is radiated by a microwave electromagnetic field from cellular phones and WLAN's at frequencies of 2.4 and 5.2 GHz is presented. The finite difference time domain, FDTD, numeric technique is used under multiphoton regime deduced from Maxwell equations. The numerical results of the Specific Absorption Rate, SAR, show the SAR behavior in function of input power and the chirality factor. The main conclusions of our work are: a the microwave absorption from cellular phones or WLAN's is enhanced, compared with classical models, when values of the normalized chiral factor are of order of one which appear under multiphoton regime ; b a phenomena like an “inverse skin effect” in 5.2 GHz, with respect to a 2.4 GHz source, was observed. c In the metamaterial region we show that the absorption rate always is positive.Un modelo formado por bioplasma quiral con un conjunto de macromoléculas de ADN, que representa la estructura interna de la cabeza humana, hace posible analizar su comportamiento, cuando es irradiada por campos electromagnéticos de microondas de teléfonos celulares o sistemas WLAN a frecuencias de 2.4 y 5.2 GHz. El método de diferencias finitas en el dominio del tiempo, FDTD, en régimen de multifotones deducido de las ecuaciones de Maxwell es usado. Los resultados numéricos de la taza de absorción específica SAR, muestran el comportamiento de la SAR en función de la potencia de entrada y del factor quiral. Las principales conclusiones de nuestro trabajo son: a la absorción de microondas es aumentada comparada con modelos clásicos, cuando valores del factor quiral normalizado son del orden de la unidad, que aparecen bajo régimen multifotónico; b Un fenómeno de efecto pelicular inverso en 5.2 GHz con respecto a una fuente de 2.4 GHz fue observado; c En la regi

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

    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

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

  19. Photoionization of excited molecular states using multiphoton excitation techniques

    Photoelectron spectra are reported for three photon resonant, four photon ionization of H2 via the B 1Σ/sub u/+, v = 7 (J = 2,4) and C 1π/sub u'/, v = 0-4 (J = 1) levels and of N2 via the o31π/sub u'/, v = 1,2, b 1π/sub u'/, v = 3-5, and c 1π/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

  20. Photoionization of excited molecular states using multiphoton excitation techniques

    Dehmer, Patricia M. [Argonne National Laboratory (ANL), Argonne, IL (United States); Pratt, Stephen T. [Argonne National Laboratory (ANL), Argonne, IL (United States); Dehmer, Joseph L. [Argonne National Laboratory (ANL), Argonne, IL (United States)

    1984-01-01

    Photoelectron spectra are reported for three photon resonant, four photon ionization of H₂ via the B ¹Σu⁺, v = 7 (J = 2,4) and C ¹Πu v = 0-4 (J = 1) levels and of N₂ via the o₃ ¹Πu, v = 1,2, b ¹Πu, v = 3-5, and c ¹Πu, 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.

  1. Multi-photon Resonance Phenomena Using Laguerre-Gaussian Beams

    Kazemi, Seyedeh Hamideh

    2016-01-01

    We study the influence of Laguerre-Gaussian (LG) and Gaussian fields on the linewidth of the optical spectrum of multi-photon resonance phenomena. First, we investigate the dependence of the steady-state coherence on the laser profile in a two-level system. Thanks to the LG field, the linewidth of the one-photon optical pumping peak is explicitly narrower than for a Gaussian field. We then investigate the atomic coherence in a two-level pump-probe atomic system and show that using the LG fields, a narrower two-photon absorption peak can be obtained compared to the usual Gaussian ones. In next section, we investigate the effect of the laser profiles on the coherent population trapping in the $\\Lambda$-type molecular open systems. It is shown that, comparing with the the Gaussian fields, the LG fields reduce the linewidth of the optical spectrum. In addition, for a laser-driven four-level atomic system we study the effect of laser profiles on the Autler-Townes doublet structure in the absorption spectrum. We al...

  2. Security of quantum key distribution with multiphoton components

    Yin, Hua-Lei; Fu, Yao; Mao, Yingqiu; Chen, Zeng-Bing

    2016-07-01

    Most qubit-based quantum key distribution (QKD) protocols extract the secure key merely from single-photon component of the attenuated lasers. However, with the Scarani-Acin-Ribordy-Gisin 2004 (SARG04) QKD protocol, the unconditionally secure key can be extracted from the two-photon component by modifying the classical post-processing procedure in the BB84 protocol. Employing the merits of SARG04 QKD protocol and six-state preparation, one can extract secure key from the components of single photon up to four photons. In this paper, we provide the exact relations between the secure key rate and the bit error rate in a six-state SARG04 protocol with single-photon, two-photon, three-photon, and four-photon sources. By restricting the mutual information between the phase error and bit error, we obtain a higher secure bit error rate threshold of the multiphoton components than previous works. Besides, we compare the performances of the six-state SARG04 with other prepare-and-measure QKD protocols using decoy states.

  3. Quantum Decoherence for Multi-Photon Entangled States

    SUN Yan-Hua; ZHU Xia; KUANG Le-Man

    2005-01-01

    @@ We investigate quantum decoherence of the multi-photon entangled state |ψNm> = Nm[cosγ|N- m>1|m>2 +eiθm sinγ|m>1|N - m>2]. When the entangled channel |ψNm> is embedded in an environment, the channel decoheres and becomes a mixed state governed by a master equation. We calculate thelinear entropy and the relative entropy of entanglement, which describe the mixedness and the amount of entanglement for the mixed state, respectively. We show that quantum decoherence weakens the amount of entanglement and enhances the mixedness with the time evolution. It is indicated that the relative entropy of entanglement depends on not only the initial entanglement angle and the decohering parameter, but also the number of photons in each mode. In particular, we find that the decohering speed depends on the number-difference of photons in the two modes. The larger the number-difference of photons is, the higher the decohering speed.

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

    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...

  5. Multiphoton ionization of pyrrole-water mixed clusters

    2001-01-01

    Multiphoton ionization of the hydrogen-bonded pyrrole-water clusters ( C4H5 N)n H2O)m is studied with a reflectron—time of flight mass spectrometer at 355 nm. With increasing partial concentration of pyrrole in a gas mixture source, a series of poly-pyrrole-water binary-mixed cluster ions can be observed, including unprotonated cluster ions [(C4H5N)x (H2O)y]* , protonated cluster ions [(C4H5N)x (H2O)yH]* and dehydrogenated cluster ions [ ( C4 H4 N) ( C4 H5 N) x ( H2O) y ] + . Ab initio calculations of their structures, bond strengths, charge distributions and re action energies are carried out. Stable structures of these clusters are obtained from the calculations. A probable forma tion mechanism of the cluster ions [(C4H5N)x(H2O)y] + , [(C4H5N)x (H2O)y]H+ and [(C4H4N) (C4H5N)x (H2O)y]-is supposed to be the ionization of clusters followed by dissociation.

  6. Materials characterization using ion bombardment and multiphoton resonance ionization

    The combination of energetic ion bombardment with multiphoton resonance ionization (MPRI) spectroscopy has proven to be an important advancement in surface science. The goal of this project is continuing the development of MPRI of desorbed neutrals as a surface analytical tool. The method for accomplishing this is a detailed examination of the factors which govern a measurement and the implementation of the optimum experimental approach. Initially, a review of the progress in laser post-ionization of desorbed neutral particles is presented. This is followed by a description of the newly redesigned instrument, emphasizing detailed characterization of the high current ion source and the reflecting time-of-flight mass spectrometer. Using the new apparatus, the quantitative aspects of the measurement are examined and the fractions of desorbed ions and neutrals are determined using several matrices. The In concentration in a set of silicon wafers is measured, yielding a detection limit of 9 parts-per-trillion. Finally, the prospects for employing this experiment for measuring the half-life of the rare double beta decay of 136Xe to 136Ba are assessed

  7. Tunneling dynamics in multiphoton ionization and attoclock calibration.

    Klaiber, Michael; Hatsagortsyan, Karen Z; Keitel, Christoph H

    2015-02-27

    The intermediate domain of strong-field ionization between the tunneling and multiphoton regimes is investigated using the strong-field approximation and the imaginary-time method. An intuitive model for the dynamics is developed which describes the ionization process within a nonadiabatic tunneling picture with a coordinate dependent electron energy during the under-the-barrier motion. The nonadiabatic effects in the elliptically polarized laser field induce a transversal momentum shift of the tunneled electron wave packet at the tunnel exit and a delayed appearance in the continuum as well as a shift of the tunneling exit towards the ionic core. The latter significantly modifies the Coulomb focusing during the electron excursion in the laser field after exiting the ionization tunnel. We show that nonadiabatic effects are especially large when the Coulomb field of the ionic core is taken into account during the under-the-barrier motion. The simple man model modified with these nonadiabatic corrections provides an intuitive background for exact theories and has direct implications for the calibration of the attoclock technique. PMID:25768761

  8. Development of resonance-enhanced multiphoton ionization system

    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 cm3. The integrity of the set-up is obtained from the photodissociation dynamics studies of bromoform. (author)

  9. The polarization effect of a laser in multiphoton Compton scattering

    Liang, Guo-Hua; Lü, Qing-Zheng; Teng, Ai-Ping; Li, Ying-Jun

    2014-05-01

    The multiphoton Compton scattering in a high-intensity laser beam is studied by using the laser-dressed quantum electrodynamics (QED) method, which is a non-perturbative theory for the interaction between a plane electromagnetic field and a charged particle. In order to analyze in the real experimental condition, a Lorentz transformation for the cross section of this process is derived between the laboratory frame and the initial rest frame of electrons. The energy of the scattered photon is analyzed, as well as the cross sections for different laser intensities and polarizations and different electron velocities. The angular distribution of the emitted photon is investigated in a special velocity of the electron, in which for a fixed number of absorbed photons, the electron energy will not change after the scattering in the lab frame. We obtain the conclusion that higher laser intensities suppress few-laser-photon absorption and enhance more-laser-photon absorption. A comparison between different polarizations is also made, and we find that the linearly polarized laser is more suitable to generate nonlinear Compton scattering.

  10. Fundamentals of fluorescence microscopy exploring life with light

    Mondal, Partha Pratim

    2014-01-01

    This book starts at an introductory level and leads reader to the most advanced developments in fluorescence imaging and super-resolution techniques that have enabled the emergence of new disciplines such as nanobioimaging, multiphoton microscopy, photodynamic therapy, nanometrology and nanosensors. The interdisciplinary subject of fluorescence microscopy and imaging requires complete knowledge of imaging optics and molecular physics. So, this book approaches the subject by introducing optical imaging concepts before going deep into the advanced imaging systems and their applications. Molecular orbital theory forms the basis for understanding fluorescent molecules and thereby facilitates complete explanation of light-matter interaction at the geometrical focus. The two disciplines have some overlap since light controls the states of molecules and conversely, molecular states control the emitted light. These two mechanisms together determine essential fluorescence  factors and phenomena such as, molecular cro...

  11. In Vivo Two-Photon Microscopy of Single Nerve Endings in Skin

    Yuryev, Mikhail; Molotkov, Dmitry

    2014-01-01

    Nerve endings in skin are involved in physiological processes such as sensing1 as well as in pathological processes such as neuropathic pain2. Their close-to-surface positioning facilitates microscopic imaging of skin nerve endings in living intact animal. Using multiphoton microscopy, it is possible to obtain fine images overcoming the problem of strong light scattering of the skin tissue. Reporter transgenic mice that express EYFP under the control of Thy-1 promoter in neurons (including periphery sensory neurons) are well suited for the longitudinal studies of individual nerve endings over extended periods of time up to several months or even life-long. Furthermore, using the same femtosecond laser as for the imaging, it is possible to produce highly selective lesions of nerve fibers for the studies of the nerve fiber restructuring. Here, we present a simple and reliable protocol for longitudinal multiphoton in vivo imaging and laser-based microsurgery on mouse skin nerve endings. PMID:25178088

  12. Theory of Multiphoton Multielectron Ionization of Xenon under Strong 93-eV Radiation

    We present a theoretical interpretation of recent experimental results on multiphoton multiple ionization of xenon by soft-x-ray radiation of photon energy ∼93 eV and intensity up to 1016 W/cm2[A. A Sorokin et al., Phys. Rev. Lett. 99, 213002 (2007)]. The data are interpreted within multiphoton perturbation theory, taking into account the spatiotemporal distribution of the radiation. Multiphoton cross sections have been obtained through a technique of scaling, with occasional adjustment to the data, provided the two prove to be compatible. Whatever discrepancies between theory and experiment persist can be reasonably attributed to some uncertainty in the experimental conditions and possibly to the value of some cross sections, without, however, any evidence for nonperturbative behavior

  13. Prolate spheroidal coordinates for calculations of rates for multiphoton ionization of H2+

    The validity of using the spheroidal coordinates in the problem of multiphoton ionization of H2+ is investigated in conjunction with a method that is highly efficient in the calculation of rates for multiphoton ionization of H atom in the nonperturbative intensities of laser by resuming the Rayleigh-Schroedinger perturbation series. This method takes advantages of the separability of the Hamiltonian, which in turn enables us to calculate rates of multiphoton ionization for more complex atoms or molecules. We utilize the efficiency to apply this method to the case of H2+ and compare the results to those of H atom. This method turns out to be as efficient as in the H atom calculation over a finite range of frequencies in contrast with the failure for very high or low frequencies and the reason of which are also discussed.

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

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

    2004-01-01

    Extending the scheme developed for a single mode of the electromagnetic field in the preceding paper ``Structure of multiphoton quantum optics. I. Canonical formalism and homodyne squeezed states'', 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 non degenerate and degenerate multiphoton processes. We determine the coherent states associated to the canonical transformations, which generalize the non degenerate two--photon squeezed states. Such heterodyne multiphoton squeezed 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 non classical, entangled states. For a quadratic nonline...

  15. The effect laser pulse width on the multi-photon ionization efficiency

    We studied the variation of the multi-photon ionization efficiencies of atoms as a function of the laser pulse width under the condition of constant pulse energy. The density-matrix calculation showed that long pulses were preferred for efficient ionization whenever the lifetimes of the levels involved in the transition were longer than the pulse widths. Also, it was demonstrated that the atomic and the laser parameters, such as transition cross-sections, laser pulse widths, and lifetimes affect the optimal excitation rate of the transition for efficient multi-photon ionization

  16. Multiphoton atom ionization on the field of an ultrashort laser pulse

    One derived closed analytical expressions for probability of multiphoton ionization of atoms and ions under the effect of alternating electrical field applicable for arbitrary values of Keldysh parameter. One studies dependences of ionization probability and pulsed spectrum of photoelectrons on the shape of ultrashort laser pulse. One studied in detail examples of various type pulsed fields. One studied interference effect in energy spectrum of photoelectrons at atom ionization by general periodical field. One discusses field of application for adiabatic approximation in theory of multiphoton ionization

  17. Integral-equation approach to multiphoton ionization by intense fields. II. Application to H and H-

    We have developed an integral-equation method for treating multiphoton ionization of atomic systems irradiated by intense fields. We have applied this method to the calculation of partial rates for multiphoton detachment of H- (into specific continuum channels) by a laser operating at frequencies of 0.2 and 0.5 eV. Nonperturbative features are seen at intensities above about 5 x 10/sup 10/ W/cm2. We have also applied the method to hydrogen, to the determination of the field-induced width in the ground-state energy (the total ionization rate)

  18. Coherent Phase Control of Multiphoton Ionization in Three-Level Ladder-Type System

    ZHANG Shi-An; CHEN Yu-Ting; WANG Zu-Geng; SUN Zhen-Rong

    2009-01-01

    We present the theoretical investigation of photoelectron spectroscopy resulting from the strong field induced multiphoton ionization in a typical three-level ladder-style system.Our theoretical results show that the photoelectron spectral structure can be alternatively steered by spectral phase modulation.This physical mechanism for strong field quantum control is explicitly exploited by the time-dependent dressed state population.It is concluded that the phase-shaped laser pulses can be used to selectively manipulate the multiphoton ionization process in complicated quantum systems.

  19. Fermi-coupled spherically adapted effective states in the collisionless multiphoton excitation of SF6

    A calculation method for the collisionless multiphoton excitation of SF6 by intense CO2 laser light up to a chain of parallel nν3, (n - 1)ν3 + ν2 + ν6 ... vibrational-rotational ladders linked by Fermi interaction is described. Spherically adapted effective states suitable to the purpose are defined, and matrix elements for multiphoton excitation in the rotating wave approximation effective hamiltonian formalism are given in this basis. The method is aimed at the investigation of population transfer between the cited parallel vibrational ladders, and is suitable for computer-calculation programmation. (orig.)

  20. Fermi-coupled spherically adapted effective states in the collisionless multiphoton excitation of SF 6

    Di Lauro, C.; Lattanzi, F.

    1982-10-01

    A calculation method for the collisionless multiphoton excitation of SF 6 by intense CO 2 laser light up to a chain of parallel nv3, ( n - 1) v3 + v2 + v6 … vibrational-rotational ladders linked by Fermi interaction is described. Spherically adapted effective states suitable to the purpose are defined, and matrix elements for multiphoton excitation in the rotatingwave approximation effective hamiltonian formalism are given in this basis. The method is aimed at the investigation of population transfer between the cited parallel vibrational ladders, and is suitable for computer-calculation programmation.

  1. Infrared multiphoton dissociation processes of some halogenated hydrocarbons

    We review our studies on mechanisms and dynamics of infrared multiphoton dissociation of CF3CHClF, C2HCl3, CBrF2CHClF, CBrF2CHBrF, CBrF2CBrClF, and c-C4F8 using a photofragmentation translational spectroscopy and ab initio MO calculations. In the case of CF3CHClF, three-centered HCl elimination and C-Cl bond rupture occurred competitively as primary dissociation processes. The center-of-mass translational energy distribution of the products produced by the HCl elimination indicates that an exit barrier of several kcal/mol exists in the potential energy surface of the reaction. In the case of C2HCl3, three- and four-centered eliminations of HCl occurred competitively. Halogenated ethanes containing bromine atoms dissociated through a C-Br bond rupture channel. The halogenated ethyl radicals produced by the C-Br bond ruptures subsequently dissociated through a C-halogen bond rupture channel. The translational energy distributions for the C-halogen bond ruptures observed are almost consistent with those calculated by RRKM theory. In the case of c-C4F8, dissociation of c-C4F8 to two C2F4 molecules was observed as a primary process. About 30% of the exit barrier for this reaction is converted to the translational energy of the products. This indicates that the dissociation proceeds through a loose transition state. This conclusion is supported by the transition state structure calculated by an ab initio MO method. (author)

  2. High-intensity multiphoton ionization of H2

    A tunable, high-intensity picosecond-dye-laser system has been employed with electron energy analysis to investigate the dynamics of (3+1) resonance-enhanced multiphoton ionization via various vibrational levels of the B 1Σu+ and C 1Πu electronic states in H2. At the intensities studied [(0.2--6)x1013 W/cm2], we find evidence for production of molecular ions in various vibrational levels; at the lower intensities the population distribution of final vibrational states varies with wavelength in a manner consistent with resonant enhancement at the three-photon level, followed by ionization into a vibrational level of H2+ roughly predictable by a Franck-Condon analysis of ionization out of the C state. At higher intensities, there is a shift to increased population of lower vibrational states of H2+, consistent with an ac Stark shift of the correspondingly lower vibrational levels of the C state into resonance with the three-photon energy of the laser. Clear evidence of direct dissociation of H2 followed by single-photon ionization of the excited H atom is observed as well. Above-threshold ionization of these two processes occurs readily. We also find that dissociative ionization is an increasingly important ionization pathway as the wavelength is increased. Finally, we see evidence of a fourth ionization pathway, which we tentatively assign to photoionization into a transient bound state created by the avoided crossing of the first repulsive electronic state of H2+, |2pσu,n right-angle, with the single-photon-dressed ground state of H2+, |1sσg,n+1 right-angle

  3. Intravital multiphoton imaging of the selective uptake of water-dispersible quantum dots into sinusoidal liver cells.

    Liang, Xiaowen; Grice, Jeffrey E; Zhu, Yian; Liu, David; Sanchez, Washington Y; Li, Zhen; Crawford, Darrell H G; Le Couteur, David G; Cogger, Victoria C; Liu, Xin; Xu, Zhi Ping; Roberts, Michael S

    2015-04-01

    Although many studies reporting the organ-level biodistribution of nanoparticles (NPs) in animals, very few have addressed the fate of NPs in organs at the cellular level. The liver appears to be the main organ for accumulation of NPs after intravenous injection. In this study, for the first time, the in vivo spatiotemporal disposition of recently developed mercaptosuccinic acid (MSA)-capped cadmium telluride/cadmium sulfide (CdTe/CdS) quantum dots (QDs) is explored in rat liver using multiphoton microscopy (MPM) coupled with fluorescence lifetime imaging (FLIM), with subcellular resolution (∼1 μm). With high fluorescence efficiency and largely improved stability in the biological environment, these QDs show a distinct distribution pattern in the liver compared to organic dyes, rhodamine 123 and fluorescein. After intravenous injection, fluorescent molecules are taken up by hepatocytes and excreted into the bile, while negatively charged QDs are retained in the sinusoids and selectively taken up by sinusoidal cells (Kupffer cells and liver sinusoidal endothelial cells), but not by hepatocytes within 3 h. The results could help design NPs targeting the specific types of liver cells and choose the fluorescent markers for appropriate cellular imaging. PMID:25504510

  4. Multiphoton spectroscopy of autoionising states and AC Stark shifts in strontium atoms

    The paper reports and discusses recent data on multiphoton ionisation of strontium, with emphasis on resonant multi photon ionisation. A theoretical framework is described for interpretation of the results, along with a description of the experiment. The results are presented as discussed with respect to the AC stark shifts of two-photon resonance, and configuration mixing for autoionising states. (U.K.)

  5. Ion beam studies of surfaces by multiphoton resonance ionization of sputtered neutrals

    Ionization of neutral atoms sputtered from ion bombarded solids by multiphoton resonance ionization has been recently demonstrated. The ionization efficiency is several orders of magnitude greater than other post-ionization methods. This approach should find applications in the characterization of the chemistry and structure of solid surfaces and in the trace analysis of a wide variety of materials. (author)

  6. Clinical combination of multiphoton tomography and high frequency ultrasound imaging for evaluation of skin diseases

    König, K.; Speicher, M.; Koehler, M. J.; Scharenberg, R.; Elsner, P.; Kaatz, M.

    2010-02-01

    For the first time, high frequency ultrasound imaging, multiphoton tomography, and dermoscopy were combined in a clinical study. Different dermatoses such as benign and malign skin cancers, connective tissue diseases, inflammatory skin diseases and autoimmune bullous skin diseases have been investigated with (i) state-of-the-art and highly sophisticated ultrasound systems for dermatology, (ii) the femtosecond-laser multiphoton tomograph DermaInspectTM and (iii) dermoscopes. Dermoscopy provides two-dimensional color imaging of the skin surface with a magnification up to 70x. Ultrasound images are generated from reflections of the emitted ultrasound signal, based on inhomogeneities of the tissue. These echoes are converted to electrical signals. Depending on the ultrasound frequency the penetration depth varies from about 1 mm to 16 mm in dermatological application. The 100-MHz-ultrasound system provided an axial resolution down to 16 μm and a lateral resolution down to 32 μm. In contrast to the wide-field ultrasound images, multiphoton tomography provided horizontal optical sections of 0.36×0.36 mm2 down to 200 μm tissue depth with submicron resolution. The autofluorescence of mitochondrial coenzymes, melanin, and elastin as well as the secondharmonic- generation signal of the collagen network were imaged. The combination of ultrasound and multiphoton tomography provides a novel opportunity for diagnostics of skin disorders.

  7. Multiphoton lonization Spectrum of Nitrogen Oxide by D~2∑←X2П

    2003-01-01

    The resonance-enhanced multiphoton ionization (REMPI) spectrum of NO has been obtained. The spectral lines can be attributed to NO D2∑←X2П transitions. NO molecules are ionized via the resonant intermediate D2∑ states and by (3+2) REMPI process.

  8. Extension of the sum rule for the transition rates between multiplets to the multiphoton case

    Solovyev, D; Volotka, A; Plunien, G

    2010-01-01

    The sum rule for the transition rates between the components of two multiplets, known for the one-photon transitions, is extended to the multiphoton transitions in hydrogen and hydrogen-like ions. As an example the transitions 3p-2p, 4p-3p and 4d-3d are considered. The numerical results are compared with previous calculations.

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

    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

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

    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

  11. Multiphoton Ionization Spectrum of Nitrogen Oxide by D2∑←X2∏

    Lianshui Zhang; Guiyin Zhang; Xiaodong Yang; Bo Sun; Xiaohui Zhao

    2003-01-01

    The resonance-enhanced multiphoton ionization (REMPI) spectrum of NO has been obtained. The spectral lines can be attributed to NO D2∑←X2∏ transitions. NO molecules are ionized via the resonant intermediate D2∑ states and by (3+2) REMPI process.

  12. Very low energy photoelectron spectroscopy in multiphoton ionization of H2

    Bordas, C.; Dyer, M.; Helm, H.

    1994-01-01

    We apply a novel photoelectron imaging spectrometer [1] to the energy and angular analysis of photoelectrons resulting from resonantly enhanced multiphoton ionization of H2. Photoelectron images with well resolved individual rotational structure of the resulting H2+ ion are obtained, demonstrating the capability of this technique for analyzing photoelectrons in the meV energy range.

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

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

    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....

  15. Resonance-enhanced multiphoton-ionization photoelectron spectroscopy of even-parity Rydberg states of atomic sulfur

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

    1996-12-01

    A (2+1) resonance-enhanced multiphoton-ionization photoelectron spectroscopy study of the sulfur atom was performed in the one-photon energy region between 260 and 240 nm. Some 20 previously unobserved even-parity Rydberg states of the sulfur atom are reported, which were accessed by two-photon transitions from the 3P ground state of the atom, prepared by in situ photodissociation of H2S. The (4So)np 3P series could be followed up to n=25. This series is perturbed around n=7 by an interloping Rydberg state converging to the first excited ionic limit 2Do. A two-channel quantum defect theory analysis was performed in order to estimate the composition of the wave functions of the perturbed series members, which is compared with the ionic state branching ratios obtained from photoelectron spectra. This analysis, moreover, enabled the determination of the ionization energy of the lowest ionic state 4So with an improved accuracy as compared to the previously reported value.

  16. Sequential multiphoton multiple ionization of Ar and Xe by X-ray free electron laser pulses at SACLA

    We have investigated multiphoton multiple ionization of Ar and Xe atoms irradiated by intense X-ray pulses using the new X-ray free electron laser facility SACLA. The experimental results are compared with theoretical results.

  17. Differential Responses of Vanilla Accessions to Root Rot and Colonization by Fusarium oxysporum f. sp. radicis-vanillae

    Koyyappurath, Sayuj; Conéjéro, Geneviève; Dijoux, Jean Bernard; Lapeyre-Montès, Fabienne; Jade, Katia; Chiroleu, Frédéric; Gatineau, Frédéric; Verdeil, Jean Luc; Besse, Pascale; Grisoni, Michel

    2015-01-01

    Root and stem rot (RSR) disease caused by Fusarium oxysporum f. sp. radicis-vanillae (Forv) is the most damaging disease of vanilla (Vanilla planifolia and V. × tahitensis, Orchidaceae). Breeding programs aimed at developing resistant vanilla varieties are hampered by the scarcity of sources of resistance to RSR and insufficient knowledge about the histopathology of Forv. In this work we have (i) identified new genetic resources resistant to RSR including V. planifolia inbreds and vanilla relatives, (ii) thoroughly described the colonization pattern of Forv into selected vanilla accessions, confirming its necrotic non-vascular behavior in roots, and (iii) evidenced the key role played by hypodermis, and particularly lignin deposition onto hypodermal cell walls, for resistance to Forv in two highly resistant vanilla accessions. Two hundred and fifty-four vanilla accessions were evaluated in the field under natural conditions of infection and in controlled conditions using in vitro plants root-dip inoculated by the highly pathogenic isolate Fo072. For the 26 accessions evaluated in both conditions, a high correlation was observed between field evaluation and in vitro assay. The root infection process and plant response of one susceptible and two resistant accessions challenged with Fo072 were studied using wide field and multiphoton microscopy. In susceptible V. planifolia, hyphae penetrated directly into the rhizodermis in the hairy root region then invaded the cortex through the passage cells where it induced plasmolysis, but never reached the vascular region. In the case of the resistant accessions, the penetration was stopped at the hypodermal layer. Anatomical and histochemical observations coupled with spectral analysis of the hypodermis suggested the role of lignin deposition in the resistance to Forv. The thickness of lignin constitutively deposited onto outer cell walls of hypodermis was highly correlated with the level of resistance for 21 accessions

  18. Characterization of the second- and third-order nonlinear optical susceptibilities of monolayer MoS$_2$ using multiphoton microscopy

    Woodward, R I; Phelan, C F; de Oliveira, R E P; Runcorn, T H; Kelleher, E J R; Li, S; de Oliveira, E C; Fechine, G J M; Eda, G; de Matos, C J S

    2016-01-01

    We report second- and third-harmonic generation in monolayer MoS$_2$ as a tool for imaging and accurately characterizing the material's nonlinear optical properties under 1560 nm excitation. Using a surface nonlinear optics treatment, we derive expressions relating experimental measurements to second- and third-order nonlinear sheet susceptibility magnitudes, obtaining values of $|\\chi_s^{(2)}|=2\\times10^{-20}$ m$^2$ V$^{-1}$ and for the first time for monolayer MoS$_2$, $|\\chi_s^{(3)}|=2\\times10^{-28}$ m$^3$ V$^{-2}$. Experimental comparisons between MoS$_2$ and graphene are also performed, demonstrating $\\sim$4 times stronger third-order nonlinearity in monolayer MoS$_2$, highlighting the material's potential for nonlinear photonics in the telecommunications C band.

  19. High-fidelity spatially resolved multiphoton counting for quantum imaging applications

    Chrapkiewicz, Radoslaw; Banaszek, Konrad

    2015-01-01

    We present a method for spatially resolved multiphoton counting based on an intensified camera with the retrieval of multimode photon statistics fully accounting for non-linearities in the detection process. The scheme relies on one-time quantum tomographic calibration of the detector. Faithful, high-fidelity reconstruction of single- and two-mode statistics of multiphoton states is demonstrated for coherent states and their statistical mixtures. The results consistently exhibit classical values of Mandel and Fano parameters in contrast to raw statistics of camera photo-events. Detector operation is reliable for illumination levels up to the average of one photon per an event area, substantially higher than in previous approaches to characterize quantum statistical properties of light with spatial resolution.

  20. On the effect of Coulomb interaction on the multiphoton ionization probability

    The nonresonant multiphoton ionization problem is considered in the case of one-dimensional Coulomb potential. The continuous spectrum wave function in the presence of electromagnetic field and Coulomb interaction is calculated in the quasiclassical approximation. The Coulomb interaction is taken into account by the use of the perturbation theory in that part of action which arises due to interaction with an electromagnetic field. Criteria of this approximation validity are found and it is shown that such an approach allows the process of nonresonant multiphoton ionization to be described in the field range εa (εa is the characteristic atomic field) for arbitrary values of the adiabaticity parameter γ. Within the range γ>>1 the Coulomb factor in the ionization probability is independing of the field strength and has to be taken into account

  1. Multiphoton ionization of the hydrogen atom exposed to circularly or linearly polarized laser pulses

    This paper studies the multiphoton ionization of the hydrogen atom exposed to the linearly or circularly polarized laser pulses by solving the time-dependent Schrödinger equation. It finds that the ratio of the ionization probabilities by linearly and circularly polarized laser pulses varies with the numbers of absorbing photons. With the same laser intensity, the circularly polarized laser pulse favors to ionize the atom with more ease than the linearly polarized laser pulse if only two or three photons are necessary to be absorbed. For the higher order multiphoton ionization, the linearly polarized laser pulse has the advantage over circularly polarized laser pulse to ionize the atom. (atomic and molecular physics)

  2. Absolute density-profile tomography of molecular beams using multiphoton ionization

    We describe an approach for the absolute density measurement of rotationally symmetric molecular beams via multiphoton ionization. This simple single-projection tomographic technique requires only knowledge of the spatial intensity profile and ionization characteristics of the focused laser beam that probes the pulsed molecular jet. Multiphoton ionization (MPI) of a xenon beam allowed tomographic reconstruction of a two-dimensional density profile with a peak density of (4.2±0.4)x1018 m-3, which was compared with the theoretical predictions of the sudden freeze model. An analytic solution to the Abel transform is derived for Gaussian projected density profiles which greatly simplifies the reconstruction of the absolute radial density. MPI is sufficiently general that this technique can be readily applied to atomic beams with a broad range of chemistries.

  3. Multiphoton ionization studies of benzene in tetramethylsilane and n-pentane solutions

    The multiphoton ionization spectra of dilute solutions (approx. -3 M) of benzene in tetramethylsilane (TMSi) and n-pentane (n-Pt) have been measured using linearly polarized light over the laser excitation wavelength (λ/sub exc/) region from 360 to 560 nm. Spectra measured with circularly polarized light are also reported for 460 <λ/sub exc/<540 nm. The order of the multiphoton ionization mechanisms that occur in different λ/sub exc/ regions has been determined, and their importance is discussed together with the observed polarization behavior. The polarization ratio and the ''apparent'' order of the ionization process were found to depend on the laser intensity. The ionization threshold of benzene in TMSi was estimated to lie between 6.6 and 5.9 eV, while in n-Pt between 7.36 and 6.8 eV

  4. Deep inner-shell multiphoton ionization by intense x-ray free-electron laser pulses

    Fukuzawa, H; Motomura, K; Mondal, S; Nagaya, K; Wada, S; Liu, X -J; Feifel, R; Tachibana, T; Ito, Y; Kimura, M; Sakai, T; Matsunami, K; Hayashita, H; Kajikawa, J; Johnsson, P; Siano, M; Kukk, E; Rudek, B; Erk, B; Foucar, L; Robert, E; Miron, C; Tono, K; Inubushi, Y; Hatsui, T; Yabashi, M; Yao, M; Santra, R; Ueda, K

    2012-01-01

    We have investigated multiphoton multiple ionization dynamics of argon and xenon atoms using a new x-ray free electron laser (XFEL) facility, SPring-8 Angstrom Compact free electron LAser (SACLA) in Japan, and identified that highly charged Xe ions with the charge state up to +26 are produced predominantly via four-photon absorption as well as highly charged Ar ions with the charge state up to +10 are produced via two-photon absorption at a photon energy of 5.5 keV. The absolute fluence of the XFEL pulse, needed for comparison between theory and experiment, has been determined using two-photon processes in the argon atom with the help of benchmark ab initio calculations. Our experimental results, in combination with a newly developed theoretical model for heavy atoms, demonstrate the occurrence of multiphoton absorption involving deep inner shells.

  5. Photoelectron circular dichroism of bicyclic ketones from multiphoton ionization with femtosecond laser pulses.

    Lux, Christian; Wollenhaupt, Matthias; Sarpe, Cristian; Baumert, Thomas

    2015-01-12

    Photoelectron circular dichroism (PECD) is a CD effect up to the ten-percent regime and shows contributions from higher-order Legendre polynomials when multiphoton ionization is compared to single-photon ionization. We give a full account of our experimental methodology for measuring the multiphoton PECD and derive quantitative measures that we apply on camphor, fenchone and norcamphor. Different modulations and amplitudes of the contributing Legendre polynomials are observed despite the similarity in chemical structure. In addition, we study PECD for elliptically polarized light employing tomographic reconstruction methods. Intensity studies reveal dissociative ionization as the origin of the observed PECD effect, whereas ionization of the intermediate resonance is dominating the signal. As a perspective, we suggest to make use of our tomographic data as an experimental basis for a complete photoionization experiment and give a prospect of PECD as an analytic tool. PMID:25492564

  6. Non-perturbative calculations for the multiphoton ionization of hydrogen and lithium atoms

    Multiphoton ionization rates for the Hydrogen atom are calculated by direct solution of the time-dependent Schrodinger equation for several intensities at a photon energy of 5.0 eV (KrF laser). Ionization rates for linear polarized light are extracted front the time evolution of the ground state on a 2d cylindrical coordinate lattice, while rates for circular polarized light are extracted from calculations on a 3d Cartesian coordinate lattice. Multiphoton ionization rates for the Lithium atom are calculated in the frozen-core TDHF approximation for a variety of intensities and photon frequencies. The time-dependent equation for the valence HF orbital is solved on a 2d cylindrical coordinate lattice using both fixed and variable grid spacings. The non-perturbative results for both atoms are in sharp contrast to perturbation theory predictions

  7. Multiphoton single ionization of two-electron systems in intense laser fields

    Multiphoton single-ionization of a two-electron system (He and He-like ions) in intense circularly polarized laser field is reported in a relativistic field theoretic method. Coulomb-correction factor is introduced to estimate the effect of the Coulomb field of the residual ion on the rate. Antisymmetric wave function is considered both in the initial and in the final state. The spin-specific currents are calculated. Angular asymmetry in current generation is noted with the change in the spin direction of the ionized electron. Coulomb-corrected relativistic-result for total rate is compared with the Coulomb-corrected nonrelativistic KFR rate. At the high-intensity regime nonrelativistic rate overestimates the relativistic rate. Formation of orthopositronium and parapositronium from positronium negative ion by multiphoton ionization is discussed.

  8. Comparison of multiphoton and collisional ionization in high intensity laser-plasma interactions

    Recent multiphoton ionization experiments, have shown that the tunneling ionization theories for complex atoms of Ammosov, et al. are valid in the regime where n* > 2 and the Keldysh tunneling parameter, γ, is less than 1. n* = Z (Eh/Eion)0.5 with Eh the ionization potential of hydrogen, and Eion that of the charge state of interest. γ = (Eion/2Φpond)0.5 where Φpond is the pondermotive potential of the laser. In high intensity laser-plasma and laser-solid interaction, tunneling ionization due to the laser field may become a dominant ionization channel. The authors discuss the laser and plasma characteristics required for multiphoton ionization to become the dominant ionization process and the effects this has on high intensity, laser-plasma and laser-solid interactions

  9. Laser-assisted multiphoton ionization of a hydrogen atom by electron impact

    Deb, S Ghosh; Sinha, C

    2008-01-01

    The dynamics of the electron impact multiphoton ionization of a hydrogen atom in the presence of an intense laser field has been studied theoretically, with a view to comparing (qualitatively) the results with the recent kinematically complete experiments of Horr et al [ Phys. Rev. Lett., vol. 94, 153201, (2005) ] for the He target. Significant laser modifications are noted in the present doubly (DDCS) and the fully differential cross sections (TDCS). For most of the explored kinematics (chosen in accordance with the experiment), the present binary peak intensity of the laser-assisted TDCS is significantly enhanced with respect to the field free ones, in agreement with the experiment but in contradiction with the existing first order theories. Importance of the multiphoton effects are also studied.

  10. High laser bandwidth single colour multiphoton ionization spectroscopy of uranium by galvanic detection

    The uranium single colour multiphoton ionization spectrum, with high laser bandwidth excitation, was measured, in the spectral range of R6G, using the fast pulsed (∝10-9s) optogalvanic signal generated by photoionization in the dark space of a hollow cathode discharge. Results indicate that this spectrum is dense in the 570 to 610 nm spectral range. A list of the 74 most intense lines is given. The results of a try at explaining these lines by a three photon resonant or quasi-resonant scheme, using known levels of uranium, are presented. The multiphoton ionization line at 591.54 nm is a special case which is discussed more thoroughly. (orig.)

  11. Multiphoton ionization (MPI) spectroscopy of tungsten hexafluoride: Experimental observation of Molecular spectrum using MPI

    In general, multiphoton ionization (MPI) spectroscopy of metal complexes shows the dissociation and ionization of the compounds. Well-structured molecular ionization spectrum is not observed yet for metal complexes by use of the multiphoton ionization technique. Tungsten hexafluoride was selected to get a molecular spectrum with MPI technique because it had a very high dissociation energy that might suppress facile photofragmentation. Also, WF6 has high vapor pressure(>1,000 torr at 300 K), which means heating is not required to get enough sample concentration in the molecular beam. The electronic absorption spectrum and the electron impact spectrum of WF6 were previously studied. The assignment of WF6 spectrum obtained by MPI can be compared with those published results. Experimentally, we observed the molecular MPI spectra of WF6 and the clear assignments are not easy with current results as well as previous works.

  12. Compact Coding Using Multi-Photon Tolerant Quantum Protocols for Quantum Communication

    Hajj, Rasha El; Chan, Kam Wai Clifford

    2016-01-01

    This paper presents a new encryption scheme called Compact Coding that encodes information in time, phase, and intensity domains, simultaneously. While these approaches have previously been used one at a time, the proposed scheme brings to bear for the first time their strengths simultaneously leading to an increase in the secure information transfer rate. The proposed scheme is applicable to both optical fibers and free space optics, and can be considered as an alternative to polarization coding. This paper applies the proposed compact coding scheme to multi-photon tolerant quantum protocols in order to produce quantum-level security during information transfer. We present the structure of the proposed coding scheme in a multi-photon environment and address its operation.

  13. In Vivo Multiphoton NADH Fluorescence Reveals Depth-Dependent Keratinocyte Metabolism in Human Skin

    Balu, Mihaela; Mazhar, Amaan; Hayakawa, Carole K.; Mittal, Richa; Krasieva, Tatiana B.; Konig, Karsten; Venugopalan, Vasan; Tromberg, Bruce J.

    2013-01-01

    We employ a clinical multiphoton microscope to monitor in vivo and noninvasively the changes in reduced nicotinamide adenine dinucleotide (NADH) fluorescence of human epidermal cells during arterial occlusion. We correlate these results with measurements of tissue oxy- and deoxyhemoglobin concentration during oxygen deprivation using spatial frequency domain imaging. During arterial occlusion, a decrease in oxyhemoglobin corresponds to an increase in NADH fluorescence in the basal epidermal c...

  14. Multiphoton-double-ionization probability linearly depends on laser intensity: Experimental studies of barium

    Bondar, I. I.; Suran, V. V.; Bondar, D. I.

    2013-01-01

    Despite inherently complex multiphoton dynamics, our observations show that Ba double ionization with an infrared laser (8800-8920 cm^{-1}) resembles a single-photon process; namely, its probability is proportional to the laser intensity. In this regime, single-electron ionization is due to a four-photon resonant transition through the highly perturbed state 6p^2 1D_2, whereas double ionization is realized by the two-electron mechanism. Furthermore, we argue that these conclusions are valid f...

  15. Study on multiphoton ionization mass spectra of 3-picoline in femtosecond laser field

    Using 800 nm femtosecond laser as excitation source, the processes and mechanism of multiphoton ionization and dissociation of 3-picoline is studied, and find the process is ionization before dissociation, the possible dissociation channels are analyzed. At B3LYP/6-311 G + + (d, p) level, we calculate the energy of every dissociation channel, the result is consistent with the laser power index of ions based on the experiment. (authors)

  16. Ionization and fragmentation of C-60 via multiphoton-multiplasmon excitation

    Hunsche, S; Starczewski, Tomas; L'Huillier, Anne; Persson, Anders; Wahlström, Claes-Göran; vandenHeuvell, B. V. L; Svanberg, Sune

    1996-01-01

    We study the intensity dependence of ionization and fragmentation of buckminsterfullerene (C-60) in strong laser fields. Our data provide strong evidence that at intensities less than or similar to 10(14) W/cm(2) these processes occur predominantly via multiphoton excitation of the 20 eV plasmon resonance of C-60 At least two plasmons have to be created to initiate fragmentation or multiple ionization.

  17. Laser-assisted multiphoton ionization of a hydrogen atom by electron impact

    Deb, S. Ghosh; S Roy; Sinha, C.

    2008-01-01

    The dynamics of the electron impact multiphoton ionization of a hydrogen atom in the presence of an intense laser field has been studied theoretically, with a view to comparing (qualitatively) the results with the recent kinematically complete experiments of Horr et al [ Phys. Rev. Lett., vol. 94, 153201, (2005) ] for the He target. Significant laser modifications are noted in the present doubly (DDCS) and the fully differential cross sections (TDCS). For most of the explored kinematics (chos...

  18. Resonance-enhanced multiphoton ionization spectroscopy oflaser-ablated copper atoms

    Andrejeva, Anna; Harris, Joe P.; Wright, Timothy G.

    2014-01-01

    Resonance-enhanced multiphoton ionization (REMPI) spectra of laser-ablated copper atoms entrainedin a supersonic free jet expansion are reported. Depending on the ionization scheme employed, andthe conditions under which the copper atoms are produced, very different spectra are produced, whichare discussed. In some circumstances, high proportions of metastable atoms survive the ablation andexpansion process and are clearly seen in the spectra. The spectroscopic transitions for the observedlin...

  19. Femtosecond time-resolved wave packet motion in molecular multiphoton ionization and fragmentation

    Baumert, Thomas,; Bühler, B; Grosser, M.; Thalweiser, Rainer; Weiss, V.; Wiedenmann, Ernst; Gerber, Gustav

    1991-01-01

    The dynamics of molecular multiphoton ionization and fragmentation of a diatomic molecule (Na_2) have been studied in molecular beam experiments. Femtosecond laser pulses from an amplified colliding-pulse mode-locked (CPM) ring dye laser are employed to induce and probe the molecular transitions. The final continuum states are analyzed by photoelectron spectroscopy, by ion mass spectrometry and by measuring the kinetic energy of the formed ionic fragments. Pump-probe spectra emplo...

  20. Alignment dependent enhancement of the photo-electron cutoff for multi-photon ionization of molecules

    Smeenk, C.T.L.; Arissian, L; Sokolov, A. V.; Spanner, M.; Lee, K F; Staudte, A.; Villeneuve, D. M.; Corkum, P B

    2013-01-01

    The multiphoton ionization rate of molecules depends on the alignment of the molecular axis with respect to the ionizing laser polarization. By studying molecular frame photo-electron angular distributions from N$_2$, O$_2$ and benzene, we illustrate how the angle-dependent ionization rate affects the photo-electron cutoff energy. We find alignment can enhance the high energy cutoff of the photo-electron spectrum when probing along a nodal plane or when ionization is otherwise suppressed. Thi...

  1. Femtosecond time-resolved molecular multiphoton ionization: the Na_2 system

    Baumert, Thomas,; Grosser, M.; Thalweiser, Rainer; Gerber, Gustav

    1991-01-01

    We report here the first experimental study of femtosecond time-resolved molecular multiphoton ionization. Femtosecond pump-probe techniques are combined with time-of-flight spectroscopy to measure transient ionization spectra of Na_2 in a molecular-beam experiment. The wave-packet motions in different molecular potentials show that incoherent contributions from direct photoionization of a singly excited state and from excitation and autoionization of a bound doubly excited molecu...

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

    2014-01-01

    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 instrumen...

  3. Studies of multiphoton ionization schemes for RIMS of La and Ba

    Multiphoton ionization (MPI) with tunable dye lasers has the potential to selectively ionize La in the presence of Ba and attain very high sensitivity. The objective of the work was to study MPI processes of Ba and La and select suitable schemes for selective ionization of La in the presence of Ba. The spectral region 520 to 610 nm was chosen because of reported strong radiative transitions in this region, and its applicability to high repetition rate Cu vapour lasers. (author)

  4. Time-resolved two-color photoacoustic and multiphoton ionization spectroscopy of aniline

    Moll, D. J.; G R Parker; Kuppermann, Aron

    1984-01-01

    The multiphoton ionization and photoacoustic signals from aniline have been detected as a function of time delay between two laser pulses of different wavelength. The first pulse excited the S0 → S1 0–0 transition. The second pulse either excited S1 to a higher singlet state, or a triplet state produced by intersystem crossing to a higher triplet state, depending on the time delay between pulses. Both ionization and dissociation processes are observed. By varying the conditions of excitation ...

  5. Resonantly enhanced multiphoton ionization and third-harmonic generation in krypton and xenon

    Multiphoton ionization (MPI) and third-harmonic generation (THG) in gaseous krypton and xenon have been studied using multiple laser photons resonant with numerous atomic energy levels of the target gas. The MPI and THG were studied concurrently in a high-pressure ionization cell with a vacuum ultraviolet (Vuv) radiation detector. Mass-resolved MPI and photoelectron spectra were recorded at low pressure (-4 torr) using the time-of-flight (TOF) mass spectrometer and double focusing spherical energy analyzer

  6. Open access

    Dorch, Bertil Fabricius; Demaio, Alessandro; Hersch, Fred

    2012-01-01

    This week, we celebrate open access week – an event aimed at bringing attention to this rapidly emerging form of scientific publication and its ethical imperatives. Traditionally, knowledge breakthroughs and scientific discoveries are shared through publication in academic journals. Peer...... ideas, break down barriers to science and make knowledge accessible to the masses – but this is not actually the case....

  7. Open Access

    Suber, Peter

    2012-01-01

    The Internet lets us share perfect copies of our work with a worldwide audience at virtually no cost. We take advantage of this revolutionary opportunity when we make our work "open access": digital, online, free of charge, and free of most copyright and licensing restrictions. Open access is made possible by the Internet and copyright-holder…

  8. Study on multiphoton ionization dissociation processes of ethyl iodide at 800 nm laser radiation

    Multiphoton ionization-dissociation (MPID) process of ethyl iodide are studied at 800 nm femto-second laser, and time-of-flight mass spectrometer of ethyl iodide are obtained. The result of laser power index shows that the parent-ion mainly undergoes the 3 + 3 resonant enhanced multiphoton ionization (REMPI) process. The percentage of part fragment ions to the total ion current and the laser intensity dependence of the molecule are measured. Based on the experiment results, the multiphoton ionization- dissociation mechanism of ethyl iodide is discussed. The conclusion is that most of ion fragments are produced by C2H5I+ dissociation, this is consistent with the parent-ion dissociation staircase. The two possible dissociation channels of parent-ion are analyzed, but the C-I dissociation is the main channel. We also use Gaussian 03 calculated the energy change of the two channels with B3LYP/3-21G basis set, the theoretical results further demonstrated the experiment. (authors)

  9. Live-cell multiphoton fluorescence correlation spectroscopy with an improved large Stokes shift fluorescent protein

    Guan, Yinghua; Meurer, Matthias; Raghavan, Sarada; Rebane, Aleksander; Lindquist, Jake R.; Santos, Sofia; Kats, Ilia; Davidson, Michael W.; Mazitschek, Ralph; Hughes, Thomas E.; Drobizhev, Mikhail; Knop, Michael; Shah, Jagesh V.

    2015-01-01

    We report an improved variant of mKeima, a monomeric long Stokes shift red fluorescent protein, hmKeima8.5. The increased intracellular brightness and large Stokes shift (∼180 nm) make it an excellent partner with teal fluorescent protein (mTFP1) for multiphoton, multicolor applications. Excitation of this pair by a single multiphoton excitation wavelength (MPE, 850 nm) yields well-separable emission peaks (∼120-nm separation). Using this pair, we measure homo- and hetero-oligomerization interactions in living cells via multiphoton excitation fluorescence correlation spectroscopy (MPE-FCS). Using tandem dimer proteins and small-molecule inducible dimerization domains, we demonstrate robust and quantitative detection of intracellular protein–protein interactions. We also use MPE-FCCS to detect drug–protein interactions in the intracellular environment using a Coumarin 343 (C343)-conjugated drug and hmKeima8.5 as a fluorescence pair. The mTFP1/hmKeima8.5 and C343/hmKeima8.5 combinations, together with our calibration constructs, provide a practical and broadly applicable toolbox for the investigation of molecular interactions in the cytoplasm of living cells. PMID:25877871

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

    The dynamics of the electron impact multiphoton ionization of a He atom in the presence of an intense laser field (nγe, 2e) is studied theoretically for laser polarization (||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)

  11. Time-resolved multiphoton imaging of basal cell carcinoma

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

    2007-02-01

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

  12. Scanning ultrafast electron microscopy

    Yang, Ding-Shyue; Mohammed, Omar F.; Zewail, Ahmed H.

    2010-01-01

    Progress has been made in the development of four-dimensional ultrafast electron microscopy, which enables space-time imaging of structural dynamics in the condensed phase. In ultrafast electron microscopy, the electrons are accelerated, typically to 200 keV, and the microscope operates in the transmission mode. Here, we report the development of scanning ultrafast electron microscopy using a field-emission-source configuration. Scanning of pulses is made in the single-electron mode, for whic...

  13. CARS microscopy for imaging

    Optical microscopy grows in its importance with the development of modern nanotechnology, biotechnology, methods of diagnostics and treatment of most dangerous diseases for mankind. There are several important goals of optical microscopy for biomedical studies among which the next three may be distinguished: fast imaging with high lateral spatial resolution, 3-D sectioning capability and high contrast for chemical selectivity. To meet these specific requirements, various types of both linear and nonlinear optical microscopy were elaborated. (authors)

  14. Nonlinear optical microscopy and microspectroscopy of oral precancers and early cancer

    Vargas, Gracie; Edward, Kert

    2013-02-01

    Multiphoton autofluorescence microscopy (MPAM) offers the ability to assess morphometry similar to that of pathologic evaluation as well as biochemical information from endogenous fluorophores which are altered with neoplastic transformation. In this study the spectroscopic properties of normal and neoplastic oral epithelium were evaluated toward the goal of identifying image/spectroscopic based indicators of neoplastic transformation using nonlinear optical microscopy. Results indicated measureable differences between normal, dysplasia, and SCC that could be helpful in delineating between the three conditions. In particular, a blue shift in autofluorescence emission was experienced for dysplasia relative to normal. However, in the case of SCC the epithelial emission experienced a significant red shift relative to both dysplasia and normal and displayed in an additional red peak that was not present in either normal or dysplastic mucosa. Results were consistent with published results for SCC in the single-photon literature. The study demonstrates that multiphoton autofluorescence spectroscopy may reveal features of oral mucosa that can be useful for differentiating normal and neoplastic mucosa. When combined with morphometry provided by MPAM, a potentially powerful technique for imaging of the oral cavity could be developed which provides both morphometric and spectroscopic information.

  15. Electron Microscopy Center (EMC)

    Federal Laboratory Consortium — The Electron Microscopy Center (EMC) at Argonne National Laboratory develops and maintains unique capabilities for electron beam characterization and applies those...

  16. Coherent light microscopy

    Ferraro, Pietro; Zalevsky, Zeev

    2011-01-01

    This book deals with the latest achievements in the field of optical coherent microscopy. While many other books exist on microscopy and imaging, this book provides a unique resource dedicated solely to this subject. Similarly, many books describe applications of holography, interferometry and speckle to metrology but do not focus on their use for microscopy. The coherent light microscopy reference provided here does not focus on the experimental mechanics of such techniques but instead is meant to provide a users manual to illustrate the strengths and capabilities of developing techniques. Th

  17. Accessible Knowledge - Knowledge on Accessibility

    Kirkeby, Inge Mette

    2015-01-01

    Although serious efforts are made internationally and nationally, it is a slow process to make our physical environment accessible. In the actual design process, architects play a major role. But what kinds of knowledge, including research-based knowledge, do practicing architects make use of when...... designing accessible environments? The answer to the question is crucially important since it affects how knowledge is distributed and how accessibility can be ensured. In order to get first-hand knowledge about the design process and the sources from which they gain knowledge, 11 qualitative interviews...... were conducted with architects with experience of designing for accessibility. The analysis draws on two theoretical distinctions. The first is research-based knowledge versus knowledge used by architects. The second is context-independent knowledge versus context-dependent knowledge. The practitioners...

  18. Open access

    Suber, Peter

    2012-01-01

    The Internet lets us share perfect copies of our work with a worldwide audience at virtually no cost. We take advantage of this revolutionary opportunity when we make our work "open access": digital, online, free of charge, and free of most copyright and licensing restrictions. Open access is made possible by the Internet and copyright-holder consent, and many authors, musicians, filmmakers, and other creators who depend on royalties are understandably unwilling to give their consent. But for 350 years, scholars have written peer-reviewed journal articles for impact, not for money, and are free to consent to open access without losing revenue. In this concise introduction, Peter Suber tells us what open access is and isn't, how it benefits authors and readers of research, how we pay for it, how it avoids copyright problems, how it has moved from the periphery to the mainstream, and what its future may hold. Distilling a decade of Suber's influential writing and thinking about open access, this is the indispe...

  19. Super-Resolved Traction Force Microscopy (STFM).

    Colin-York, Huw; Shrestha, Dilip; Felce, James H; Waithe, Dominic; Moeendarbary, Emad; Davis, Simon J; Eggeling, Christian; Fritzsche, Marco

    2016-04-13

    Measuring small forces is a major challenge in cell biology. Here we improve the spatial resolution and accuracy of force reconstruction of the well-established technique of traction force microscopy (TFM) using STED microscopy. The increased spatial resolution of STED-TFM (STFM) allows a greater than 5-fold higher sampling of the forces generated by the cell than conventional TFM, accessing the nano instead of the micron scale. This improvement is highlighted by computer simulations and an activating RBL cell model system. PMID:26923775

  20. Access French

    Grosz, Bernard

    2014-01-01

    Access is the major new language series designed with the needs of today's generation of students firmly in mind. Whether learning for leisure or business purposes or working towards a curriculum qualification, Access French is specially designed for adults of all ages and gives students a thorough grounding in all the skills required to understand, speak, read and write contemporary French from scratch. The coursebook consists of 10 units covering different topic areas, each of which includes Language Focus panels explaining the structures covered and a comprehensive glossary. Learning tips

  1. Calculations of hydrogen atom multiphoton energy level shifts, transition amplitudes and ionization probabilities

    Analyses of the resonant multiphoton ionization of atoms require knowledge of ac Stark energy shifts and of multiphoton, bound-to-bound state, transition amplitudes. In this paper, we consider the three-photon photoionization of hydrogen atoms at frequencies that are at and surrounding the two-photon 1s to 2s resonance. AC energy shift sums of both the 1s and 2s states are calculated as a function of the laser frequency along with two-photon 1s → 2s resonant transition amplitude sums. These quantities are calculated using an extended version of a method, which has often been employed in a variety of ways, of calculating these sums by expressing them in terms of solutions to a variety of differential equations that are derived from the different sums being evaluated. We demonstrate how exact solutions are obtained to these differential equations, which lead to exact evaluations of the corresponding sums. A variety of different cases are analysed, some involving analytic continuation, some involving real number analysis and some involving complex number analysis. A dc Stark sum calculation of the 2s state is carried out to illustrate the case where analytic continuation, pole isolation and pole subtraction are required and where the calculation can be carried out analytically; the 2s state, ac Stark shift sum calculations involve a case where no analytic continuation is required, but where the solution to the differential equation produces complex numbers owing to the finite photoionization lifetime of the 2s state. Results from these calculations are then used to calculate three-photon ionization probabilities of relevance to an analysis of the multiphoton ionization data published by Kyrala and Nichols (1991 Phys. Rev. A 44, R1450)

  2. Wavelength-dependent collective effects in the multiphoton ionization of atomic deuterium

    This paper presents the results of an experimental investigation into collective effects in the transient plasma formed by multiphoton ionization of atomic deuterium with a pulsed laser. The laser wavelength is varied in a narrow range around 243 nm, so that the photoionization is resonant with the metastable 2S1/2 state. The ion yield, the ion time-of-flight spectra, and the yield of Lyman-α photons have been measured as a function of laser intensity (from 1 to 340 MW/cm2) and laser detuning around the 1S1/2-2S1/2 two-photon resonance. During and shortly after the laser pulse, collective effects resulting from the mutual interaction of the photoelectrons and the ions affect the spatial and temporal distribution of the ions. Because of the near-degeneracy of the 2S1/2, 2P1/2, and 2P3/2 states, the resonant multiphoton ionization is affected by the Stark mixing of these states in the collective field. As a result, the time-dependent yields of ions and of Lyman-α photons are modulated by the interplay of the multiphoton ionization of the atoms and the collective effects in the plasma. From the measurements it is deduced that collective effects are important above a critical charge density of 3x108 ions/cm3. An asymmetry is observed in the line profile of the total ion yield as a function of laser detuning. This asymmetry is interpreted to be due to the effect of the collective field upon the intermediate resonant 2S1/2 state of the photoionization process

  3. Multiphoton ionization of a metastable hydrogen atom by electron and positron impact and charge asymmetry

    Charge asymmetry is studied theoretically for multiphoton ionization (nγ e±, e e±) of a hydrogen atom from its metastable (2S) state using coplanar geometry. The external laser field is chosen to be single-mode, spatially homogeneous, linearly polarized with laser intensity that is quite high by laboratory standards. The continuum states of the impinging electron (e) or positron (e+) are represented by plane wave Volkov states, while the wavefunctions for the ejected electron and the scattered electron/positron are chosen to be of Coulomb-Volkov type. The dressed wavefunction for the metastable (2S) hydrogen is constructed in the framework of degenerate perturbation theory. Charge asymmetry is noted in the triple differential cross sections (TDCS) of the two projectiles both in the field-free (FF) and laser-assisted situations. The positron binary peak intensity is much higher than that of the electron, while the reverse is true for the recoil peak. As for the laser modifications, the single-photon TDCS exhibits a distinct four lobed structure in contrast to the FF (two lobed) and is highly suppressed w.r.t. the latter, while the multiphoton TDCS usually tends to approach the FF (with some exceptions), obeying the famous Kroll-Watson (KW) sum rule for both projectiles. For e+ impact, the electron capture to the continuum (ECC) effect is also studied (for the symmetric geometry), where enhancement is noted in both the single and multiphoton ECC peaks thereby deviating from the KW sum rules.

  4. Time-resolved two-photon excitation fluorescence spectroscopy and microscopy using a high repetition rate streak camera

    LIU Li-xin; QU Ju-le; LIN Zi-yang; WANG Lei; FU Zhe; GUO Bao-ping; NIU Han-ben

    2007-01-01

    We present a time-resolved two-photon excitation fluorescence spectroscopy and a simultaneous time- and spectrumresolved multifocal multiphoton microscopy system that is based on a high repetition rate picosecond streak camera for providing time- and spectrum- resolved measurement and imaging in biomedicine. The performance of the system is tested and characterized by the fluorescence spectrum and lifetime analysis of several standard fluorescent dyes and their mixtures.Spectrum-resolved fluorescence lifetime images of fluorescence beads are obtained. Potential applications of the system include clinical diagnostics and cell biology etc.

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

    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.

  6. Resonant multiphoton ionization of the 1s state of a hydrogen atom in a strong laser field

    The process of resonant multiphoton ionization of hydrogen atom in the main state 1s is studied by the method of direct numerical integration of the Schroedinger nonstationary equation for quantum system in the field of an electromagnetic wave. A nonmonotonous dependence of photoionization probability on radiation intensity is found. It is ascertained that multiphoton resonances between the main and one of excited (Rydberg) atomic states induced by laser radiation field correspond to ionization minima. It is shown that ionization suppression occurs as a result of the Rydberg states restructuring in a strong field, being accompanied by effective Raman transitions binding the totality of the close-by Rydberg states

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

    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.

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

    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)

  9. Multiphoton ionization/dissociation of cyclopentanone at the lower Rydberg states

    The 2-photon excitation of the 3p and 3d Rydberg states in jet-cooled cyclopentanone has been investigated by resonance enhanced multiphoton ionization (REMPI) in a time of flight mass spectrometer. The three 3px,y,z components are clearly resolved while the case for the 3di excitations is obscure due to the S1 one-photon resonance. The ns laser induced mass spectra are characteristic of hard ionization while the fs laser induced mass spectrum is very similar to the Electron Impact one

  10. Multiphoton ionization/dissociation of cyclopentanone at the lower Rydberg states

    Philis, John G.; Kosmidis, Constantine; Tzallas, Paraskevas

    1998-12-01

    The 2-photon excitation of the 3p and 3d Rydberg states in jet-cooled cyclopentanone has been investigated by resonance enhanced multiphoton ionization (REMPI) in a time of flight mass spectrometer. The three 3px,y,z components are clearly resolved while the case for the 3di excitations is obscure due to the S1 one-photon resonance. The ns laser induced mass spectra are characteristic of hard ionization while the fs laser induced mass spectrum is very similar to the Electron Impact one.

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

    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.

  12. Spin-statistic selection rules for multiphoton transitions: Application to helium atoms

    Zalialiutdinov, T.; Solovyev, D.; Labzowsky, L.; Plunien, G.

    2016-01-01

    A theoretical investigation of the three-photon transition rates 2 1P1→2 1S0,1 1S0 and 2 3P2→2 1S0,1 1S0 for the helium atom is presented. Photon energy distributions and precise values of the nonrelativistic transition rates are obtained with employment of correlated wave functions of the Hylleraas type. The possible experiments for the tests of the Bose-Einstein statistics for multiphoton systems are discussed.

  13. Spin-Statistic Selection Rules for Multiphoton Transitions: Application to Helium Atom

    Zalialiutdinov, T; Labzowsky, L; Plunien, G

    2016-01-01

    A theoretical investigation of the three-photon transition rates $ 2^1P_1\\rightarrow 2^1S_0\\;,1^1S_0 $ and $ 2^3P_2\\rightarrow 2^1S_0\\;,1^1S_0 $ for the helium atom is presented. Photon energy distributions and precise values of the nonrelativistic transition rates are obtained with employment of correlated wave functions of the Hylleraas type. The possible experiments for the tests of the Bose-Einstein statistics for multiphoton systems are discussed.

  14. Single ionization of atoms in intense laser pulses: Evolution from multiphoton to tunnel ionization

    Rudenko, A.; Zrost, K.; Ergler, T.; Feuerstein, B.; de Jesus, V.; Schröter, C; Moshammer, R.; Ullrich, J.

    2005-01-01

    We present results of high resolution fully differential measurements on single ionization of He, Ne, and Ar by 7-25 fs linearly polarized 800nm laser pulses at intensities of up to 2.1015 W/cm2. Using a 'Reaction-Microscope' we were able to trace signatures of multiphoton ionization deep into the tunnelling regime. Surprisingly, in the low-energy electron spectra we observed several features (absence of the ponderomotive shifts, splitting of the peaks, their degeneration for few-cycle laser ...

  15. Optical-optical double-resonant multiphoton ionization spectra of Rydberg states of nitrogen dioxide

    Zhang Gui-Yin; Zhang Lian-Shui; Sun Bo; Han Xiao-Feng; Yu Wei

    2005-01-01

    The optical-optical double-resonant multiphoton ionization(OODR-MPI) technique has been applied to the study of the Rydberg states of nitrogen dioxide. The results show that ,althougy the OODR-MPI spectra of NO2 are composed of regular progression bands at different pump laser intensities, their ionization pathways are different.The NO2 mollecule is ionized through the (3+1+1)double-resonant process as the pump laser intensity is in a high value, or else it is through the (1+2+1)rpocess.The final resonant states in the two ionizing processes have been attributed to different Rydberg states.

  16. Spin-Other-Orbit Effect in Two-Mode Multiphoton Ionization

    李晓峰; 高靓辉; 王兵兵; 傅盘铭; D. S. Guo; R. R. Freeman

    2002-01-01

    A previously unknown "spin-other-orbit" effect of photon modes is predicted by solving a Schrodinger-like equation for a non-relativistic electron interacting with a two-mode intense photon field propagating in two perpendicular directions. A multiphoton ionization experiment to test this effect is suggested. Transition rates which manifest this effect to be measured in the experiment are calculated and presented graphically. A detailed comparison between this effect and the well-known spin-other-orbit effect of electrons is included.

  17. Compressor optimization with compressor-based multiphoton intrapulse interference phase scan (MIIPS).

    Hou, B; Easter, J H; Nees, J A; He, Z; Thomas, A G R; Krushelnick, K

    2012-04-15

    The multiphoton intrapulse interference phase scan (MIIPS) technique is modified to optimize the compressor settings of a chirped pulse amplification (CPA) laser system. Here, we use the compressor itself to perform the phase scan inherent in MIIPS measurement . A frequency-resolved optical gating measurement shows that the pulse duration of the compressor optimized using the modified MIIPS technique is 33.8 fs with a 2.24 rad temporal phase variation above 2% intensity. The measured time-bandwidth product is 0.60, which is close to that of transform-limited Gaussian pulse (0.44). PMID:22513694

  18. Multiphoton resonant ionization of hydrogen atom exposed to two-colour laser pulses

    Wang Pei-Jie; Fang Yan

    2008-01-01

    This paper studies the multiphoton resonant ionization by two-colour laser pulses in the hydrogen atom by solving the time-dependent Schr(o)dinger equation.By fixing the parameters of fundamental laser field and scanning the frequency of second laser field,it finds that the ionization probability shows several resonance peaks and is also much larger than the linear superposition of probabilities by applying two lasers separately.The enhancement of the ionization happens when the system is resonantly pumped to the excited states by absorbing two or more colour photons non-sequentially.

  19. Multiphoton Ionization of Formaldehyde: Observation of 3py and 3pz Rydberg States

    Bomse, D. S.; Dougal, S.

    1987-01-01

    Multiphoton ionization (MPI) of CH2O and CD2O is reported for dye laser wavelengths between 445 and 470 nm. The ionization pathway starts with three-photon resonant absorption to 3py and 3pz Rydberg states. One or two additional photons are required, depending on wavelength, to reach threshold. MPI spectra agree well with VUV absorption measurements of the same Rydberg levels. MPI yields are small and photoacoustic measurements imply the initial three-photon excitation has low probability. Tr...

  20. A NEW TECHNIQUE TO STUDY RYDBERG STATES BY MULTIPHOTON IONIZATION SPECTROSCOPY

    Verma, R.; Chanda, A.

    1987-01-01

    A new technique to study the Rydberg states of the Ba atom has been developed. In this technique a Multiphoton Ionization signal is detected by selective excitation of the ground state ion (6s) to an excited state (6p), which results in a collimated Amplified Spontaneous Emission (ASE) signal at the 6p→5d transition of Ba*. Discrete Rydberg states, 6snℓ (ℓ=0,2), as well as autoionizing Rydberg states, 5dnℓ (ℓ=0,2) and 6pnℓ (ℓ=0,2) are observed by this novel but very simple method.

  1. Resonantly Enhanced Multi-Photon Ionization Spectrum of the Neutral Green Fluorescent Protein Chromophore

    greenwood, jason; Miles, Jordan; De Camillis, Simone; Mulholland, Peter; Zhang, Lijuan; Parkes, Michael A.; Hailes, Helen C.; Fielding, Helen H.

    2014-01-01

    The photophysics of the green fluorescent protein is governed by the electronic structure of the chromophore at the heart of its β-barrel protein structure. We present the first two-color, resonance-enhanced, multiphoton ionization spectrum of the isolated neutral chromophore in vacuo with supporting electronic structure calculations. We find the absorption maximum to be 3.65 ± 0.05 eV (340 ± 5 nm), which is blue-shifted by 0.5 eV (55 nm) from the absorption maximum of the protein in its neut...

  2. Intense-field multiphoton ionization of a two-electron atom

    Multiphoton ionization of helium at high laser intensities has been investigated by direct numerical integration of the full time-dependent two-electron Schrodinger equation on a Cray T3D. At field intensities above 4 x 1014 W cm-2, two-photon ionization occurs in a few field periods, and double ionization and high harmonic generation are prominent. We present calculations of double ionization yields, investigate the sensitivity of double ionization to the electron-electron electrostatic repulsion, and present evidence of unexpected non-exponential behaviour in ionization decay rates. (Author)

  3. Two-Electron Effects in the Multiphoton Ionization of Magnesium with 400 nm 150 fs Pulses

    Xenakis, D; Karapanagioti, N. E.; Charalambidis, D.; Bachau, H.; Cormier, E

    1999-01-01

    The multiphoton ionization and photoelectron spectra of magnesium were studied at laser intensities of up to 6x10^{13} Wcm^{-2} using 150 fs laser pulses of a wavelength of 400 nm. The results indicated that a variety of different ionization mechanisms played a role in both types of spectra. A theoretical model describing the processes is presented and the routes to ionization are identified. The work demonstrates the significance of the two-electron nature of the atom in interpreting the exp...

  4. Multiphoton ionization through the triplet states of Mg by linearly and circularly polarized laser pulses

    Buica, Gabriela; 10.1103/PhysRevA.79.013419

    2013-01-01

    We theoretically study multiphoton ionization through the triplet states of Mg by linearly polarized (LP) and circularly polarized (CP) fs laser pulses. After the construction of the atomic basis using the frozen-core Hartree-Fock potential (FCHFP) as well as the model potential (MP) approaches for both singlet and triplet series which show rather good agreements with the existing data in terms of state energies and dipole matrix elements, we solve time-dependent Schr\\"{o}dinger equations with $3s3p$ $^{3}P_{1}$ as an initial state, and calculate the total ionization yield and photoelectron energy spectra (PES).

  5. The Effect of the Argon Carrier Gas in the Multiphoton Dissociation-Ionization of Tetracene

    Carmen Cisneros; Ignacio ÃÂlvarez; Alfonso Guerrero; Alejandro San Román; Juan Carlos Poveda

    2008-01-01

    The multiphoton dissociation-ionization of tetracene at 355 nm using 6.5 nanosecond laser pulses, with and without argon as a carrier gas (CG), has been studied and compared. Ion fragments were analyzed in a time-of-flight mass spectrometer and separated according to their mass-to-charge ratio (m/z). The results show that the dynamic of photodissociation at ~1010 W⋅cm-2 intensities is strongly influenced by the CG. The suppression of fragmentation channels primarily those relating to the...

  6. Self-organized pattern formation in laser-induced multiphoton ionization in fused silica

    Buschlinger, Robert; Peschel, Ulf

    2013-01-01

    We use finite difference time domain modelling to investigate plasma generation in bulk silica induced by multi-photon absorption of intense laser light. Plasma generation is found to be extremely amplified around nanometer-sized inhomogeneities as present in glasses. Each inhomogeneity acts as the seed of a plasma structure growing against the direction of light propagation. Plasma structures originating from randomly distributed inhomogeneities are found to interact strongly and to organize in regularly spaced planes oriented perpendicularly to the laser polarization. We discuss similarities between our results and nanogratings in fused silica written by laser beams with spatially homogeneous as well as radial and azimuthal polarization.

  7. Multiphoton resonant ionization of hydrogen atom exposed to two-colour laser pulses

    This paper studies the multiphoton resonant ionization by two-colour laser pulses in the hydrogen atom by solving the time-dependent Schrodinger equation. By fixing the parameters of fundamental laser field and scanning the frequency of second laser field, it finds that the ionization probability shows several resonance peaks and is also much larger than the linear superposition of probabilities by applying two lasers separately. The enhancement of the ionization happens when the system is resonantly pumped to the excited states by absorbing two or more colour photons non-sequentially. (atomic and molecular physics)

  8. Electron spectroscopy study of single and double multiphoton ionization of strontium by visible picosecond laser light

    Petite, G.; Agostini, P.

    1986-01-01

    Multiphoton single and double ionization of strontium was studied using electron spectroscopy techniques. Both a picosecond, frequency doubled Nd : Yag Laser and a picosecond rhodamine 6G Dye Laser were used, with intensities ranging from 1011 W . cm- 2 to a few 10 12 W . cm-2. Single MPI was shown to produce ions in both the ground state (3 photon) and several low lying excited states, through a four photon process. Two and three photon resonances were observed, on singly and doubly excited ...

  9. New electronic states of NH and ND observed by resonance enhanced multiphoton ionization spectroscopy

    Johnson, Russell D., III; Hudgens, Jeffrey W.

    1990-01-01

    Resonance Enhanced MultiPhoton Ionization (REMPI) spectra of NH and ND, which reveal four new electronic states are presented. Transitions from NH a 1 delta to 3s and 3p Rydberg states in both NH and ND have been observed and rotationally analyzed. The transitions were observed in the wavelength range of 258 to 288 nm. The state assignments are: e 1 pi (3s sigma) at 82857/cm, f 1 pi (3p sigma) at 86378/cm, g 1 delta (3p pi) at 88141/cm and h 1 sigma (3p pi) at 89151/cm.

  10. Multiphoton Ionization of Xenon at the LCLS Free-Electron Laser

    Rudek, B.; Rolles, D.; Rudenko, A.; Epp, S.; Foucar, L.; Erk, B.; R. Hartmann; Kimmel, N; Holl, P.; Reich, C.; Strüder, L.; Hirsemann, H.; Ueda, K.; Simon, M.; Berrah, N.

    2012-01-01

    With the first X-ray free-electron laser (FEL), the Linac Coherent Light Source (LCLS), multiphoton ionization has been pushed to a new regime, where atoms and molecules are not just ionized by a series of valence ionizations but "from the inside out". At unprecedented high intensities and short pulse durations in the soft X-ray regime, a series of inner-shell photoionizations followed by cascades of Auger decays was observed to lead to highly charged final states in rare gases such as Ne, Ar...

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

    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

  12. Direct detection of atomic ions from molecular photofragmentation during nonresonant multiphoton ionization of sputtered species

    The photoionization of sputtered Cu, Al, and Ru atoms and dimers was investigated by measuring velocity distributions using both resonant and nonresonant photoionization. Nonresonant ionization produced an atomic distribution that peaked at the same velocity as the respective dimer distribution, indicating that virtually all the nonresonant atomic ion signal is from photofragmented dimers. Various mechanisms of dimer photofragmentation are discussed. Domination of the atomic photoion channel by molecule fragmentation appears to be a general phenomenon that must be accounted for in all gas-phase multiphoton nonresonant ionization experiments at easily achievable laser power densities (≤ 109 W/cm2)

  13. Preparation of multi-photon Fock states and quantum entanglement properties in circuit QED

    We demonstrate the controllable generation of multi-photon Fock states in circuit quantum electrodynamics (circuit QED). The external bias flux regulated by a counter can effectively adjust the bias time on each superconducting flux qubit so that each flux qubit can pass in turn through the circuit cavity and thereby avoid the effect of decoherence. We further investigate the quantum correlation dynamics of coupling superconducting qubits in a Fock state. The results reveal that the lower the photon number of the light field in the number state, the stronger the interaction between qubits is, then the more beneficial to maintaining entanglement between qubits it will be. (general)

  14. Imaging NO elimination in the infrared multiphoton dissociation of nitroalkanes and alkyl nitrites

    Fernando, Ravin; Ariyasingha, Nuwandi M.; Suits, Arthur G.

    2016-02-01

    We present a DC slice imaging study of the decomposition of C2, C3 and C4 nitroalkanes and alkyl nitrites, focusing on the NO elimination channel, possibly a minor pathway. Infrared multiphoton dissociation (IRMPD) is used to induce dissociation on the ground electronic state under collisionless conditions. The channels that produced NO as a product were studied and compared among the target molecules to gain a better understanding of the isomerization of the nitroalkanes prior to dissociation. Trends in the total translational energy and NO rotational temperatures obtained from the images are discussed.

  15. Deep Inner-Shell Multiphoton Ionization by Intense X-Ray Free-Electron Laser Pulses

    Fukuzawa, H; Son, S.-K.; Kimura, M; Sakai, T; Matsunami, K; Hayashita, H.; Kajikawa, J.; Johnsson, P.; Siano, M.; Kukk, E.; Rudek, B.; Erk, B.; Motomura, K.; Foucar, L.; Robert, E.

    2013-01-01

    We have investigated multiphoton multiple ionization dynamics of argon and xenon atoms using a new x-ray free electron laser (XFEL) facility, SPring-8 Angstrom Compact free electron LAser (SACLA) in Japan, and identified that highly charged Xe ions with the charge state up to +26 are produced predominantly via four-photon absorption as well as highly charged Ar ions with the charge state up to +10 are produced via two-photon absorption at a photon energy of 5.5 keV. The absolute fluence of th...

  16. Relationship between resolution enhancement and multiphoton absorption rate in quantum lithography

    Tsang, Mankei

    2007-01-01

    The proposal of quantum lithography [Boto et al., Phys. Rev. Lett. 85, 2733 (2000)] is studied via a rigorous formalism. It is shown that, contrary to Boto et al.'s heuristic claim, the multiphoton absorption rate of a (|N,0>+|0,N>) quantum state is actually lower than that of a classical state with otherwise identical parameters. The proof-of-concept experiment of quantum lithography [D'Angelo et al., Phys. Rev. Lett. 87, 013602 (2001)] is also analyzed in terms of the proposed formalism, an...

  17. Multi-Photon Quantum Key Distribution Based on Double-Lock Encryption

    Chan, Kam Wai Clifford; Rifai, Mayssaa El; Verma, Pramode K.; Kak, Subhash; Chen, Yuhua

    2015-01-01

    This paper presents a multi-stage, multi-photon quantum key distribution protocol based on the double-lock cryptography. It exploits the asymmetry in the detection strategies between the legitimate users and the eavesdropper. The security analysis of the protocol is presented with coherent states under the intercept-resend attack, the photon number splitting attack, and the man-in-the-middle attack. It is found that the mean photon number can be much larger than one. This complements the rece...

  18. Histology in vivo: chemical contrast combined with clinical multimodal multiphoton tomography

    Weinigel, Martin; Breunig, Hans Georg; Koenig, Karsten

    2015-03-01

    Label-free multiphoton tomography based on two-photon autofluorescence, fluorescence lifetime, and second harmonic generation imaging can be supplemented by coherent anti-Stokes Raman scattering. We present a compact, mobile and flexible clinical tomograph equipped with a novel detector design with multiple miniaturized detectors for individual acquisition of all four contrast mechanisms. Imaging of endogenous fluorophores, SHG-active collagen as well as nonfluorescent lipids in human skin in vivo is possible with this clinical tomograph paving the way towards in vivo histology.

  19. Live cell imaging with chemical specificity using dual frequency CARS microscopy.

    Pope, Iestyn; Langbein, Wolfgang; Borri, Paola; Watson, Peter

    2012-01-01

    Live cell microscopy using fluorescent proteins and small fluorescent probes is a well-established and essential tool for cell biology; however, there is a considerable need for noninvasive techniques able to study tissue and cell dynamics without the need to introduce chemical or genetically encoded probes. Coherent anti-Stokes Raman scattering (CARS) microscopy is an emerging tool for cell biologists to examine live cell dynamics with chemical specificity in a label-free, noninvasive way. CARS is a multiphoton process offering intrinsic three-dimensional submicron resolution, where the image contrast is obtained from light inelastically scattered by the vibrations of endogenous chemical bonds. CARS is particularly well suited to study lipid biology, since the CARS signal of localized lipids (exhibiting a large amount of identical bonds in the focal volume) is very strong. Conversely, photostable, lipid-specific markers for fluorescence microscopy are difficult to produce and the process of labeling often affects lipid localization and function, making imaging lipids in live cells challenging, and accurate quantification often impossible. Here, we describe in detail the principles behind our experimental setup for performing CARS microscopy of lipid droplets on live cells. Since typical vibrational resonances in liquid have coherence times in the picosecond range, CARS is preferably implemented with picosecond lasers which are however expensive and less efficient than femtosecond lasers, which could also be used for other multiphoton techniques such as two-photon fluorescence. In our setup, we show that femtosecond lasers can be spectrally focused in a simple, alignment insensitive, and cost-effective way to achieve a vibrational excitation similar to picosecond lasers. This opens the way to integrate CARS and two-photon fluorescence in a single multimodal instrument for its widespread application. We also describe our dual frequency CARS system which eliminates

  20. Clinical specular microscopy

    Hirst, L.W.; Laing, R.A.

    1987-01-01

    This book provides the general ophthalmologist with a guide to the clinical applications of specular microscopy. Important material is included on laser injury, cataract surgery, corneal transplants, glaucoma, uveitis, and trauma.

  1. Magnetic microscopy guide

    Harald Brune

    2005-05-01

    Full Text Available Magnetic Microscopy of Nanostructures is an excellent introduction for newcomers and, for those working in the field, can be used as a guide before seeking more up-to-date literature, saysHarald Brune.

  2. Magnetic microscopy guide

    Harald Brune

    2005-01-01

    Magnetic Microscopy of Nanostructures is an excellent introduction for newcomers and, for those working in the field, can be used as a guide before seeking more up-to-date literature, saysHarald Brune.

  3. International Multidisciplinary Microscopy Congress

    Oral, Ahmet; Ozer, Mehmet; InterM; INTERM2013

    2014-01-01

    The International Multidisciplinary Microscopy Congress (INTERM2013) was organized on October 10-13, 2013. The aim of the congress was to bring together scientists from various branches to discuss the latest advances in the field of microscopy. The contents of the congress have been broadened to a more "interdisciplinary" scope, so as to allow all scientists working on related subjects to participate and present their work. These proceedings include 39 peer-reviewed technical papers, submitted by leading academic and research institutions from over 12 countries and representing some of the most cutting-edge research available. The 39 papers are grouped into the following sections: - Applications of Microscopy in the Physical Sciences - Applications of Microscopy in the Biological Sciences

  4. Laser diffraction microscopy

    Schall, P.

    2009-01-01

    Crystals composed of micrometer size colloidal particles diffract light and are both of fundamental interest as well as having important applications as filters, sensors and photonic devices. Laser light is used to diffract from these crystals in close analogy to x-ray or electron diffraction used for atomic crystals. Laser diffraction microscopy explores optical diffraction contrast to image crystals and crystal defects in analogy to the transmission electron microscopy technique used to ima...

  5. Interferometric synthetic aperture microscopy

    Ralston, Tyler S.; Marks, Daniel L.; Carney, P. Scott; Boppart, Stephen A.

    2007-01-01

    State-of-the-art methods in high-resolution three-dimensional optical microscopy require that the focus be scanned through the entire region of interest. However, an analysis of the physics of the light–sample interaction reveals that the Fourier-space coverage is independent of depth. Here we show that, by solving the inverse scattering problem for interference microscopy, computed reconstruction yields volumes with a resolution in all planes that is equivalent to the resolution achieved onl...

  6. Quantitative dispersion microscopy

    Fu, Dan; Choi, Wonshik; Sung, Yongjin; Yaqoob, Zahid; Dasari, Ramachandra R.; Feld, Michael

    2010-01-01

    Refractive index dispersion is an intrinsic optical property and a useful source of contrast in biological imaging studies. In this report, we present the first dispersion phase imaging of living eukaryotic cells. We have developed quantitative dispersion microscopy based on the principle of quantitative phase microscopy. The dual-wavelength quantitative phase microscope makes phase measurements at 310 nm and 400 nm wavelengths to quantify dispersion (refractive index increment ratio) of live...

  7. Multiphoton ionisation and dissociation of NO 2 by 50 fs laser pulses

    Singhal, R. P.; Kilic, H. S.; Ledingham, K. W. D.; Kosmidis, C.; McCanny, T.; Langley, A. J.; Shaikh, W.

    1996-04-01

    Multiphoton ionisation and dissociation of NO 2 has been studied experimentally at 375 nm for laser pulse widths of 10 ns and 50 fs. The parent NO 2 ion peak is not seen in the ns data. In all spectra, the main peak observed is due to the ionisation of the NO molecule which results from the dissociation of excited NO 2 formed after absorbing a 375 nm photon. The intensity dependencies of both NO and NO 2 ion peaks have also been measured. The data has been analysed within the context of a rate equation model using published cross-sections and dissociation rates except for the two-photon ionisation cross-section for NO 2 which was chosen to reproduce the NO 2/NO ion signal ratios at 50 fs. The rate equation model provides a good description of the complete set of data. Indirectly, it may be concluded that coherence effects do not play an important role in the multiphoton excitation/ionisation of NO 2. The data also rules out the importance of above-ionisation dissociation in NO 2 — a conclusion which is consistent with previous data at 496 and 248 nm for laser pulse widths ⩾ 300 fs.

  8. Secure satellite communication using multi-photon tolerant quantum communication protocol

    Darunkar, Bhagyashri; Punekar, Nikhil; Verma, Pramode K.

    2015-09-01

    This paper proposes and analyzes the potential of a multi-photon tolerant quantum communication protocol to secure satellite communication. For securing satellite communication, quantum cryptography is the only known unconditionally secure method. A number of recent experiments have shown feasibility of satellite-aided global quantum key distribution (QKD) using different methods such as: Use of entangled photon pairs, decoy state methods, and entanglement swapping. The use of single photon in these methods restricts the distance and speed over which quantum cryptography can be applied. Contemporary quantum cryptography protocols like the BB84 and its variants suffer from the limitation of reaching the distances of only Low Earth Orbit (LEO) at the data rates of few kilobits per second. This makes it impossible to develop a general satellite-based secure global communication network using the existing protocols. The method proposed in this paper allows secure communication at the heights of the Medium Earth Orbit (MEO) and Geosynchronous Earth Orbit (GEO) satellites. The benefits of the proposed method are two-fold: First it enables the realization of a secure global communication network based on satellites and second it provides unconditional security for satellite networks at GEO heights. The multi-photon approach discussed in this paper ameliorates the distance and speed issues associated with quantum cryptography through the use of contemporary laser communication (lasercom) devices. This approach can be seen as a step ahead towards global quantum communication.

  9. Role of quantum trajectory in high-order harmonic generation in the Keldysh multiphoton regime.

    Li, Peng-Cheng; Jiao, Yuan-Xiang; Zhou, Xiao-Xin; Chu, Shih-I

    2016-06-27

    We present a systematic study of spectral and temporal structure of high-order harmonic generation (HHG) by solving accurately the time-dependent Schrödinger equation for a hydrogen atom in the multiphoton regime where the Keldysh parameter is greater unity. Combining with a time-frequency transform and an extended semiclassical analysis, we explore the role of quantum trajectory in HHG. We find that the time-frequency spectra of the HHG plateau near cutoff exhibit a decrease in intensity associated with the short- and long-trajectories when the ionization process is pushed from the multiphoton regime into the tunneling regime. This implies that the harmonic emission spectra in the region of the HHG plateau near and before the cutoff are suppressed. To see the generality of this prediction, we also present a time-dependent density-functional theoretical study of the effect of correlated multi-electron responses on the spectral and temporal structure of the HHG plateau of the Ar atom. PMID:27410589

  10. Femtosecond Laser-Induced Upconversion Luminescence in Rare-Earth Ions by Nonresonant Multiphoton Absorption.

    Yao, Yunhua; Xu, Cheng; Zheng, Ye; Yang, Chengshuai; Liu, Pei; Jia, Tianqing; Qiu, Jianrong; Sun, Zhenrong; Zhang, Shian

    2016-07-21

    The upconversion luminescence of rare-earth ions has attracted considerable interest because of its important applications in photoelectric conversion, color display, laser device, multiplexed biolabeling, and security printing. Previous studies mainly explored the upconversion luminescence generation through excited state absorption, energy transfer upconversion, and photon avalanche under the continuous wave laser excitation. Here, we focus on the upconversion luminescence generation through a nonresonant multiphoton absorption by using the intense femtosecond pulsed laser excitation and study the upconversion luminescence intensity control by varying the femtosecond laser phase and polarization. We show that the upconversion luminescence of rare-earth ions under the intense femtosecond laser field excitation is easy to be obtained due to the nonresonant multiphoton absorption through the nonlinear interaction between light and matter, which is not available by the continuous wave laser excitation in previous works. We also show that the upconversion luminescence intensity can be effectively controlled by varying the femtosecond pulsed laser phase and polarization, which can open a new technological opportunity to generate and control the upconversion luminescence of rare-earth ions and also can be further extended to the relevant application areas. PMID:27367751

  11. Evidence for excitation of fluorescence in RPE melanin by multiphoton absorption

    Glickman, Randolph D.; Rockwell, Benjamin A.; Noojin, Gary D.; Stolarski, David J.; Denton, Michael L.

    2002-06-01

    Previously, we reported that ultrashort, near infrared (NIR) laser pulses caused more DNA breakage in cultured retinal pigment epithelial (RPE) cells than did CW, NIR laser radiation delivering a similar radiant exposure. We hypothesized that this difference was due to multiphoton absorption in an intracellular chromophore such as the RPE melanin. We investigated two-photon excitation of fluorescence in a suspension of isolated bovine RPE melanosomes exposed to a 1-KHz train of approximately 50- fsec laser pulses at 810 nm from a Ti:Sapphire laser, and compared this to the fluorescence excited by CW exposures at 406 nm from a Krypton ion laser. Fluorescence was measured with a PC-based spectrometer. The CW sources excited fluorescence with a peak at 525 nm. The fluorescence intensity depended on the irradiance of the sample, as well as the melanosome concentration. Peak fluorescence was obtained with a suspension of ~2 x 107 melanin granules/ml. The 810-nm, ultrashort pulses also excited fluorescence, but with a broader, lower-amplitude peak. The weaker fluorescence signal excited by the 810-nm ultrashort pulse laser for a given melanosome concentration, compared to 406-nm CW excitation, is possibly due to the smaller two- photon absorption cross-section. These results indicate the involvement of multiphoton absorption in DNA damage.

  12. Single- and Multi-Photon Events with Missing Energy in $e^+ e^-$ Collisions at LEP

    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.

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

    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, G J; Böhm, A; Boldizsar, L; Borgia, B; Bottai, S; Bourilkov, D; Bourquin, M; Braccini, S; Branson, J G; Brochu, F; Burger, J D; Burger, W J; Cai, X D; Capell, M; Carlino, G; Cartacci, A; 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, L; Cindolo, F; Clare, I; Clare, R; Coignet, G; Colino, N; Costantini, S; Cucciarelli, S; De Salvo, A; De Notaristefani, F; Debreczeni, J; Deglon, P; Degré, A; Dehmelt, K; Deiters, K; Delmeire, E; Denes, P; Diemoz, M; Dierckxsens, M; Dionisi, C; Dittmar, M; Doria, A; Dova, M T; Duchesneau, D; Duda, M; Echenard, B; El-Hage, A; El-Mamouni, H; Eline, A; 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; 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; 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; Kraber, M; Krämer, R W; Krüger, A; Kunin, A; Laktineh, I; Landi, G; Le Coultre, P; Le Goff, J M; Lebeau, M; Lebedev, A; Lebrun, P; Lecomte, P; Lecoq, P; 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 W G; 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; Napolitano, M; Natale, S; 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, F; 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; Ren, D; Rescigno, M; Reucroft, S; Riemann, S; Riles, K; Roe, B P; Romeo, G C; Romero, L; Rosca, A; Rosenbleck, C; Rosier-Lees, S; Roth, S; Rubio, J A; Ruggiero, G; Rykaczewski, H; Sakharov, A; Salicio, J; Sánchez, E; Saremi, S; Sarkar, S; Schäfer, C; Shchegelskii, V; Schopper, Herwig Franz; Schotanus, D J; Sciacca, C; Servoli, L; Shevchenko, S; Shivarov, N; Shoutko, V; Shumilov, E; Shvorob, A; 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, K; Swain, J D; Szillási, Z; Tang, X W; Tarjan, P; Tauscher, L; 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, G; 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; De Asmundis, R; De Guevara, P L; de Jong, P; de la Cruz, B; Della Volpe, D; van Dalen, J A; van Gulik, R

    2004-01-01

    Single- and multiphoton events with missing energy are selected in 619 pb/sup -1/ of data collected by the L3 detector at LEP at centre- of-mass energies between 189 and 209 GeV. The cross sections of the process e^{+}e^{-} to 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/sub nu /=2.98+or-0.05+or-0.04. Selection results are given in the form of tables which can be used to test future models involving single- and multiphoton 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 1.5 and 0.65 TeV are set, at 95% confidence level, on the new scale of gravity for the number of extra dimensions between 2 and 6.

  14. Characterization of multiphoton photoacoustic spectroscopy for subsurface brain tissue diagnosis and imaging

    Dahal, Sudhir; Cullum, Brian M.

    2016-04-01

    The development and demonstration of a multiphoton photoacoustic imaging technique capable of providing high spatial resolution chemical images of subsurface tissue components as deep as 1.4 cm below the tissue surface is described. By combining multiphoton excitation in the diagnostic window (650 to 1100 nm), with ultrasonic detection of nonradiative relaxation events, it is possible to rapidly reconstruct three-dimensional, chemical specific, images of samples underneath overlying structures as well as chemical species of the same material. Demonstration of this technique for subsurface tissue differentiation is shown, with the ability to distinguish between grade III astrocytoma tissue and adjacent healthy tissue in blind studies. By employing photoacoustic signal detection, the high nonradiative relaxation rates of most biological tissue components (>90%) and the minimal signal attenuation of the resulting ultrasound compensate for excitation efficiency losses associated with two-photon absorption. Furthermore, the two-photon absorption process results in a highly localized excitation volume (ca., 60 μm). Characterization of the probing depth, spatial resolution, and ability to image through overlying structures is also demonstrated in this paper using tissue phantoms with well-characterized optical scattering properties, mimicking those of tissues.

  15. Development and characterization of non-resonant multiphoton photoacoustic spectroscopy (NMPPAS) for brain tumor margining

    Dahal, Sudhir

    During tumor removal surgery, due to the problems associated with obtaining high-resolution, real-time chemical images of where exactly the tumor ends and healthy tissue begins (tumor margining), it is often necessary to remove a much larger volume of tissue than the tumor itself. In the case of brain tumor surgery, however, it is extremely unsafe to remove excess tissue. Therefore, without an accurate image of the tumor margins, some of the tumor's finger-like projections are inevitably left behind in the surrounding parenchyma to grow again. For this reason, the development of techniques capable of providing high-resolution real-time images of tumor margins up to centimeters below the surface of a tissue is ideal for the diagnosis and treatment of tumors, as well as surgical guidance during brain tumor excision. A novel spectroscopic technique, non-resonant multiphoton photoacoustic spectroscopy (NMPPAS), is being developed with the capabilities of obtaining high-resolution subsurface chemical-based images of underlying tumors. This novel technique combines the strengths of multiphoton tissue spectroscopy and photoacoustic spectroscopy into a diagnostic methodology that will, ultimately, provide unparalleled chemical information and images to provide the state of sub-surface tissues. The NMPPAS technique employs near-infrared light (in the diagnostic window) to excite ultraviolet and/or visible light absorbing species deep below the tissue's surface. Once a multiphoton absorption event occurs, non-radiative relaxation processes generates a localized thermal expansion and subsequent acoustic wave that can be detected using a piezoelectric transducer. Since NMPPAS employs an acoustic detection modality, much deeper diagnoses can be performed than that is possible using current state of the art high-resolution chemical imaging techniques such as multiphoton fluorescence spectroscopy. NMPPAS was employed to differentiate between excised brain tumors (astrocytoma III

  16. Quantitative evaluation of the effect of multiphoton ionization in laser Thomson scattering diagnostics of low-temperature plasmas

    Kono, Akihiro; Matsuda, Yukitaka; Okada, Ken; Aramaki, Mitsutoshi

    2011-10-01

    Laser Thomson scattering technique allows one very high spatial-resolution measurements of electron density and temperature as required in diagnostics of microplasmas or measurement of regions very close to the wall. In such high-resolution measurements, laser energy density in the scattering volume must be very high for obtaining sufficient amount of scattering photons; therefore, electron production via multiphoton ionization by the laser beam may significantly affect Thomson scattering due to plasma electrons. In order to clarify the effect of multiphoton ionization in Thomson scattering diagnostics, we have been measuring absolute efficiency of multiphoton ionization caused by frequency-doubled Nd:YAG laser for various gases including rare gases, N2, O2, CF4 and SF6. It has been found that electrons produced via multiphoton ionization can reach a significant fraction of plasma electrons even at low pressures, depending on the gas species and laser focusing conditions. Details will be given in the presentation, including simulation results about how the escape of photoelectrons from the scattering volume affects the situation.

  17. “Single Shot” Laser Multiphoton Ionization Detection of UF5 Following the 266 nm Photodissociation of UF6

    Chou, J. S.; Sumida, D.; Stuke, M.; Wittig, C

    1982-01-01

    By using laser multiphoton ionization and a time-of-flight mass filter, we have detected nascent UF5 from the 266 nm laser photodissociation of UF6, following a single laser firing. The method can be used to monitor these species under conditions wherein isotopically selective excitation is responsible for the dissociation of UF6.

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

    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...

  19. Verification Results of Jet Resonance-enhanced Multiphoton Ionization as a Real-time PCDD/F Emission Monitor

    The Jet REMPI (Resonance Enhanced Multiphoton Ionization) monitor was tested on a hazardous waste firing boiler for its ability to determine concentrations of polychlorinated dibenzodioxins and dibenzofurans (PCDDs/Fs). Jet REMPI is a real time instrument capable of highly selec...

  20. Interferometric synthetic aperture microscopy

    Ralston, Tyler S.; Marks, Daniel L.; Scott Carney, P.; Boppart, Stephen A.

    2007-02-01

    State-of-the-art methods in high-resolution three-dimensional optical microscopy require that the focus be scanned through the entire region of interest. However, an analysis of the physics of the light-sample interaction reveals that the Fourier-space coverage is independent of depth. Here we show that, by solving the inverse scattering problem for interference microscopy, computed reconstruction yields volumes with a resolution in all planes that is equivalent to the resolution achieved only at the focal plane for conventional high-resolution microscopy. In short, the entire illuminated volume has spatially invariant resolution, thus eliminating the compromise between resolution and depth of field. We describe and demonstrate a novel computational image-formation technique called interferometric synthetic aperture microscopy (ISAM). ISAM has the potential to broadly impact real-time three-dimensional microscopy and analysis in the fields of cell and tumour biology, as well as in clinical diagnosis where in vivo imaging is preferable to biopsy.

  1. Following intracellular cholesterol transport by linear and non-linear optical microscopy of intrinsically fluorescent sterols.

    Wüstner, Daniel

    2012-02-01

    Elucidation of intracellular cholesterol transport is important for understanding the molecular basis of several metabolic and neuronal diseases, like atheroclerosis or lysosomal storage disorders. Progress in this field depends crucially on the development of new technical approaches to follow the cellular movement of this essential lipid molecule. In this article, a survey of the various methods being used for analysis of sterol trafficking is given. Various classical biochemical methods are presented and their suitability for analysis of sterol trafficking is assessed. Special emphasis is on recent developments in imaging technology to follow the intracellular fate of intrinsically fluorescent sterols as faithful cholesterol markers. In particular, UV-sensitive wide field and multiphoton microscopy of the sterol dehydroergosterol, DHE, is explained and new methods of quantitative image analysis like pixel-wise bleach rate fitting and multiphoton image correlation spectroscopy are introduced. Several applications of the new technology including observation of vectorial sterol trafficking in polarized human hepatoma cells for investigation of reverse cholesterol transport are presented. PMID:21470123

  2. Confocal Raman Microscopy

    Dieing, Thomas; Toporski, Jan

    2011-01-01

    Confocal Raman Microscopy is a relatively new technique that allows chemical imaging without specific sample preparation. By integrating a sensitive Raman spectrometer within a state-of-the-art microscope, Raman microscopy with a spatial resolution down to 200nm laterally and 500nm vertically can be achieved using visible light excitation. Recent developments in detector and computer technology as well as optimized instrument design have reduced integration times of Raman spectra by orders of magnitude, so that complete images consisting of tens of thousands of Raman spectra can be acquired in seconds or minutes rather than hours, which used to be standard just one decade ago. The purpose of this book is to provide the reader a comprehensive overview of the rapidly developing field of Confocal Raman Microscopy and its applications.

  3. Controllable tomography phase microscopy

    Xiu, Peng; Zhou, Xin; Kuang, Cuifang; Xu, Yingke; Liu, Xu

    2015-03-01

    Tomography phase microscopy (TPM) is a new microscopic method that can quantitatively yield the volumetric 3D distribution of a sample's refractive index (RI), which is significant for cell biology research. In this paper, a controllable TPM system is introduced. In this system a circulatory phase-shifting method and piezoelectric ceramic are used which enable the TPM system to record the 3D RI distribution at a more controllable speed, from 1 to 40 fps, than in the other TPM systems reported. The resolution of the RI distribution obtained by this controllable TPM is much better than that in images recorded by phase contrast microscopy and interference tomography microscopy. The realization of controllable TPM not only allows for the application of TPM to the measurement of kinds of RI sample, but also contributes to academic and technological support for the practical use of TPM.

  4. Magnetic force microscopy

    Principles of the operation of the magnetic force microscope are considered. The main areas of application of magnetic force microscopy are characterised. The prospects are shown of using this method for solving problems of materials science, including the most topical problem, viz., the creation of magnetic recording media with ultrahigh recording density. Unique potentials for investigation of nanostructures using a combination of magnetic force microscopy with electron spin resonance and nuclear magnetic resonance are pointed out. Primary attention is focused on the description of experimental techniques. The bibliography includes 27 references.

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

    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

  6. When Is Open Access Not Open Access?

    MacCallum, Catriona J.

    2007-01-01

    As open access grows in prominence, so too has confusion about what open access means; such confusion arises from a genuine misunderstanding of open access by funders, authors, editors, and publishers alike.

  7. Yeast fluorescence microscopy

    Hašek, Jiří

    New Jersey : Humana Press, 2005, s. 85-96. ISBN 1-59259-958-3 R&D Projects: GA AV ČR IAA5020102; GA ČR GA204/02/1424 Institutional research plan: CEZ:AV0Z50200510 Keywords : yeast * fluorescence microscopy * immunofluorescence Subject RIV: EE - Microbiology, Virology

  8. Dynamic saturation optical microscopy

    Sýkora, Jan; Dertinger, T.; Enderlein, J.

    Berlin : Max Delbrück Center for Molecular Medicine, 2006 - (Cardoso, M.). s. 56 [International Symposium Optical Analysis of Biomolecular Machines. 13.07.2006-16.07.2006, Berlin] Institutional research plan: CEZ:AV0Z40400503 Keywords : optical microscopy * physical chemistry Subject RIV: CF - Physical ; Theoretical Chemistry

  9. Correlative Stochastic Optical Reconstruction Microscopy and Electron Microscopy

    Kim, Doory; Deerinck, Thomas J.; Sigal, Yaron M.; Babcock, Hazen P.; Ellisman, Mark H.; Zhuang, Xiaowei

    2015-01-01

    Correlative fluorescence light microscopy and electron microscopy allows the imaging of spatial distributions of specific biomolecules in the context of cellular ultrastructure. Recent development of super-resolution fluorescence microscopy allows the location of molecules to be determined with nanometer-scale spatial resolution. However, correlative super-resolution fluorescence microscopy and electron microscopy (EM) still remains challenging because the optimal specimen preparation and ima...

  10. Scanning Probe Microscopy and Spectroscopy

    Wiesendanger, Roland

    1994-09-01

    Preface; List of acronyms; Introduction; Part I. Experimental Methods and Theoretical Background of Scanning Probe Microscopy and Spectroscopy: 1. Scanning tunnelling microscopy; 2. Scanning force microscopy; 3. Related scanning probe techniques; Part II. Applications of Scanning Probe Microscopy and Spectroscopy: 4. Condensed matter physics; 5. Chemistry; 6. Organic materials; 7. Metrology and standards; 8. Nanotechnology; References; Index.

  11. Flame temperature measurements by radar resonance-enhanced multiphoton ionization of molecular oxygen.

    Wu, Yue; Sawyer, Jordan; Zhang, Zhili; Adams, Steven F

    2012-10-01

    Here we report nonintrusive local rotational temperature measurements of molecular oxygen, based on coherent microwave scattering (radar) from resonance-enhanced multiphoton ionization (REMPI) in room air and hydrogen/air flames. Analyses of the rotational line strengths of the two-photon molecular oxygen C(3)Π(v=2)←X(3)Σ(v'=0) transition have been used to determine the hyperfine rotational state distribution of the ground X(3)Σ(v'=0) state. Rotationally resolved 2+1 REMPI spectra of the molecular oxygen C(3)Π(v=2)←X(3)Σ(v'=0) transition at different temperatures were obtained experimentally by radar REMPI. Rotational temperatures have been determined from the resulting Boltzmann plots. The measurements in general had an accuracy of ~±60 K in the hydrogen/air flames at various equivalence ratios. Discussions about the decreased accuracy for the temperature measurement at elevated temperatures have been presented. PMID:23033104

  12. Laser and molecules: the season of multiphoton in Frascati in the years 70 and 80

    This paper consists of a historical review of the research activities going on at the end of the ‘70 and beginning of the ’80 at ENEA Frascati Research Centre on the topic of infrared laser molecule interactions in the gas phase. Main goal of the studies was the implementation of isotopically selective multiphoton dissociation. The activity, conducted in an extended framework of fruitful international cooperation, ranged from fundamental research addressed to the understanding of the mechanism of the excitation process in climbing the vibro-rotational ladder, to applicative consequences in the realization of process prototypes (laser, reactors) suitable to successive commercialization. The entire activity successively pushed the laboratory development in the ’90 both on fundamental research themes (like the high resolution IR spectroscopy and the study of molecular clusters in ultracold supersonic beams) and on other applicative fields, like laser driven nano-technologies and lidar systems

  13. Aromatic C-H bond activation revealed by infrared multiphoton dissociation spectroscopy.

    Jašíková, Lucie; Hanikýřová, Eva; Schröder, Detlef; Roithová, Jana

    2012-04-01

    Metal-oxide cations are models of catalyst mediating the C-H bond activation of organic substrates. One of the most powerful reagents suggested in the gas phase is based on CuO(+) . Here, we describe the activation of the aromatic C-H bonds of phenanthroline in its complex with CuO(+) . The reaction sequence starts with a hydrogen atom abstraction by the oxygen atom from the 2-position of the phenanthroline ring, followed by OH migration to the ring. Using infrared multiphoton spectroscopy, it is shown that the reaction can be energetically facilitated by additional coordination of a water ligand to the copper ion. As the reaction is intramolecular, a spectroscopic characterization of the product is mandatory in order to unambiguously address the reaction mechanism. PMID:22689621

  14. Resonance-Enhanced Multiphoton Ionization Mass Spectrometry (REMPI-MS): Applications for Process Analysis

    Streibel, Thorsten; Zimmermann, Ralf

    2014-06-01

    Process analysis is an emerging discipline in analytical sciences that poses special requirements on analytical techniques, especially when conducted in an online manner. Mass spectrometric methods seem exceedingly suitable for this task, particularly if a soft ionization method is applied. Resonance-enhanced multiphoton ionization (REMPI) in combination with time-of-flight mass spectrometry (TOFMS) provides a selective and sensitive means for monitoring (poly)aromatic compounds in process flows. The properties of REMPI and various variations of the ionization process are presented. The potential of REMPI for process analysis is highlighted with several examples, and drawbacks of the method are also noted. Applications of REMPI-TOFMS for the detection and monitoring of aromatic species in a large variety of combustion processes comprising flames, vehicle exhaust, and incinerators are discussed. New trends in technical development and combination with other analytical methods are brought forward.

  15. Engineering entangled microwave photon states through multiphoton interactions between two cavity fields and a superconducting qubit

    Zhao, Yan-Jun; Wang, Changqing; Zhu, Xiaobo; Liu, Yu-xi

    2016-01-01

    It has been shown that there are not only transverse but also longitudinal couplings between microwave fields and a superconducting qubit with broken inversion symmetry of the potential energy. Using multiphoton processes induced by longitudinal coupling fields and frequency matching conditions, we design a universal algorithm to produce arbitrary superpositions of two-mode photon states of microwave fields in two separated transmission line resonators, which are coupled to a superconducting qubit. Based on our algorithm, we analyze the generation of evenly-populated states and NOON states. Compared to other proposals with only single-photon process, we provide an efficient way to produce entangled microwave photon states when the interactions between superconducting qubits and microwave fields are in the strong and ultrastrong regime. PMID:27033558

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

    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.)

  17. Optical properties of multicoated CdSe/CdS/ZnS quantum dots for multiphoton applications

    CdSe-core CdS/Cd0.5Zn0.5S/ZnS multishell quantum dots (QDs) were synthesized by using the successive ion layer adsorption and reaction method, and their optical properties were characterized experimentally. Based on probability calculations of the time-dependent Schroedinger equation, a kinetic Monte Carlo method was used to simulate and analyse the absorption and spontaneous emission spectra of multiphoton-excited QDs. It has been shown that the blue shift of the photoluminescence peak of the core CdSe QD after coating is most probably due to the Zn diffusion during the synthesis of the ZnS coating layer

  18. Multi-Photon Multi-Channel Interferometry for Quantum Information Processing

    Dhand, Ish

    2016-01-01

    This thesis reports advances in the theory of design, characterization and simulation of multi-photon multi-channel interferometers. I advance the design of interferometers through an algorithm to realize an arbitrary discrete unitary transformation on the combined spatial and internal degrees of freedom of light. This procedure effects an arbitrary $n_{s}n_{p}\\times n_{s}n_{p}$ unitary matrix on the state of light in $n_{s}$ spatial and $n_{p}$ internal modes. I devise an accurate and precise procedure for characterizing any multi-port linear optical interferometer using one- and two-photon interference. Accuracy is achieved by estimating and correcting systematic errors that arise due to spatiotemporal and polarization mode mismatch. Enhanced accuracy and precision are attained by fitting experimental coincidence data to a curve simulated using measured source spectra. The efficacy of our characterization procedure is verified by numerical simulations. I develop group-theoretic methods for the analysis and ...

  19. Influence of laser intensity on the double-resonance multiphoton ionization process of NO molecule

    Guiyin Zhang; Yidong Jin

    2008-01-01

    The analytic forrnula of the ionization efficiency in the process of double resonance enhanced multi-photon ionization (DREMPI) is derived from the dynamic rate equation about the interaction of photon and material. Based on rids formula, the ionization efficiency arid the laser power index versus laser intensity in the DREMPI process of NO molecule, via A2∑ and S2∑ intermediate resonant states, is numerically sinnllated. It is shown that the ionization efficiency of NO molecule increases with the laser intensity until getting saturation, while the laser power index decreases with the enhancement of the laser intensity arid changes to zero at last. The variation of the laser power index with the laser intensity indicates that the ionization efficiency reaches saturation in the one, two, and three excitation steps respectively. It is also found that the narrower the laser pulse duration is, the higher becomes the laser intensity for saturation.

  20. Superresolving Imaging of Irregular Arrays of Thermal Light Sources using Multiphoton Interferences

    Classen, Anton; Giebel, Sebastian; Schneider, Raimund; Bhatti, Daniel; Mehringer, Thomas; von Zanthier, Joachim

    2016-01-01

    We propose to use multiphoton interferences of photons emitted from statistically independent thermal light sources in combination with linear optical detection techniques to reconstruct, i.e., image, arbitrary source geometries in one dimension with subclassical resolution. The scheme is an extension of earlier work [Phys. Rev. Lett. 109, 233603 (2012)] where N regularly spaced sources in one dimension were imaged by use of the Nth-order intensity correlation function. Here, we generalize the scheme to reconstruct any number of independent thermal light sources at arbitrary separations in one dimension exploiting intensity correlation functions of order $m \\geq 3$. We present experimental results confirming the imaging protocol and provide a rigorous mathematical proof for the obtained subclassical resolution.

  1. Chiral asymmetry in the multiphoton ionization of methyloxirane using femtosecond electron-ion coincidence imaging.

    Rafiee Fanood, Mohammad M; Powis, Ivan; Janssen, Maurice H M

    2014-12-11

    Multiphoton photoelectron circular dichroism (MP-PECD) has been observed as an asymmetry in the angular distribution of photoelectrons emitted in the ionization of pure enantiomers of the small chiral molecule methyloxirane using a femtosecond laser operated at 420 nm. Energetically, this requires the uptake of four photons. By switching the laser between left- and right-circular polarization, and observing the differences in the full three-dimensional electron momentum distribution recorded in an electron-ion coincidence technique, the chiral (odd) terms in the angular distribution expression can be isolated. Electron events can additionally be filtered by coincident ion mass, providing mass-tagged electron distributions and quantitative measures of the MP-PECD asymmetry that help characterize the different ionization channels. For the production of ground state parent cation, the magnitude of the mean chiral asymmetry is measured to be 4.7%, with a peak magnitude exceeding 10% PMID:25402546

  2. Multiphoton ionization and dissociation of CH3I at 266 and 355 nm

    Li Li; Xianghe Kong; Shudong Zhang

    2007-01-01

    The mechanisms of multiphoton ionization (MPI) and dissociation of CH3I have been studied using timeof-flight (TOF) mass spectrometer at 266 and 355 nm. MPI mass spectrum at 266 nm consists mostly of fragment ions. This is consistent with a neutral-fragment photoionization mechanism in which rapid one-photon dissociation occurs from the repulsive potential energy surface followed by MPI of neutral photofragments. The observation of parent ions at 355-nm excitation is indicative of parent-ionic ladder mechanism in which the parent ions are produced directly by two-photon excitation resonantly excited to Rydberg C state and then ionized through additional one-photon absorption to produce CH3I+. Fragment ions are produced by dissociation of CH3I+.

  3. A time-dependent variational approach to multiphoton ionization of H atoms in intense laser fields

    We present calculations using a variational method for the time-dependent Schrodinger equation for the study of multiphoton ionization of H atoms in intense laser fields. The trial wavefunction is chosen to be an anisotropic Gaussian wavepacket and the case of linear polarization of the laser field is considered. We report on ionization rates as a function of laser intensity (in the range 1015-1016W cm2) and frequency (corresponding to ionization by three or more photons) and momentum-dependent electron spectra. Comparison is made with results obtained when the trial wavefunction is an isotropic Gaussian wavepacket and with large-scale numerical calculations. The present method gives reliable results for non-resonant ionization in the limit of high field intensity as well as in the low-frequency limit. The electron momentum distributions are singly peaked and provide a qualitative picture of the ionization process at high field strengths. (author)

  4. Electron impact ionization and multiphoton ionization of doped superfluid helium droplets: A comparison.

    He, Yunteng; Zhang, Jie; Kong, Wei

    2016-02-28

    We compare characteristics of electron impact ionization (EI) and multiphoton ionization (MPI) of doped superfluid helium droplets using the same droplet source. Selected dopant ion fragments from the two ionization schemes demonstrate different dependence on the doping pressure, which could be attributed to the different ionization mechanisms. While EI directly ionizes helium atoms in a droplet therefore has higher yields for bigger droplets (within a limited size range), MPI is insensitive to the helium in a droplet and is only dependent on the number of dopant molecules. The optimal timing of the ionization pulse also varies with the doping pressure, implying a velocity slip among different sized droplets. Calculations of the doping statistics and ionization probabilities qualitatively agree with the experimental data. Our results offer a word of caution in interpreting the pressure and timing dependence of superfluid helium droplets, and we also devise a scheme in achieving a high degree of doping while limiting the contribution of dopant clusters. PMID:26931697

  5. Coulomb asymmetry and sub-cycle electron dynamics in multiphoton multiple ionization of H2

    We present a systematic study of the molecular-frame photo-electron angular distributions produced by multiphoton double ionization of H2 using circularly polarized 800 nm, femtosecond laser pulses. We compare experimental results to numerical results obtained from a reduced-dimensionality time-dependent Schrödinger equation (TDSE) model. In addition, we implement a TDSE-like version of the strong-field approximation to isolate the effect of the parent ion’s Coulomb potential on the continuum electron in our simulations. Thereby we identify the contributions of the parent ion potential, and light induced sub-optical cycle electron dynamics on the observable energy and angular distributions.

  6. Resonant and nonresonant multiphoton ionization processes in the mass spectrometry of explosives.

    Hamachi, Akifumi; Okuno, Tomoya; Imasaka, Tomoko; Kida, Yuichiro; Imasaka, Totaro

    2015-03-01

    Multiphoton ionization processes were studied for three types of explosives using a line-tunable ultraviolet femtosecond laser. When peroxides such as triacetone triperoxide (TATP) and hexamethylene triperoxide diamine (HMTD) were ionized through a nonresonant two-photon process, a molecular ion was dominantly observed by reducing the excess energy remaining in the ion. However, an aromatic nitro compound such as 2,4,6-trinitrotoluene (TNT) produced large signals arising from molecular and fragment ions by resonant two-photon ionization. In addition, only fragment ions were produced from a nonaromatic nitro compound such as 1,3,5-trinitroperhydro-1,3,5-triazine (RDX), even when a resonant two-photon ionization process was employed, suggesting that a further reduction in excess energy would be necessary if a molecular ion were to be observed. PMID:25622138

  7. The Effect of the Argon Carrier Gas in the Multiphoton Dissociation-Ionization of Tetracene

    Carmen Cisneros

    2008-10-01

    Full Text Available The multiphoton dissociation-ionization of tetracene at 355 nm using 6.5 nanosecond laser pulses, with and without argon as a carrier gas (CG, has been studied and compared. Ion fragments were analyzed in a time-of-flight mass spectrometer and separated according to their mass-to-charge ratio (m/z. The results show that the dynamic of photodissociation at ~1010 W⋅cm-2 intensities is strongly influenced by the CG. The suppression of fragmentation channels primarily those relating to the formation of the CHm+ (m = 2, 4, C2H4+ and C5H4+2 ions. CH5+ and CH6+ were observed which have not been reported before in photodissociation tetracene experiments.

  8. Multiphoton Ionization Time-of-Flight Mass Spectrometry for the Detection of Bioactive Lignan.

    Uchimura, Tomohiro; Tokumoto, Goro; Batnyam, Onon; Chou, Chih-Wei; Fujita, Satoshi

    2016-01-01

    Multiphoton ionization time-of-flight mass spectrometry (MPI-TOFMS) combined with a pulsed laser for sample vaporization was developed for the detection of a low-volatile compound in a solution. A solution containing Taiwanin A ((3E,4E)-3,4-bis(1,3-benzodioxol-5-ylmethylene)dihydro-2(3H)-furanone), which is a lignan that has an anticancer effect, was employed in the present study. Consequently, Taiwanin A could be detected by irradiating a laser pulse for vaporization to an inlet nozzle, rather than by heating. Therefore, the present method could be effective for detecting compounds with lower volatilities in a liquid sample. PMID:26860576

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

    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.

  10. Spatiotemporal steering of photoelectron emission in multiphoton above-threshold ionization

    Gong, Xiaochun; Liu, Mingming; Song, Qiying; Lin, Kang; Ji, Qinying; Zhang, Wenbin; Ma, Junyang; Lu, Peifen; Liu, Yunquan; He, Feng; Zeng, Heping; Yang, Weifeng; Wu, Jian

    2016-01-01

    We experimentally demonstrate spatiotemporal steering of photoelectron emission in multiphoton above-threshold single ionization of atoms exposed to a phase-controlled orthogonally polarized two-color (OTC) laser pulse. Spatial and energy resolved photoelectron angular distributions are measured as a function of the laser phase, allowing us to look into the fine structures and emission dynamics. The slow and fast photoelectrons, distinguished by the energy larger or smaller than 2Up with Up being the ponderomotive energy of a free electron in the laser field, have distinct spatiotemporal dependences of the laser waveform. The phase-of-phase of the slow electron oscillates as functions of both the energy and emission direction, however, the fast electron present rather flat phase structure which merely depends on its emission direction. Three-dimensional generalized quantum trajectory Monte Carlo simulations are performed to explore the sub-cycle dynamics of the electron emission in the phase-controlled OTC pu...

  11. Three-Dimensional Organic Microlasers with Low Lasing Thresholds Fabricated by Multiphoton Lithography

    Chen, Vincent W; Lafargue, Clement; Mansfield, Eric S; Yom, Jeannie; Johnstone, Luke; Hales, Joel M; Bittner, Stefan; Charpignon, Severin; Ulbricht, David; Lautru, Joseph; Denisyuk, Igor; Zyss, Joseph; Perry, Joseph W; Lebental, Melanie

    2014-01-01

    Cuboid-shaped organic microcavities containing a pyrromethene laser dye and supported upon a photonic crystal have been investigated as an approach to reducing the lasing threshold of the cavities. Multiphoton lithography facilitated fabrication of the cuboid cavities directly on the substrate or on the decoupling structure, while similar structures were fabricated on the substrate by UV lithography for comparison. Significant reduction of the lasing threshold by up to a factor of 4 has been observed for cavities supported by the photonic crystal relative to those fabricated on the substrate. The lasing mode spectra of the cuboid microresonators provide strong evidence showing that the lasing modes are localized in the horizontal plane, with the shape of an inscribed diamond.

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

    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 A2 term and the corresponding orders of the multipolar expansion of the transition matrix.

  13. Entanglement of molecular-orientation, rotational and orbital degrees of freedom in multiphoton orientational wave packets

    Mainos, C.; Dutier, G.; Grucker, J.; Perales, F.; Baudon, J.; Ducloy, M.

    2008-02-01

    Multiphoton orientational wave packets induced by short resonant polarized laser pulses in a rotationally-frozen interacting molecule contain relevant information. The entanglement of the orbital, rotational and orientational degrees of freedom shows a strong dependence on the polarization state of the absorbed photons and the space orientation of the interacting molecule and enables one to assign the orbital state of the excited molecular electron, to measure the space orientation of the interacting molecule from the orientational recurrences, to relate the measured temporal widths to the angular momentum photon state and the coherence of the laser pulse, to obtain information on the ground rotational state, or to measure the effective temperature of an isotropic molecular assembly. The space orientation of a small number of independent molecules can be distinguished from their distinct orientational dependence in the formation of the individual orientational wave packets.

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

    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.

  15. Multiphoton dissociation/ionisation of dimethyl sulphide (CH3SCH3) at 355 and 532 nm

    Rajesh K Vatsa; Chiranjib Majumder; Onnattu D Jayakumar; Shailendra K Kulshreshtha; Jai P Mittal

    2001-04-01

    Multiphoton dissociation/ionization has been studied for CH3SCH3 at 355 and 532 nm using a time-of-flight mass spectrometer. The major ion signals observed at 355 nm are C+, CH$_{3}^{+}$, HCS+, CH2S+, CH3S+ and CH3SCH$_{3}^{+}$. Power dependence studies at 355 nm show a (2 + 1) REMPI process for the formation of parent ion. Peaks at / = 46, 47 and 61 show two-photon laser power dependence whereas / = 15 and 45 peaks show four-photon dependence. However, in 532 nm photoionisation, no parent ion signal is observed. A peak at / = 35 corresponding to SH$_{3}^{+}$ has been observed. SH$_{3}^{+}$ has been suggested to originate from CH3SCH$_{2}^{+}$ via a cyclic transition state. Photoionisation results of CH3SCH3 have been compared with those of CH3SSCH3, at these two wavelengths.

  16. Multi-photon quantum interference in a multi-port integrated photonic device

    Metcalf, Benjamin J; Spring, Justin B; Kundys, Dmytro; Broome, Matthew A; Humphreys, Peter; Jin, Xian-Min; Barbieri, Marco; Kolthammer, W Steven; Gates, James C; Smith, Brian J; Langford, Nathan K; Smith, Peter G R; Walmsley, Ian A

    2012-01-01

    Increasing the complexity of quantum photonic devices is essential for many optical information processing applications to reach a regime beyond what can be classically simulated, and integrated photonics has emerged as a leading platform for achieving this. Here, we demonstrate three-photon quantum operation of an integrated device containing three coupled interferometers, eight spatial modes and many classical and nonclassical interferences. This represents a critical advance over previous complexities and the first on-chip nonclassical interference with more than two photonic inputs. We introduce a new scheme to verify quantum behaviour, using classically characterised device elements and hierarchies of photon correlation functions. We accurately predict the device's quantum behaviour and show operation inconsistent with both classical and bi-separable quantum models. Such methods for verifying multiphoton quantum behaviour are vital for achieving increased circuit complexity. Our experiment paves the way ...

  17. High-order harmonic generation and multi-photon ionization of Na2 in laser fields

    Zhang Yan-Ping; Zhang Feng-Shou; Meng Ke-Lai; Xiao Guo-Qing

    2007-01-01

    In this paper high-order harmonic generation (HHG) spectra and the ionization probabilities of various charge states of small cluster Na2 in the multiphoton regimes are calculated by using time-dependent local density approximation (TDLDA) for one-colour (1064 nm) and two-colour (1064 nm and 532 nm) ultrashort (25 fs) laser pulses. HHG spectra of Na2 have not the large extent of plateaus due to pronounced collective effects of electron dynamics. In addition, the two-colour laser field can result in the breaking of the symmetry and generation of the even order harmonic such as the second order harmonic. The results of ionization probabilities show that a two-colour laser field can increase the ionization probability of higher charge state.

  18. Engineering entangled microwave photon states through multiphoton interactions between two cavity fields and a superconducting qubit

    Zhao, Yan-Jun; Wang, Changqing; Zhu, Xiaobo; Liu, Yu-Xi

    2016-04-01

    It has been shown that there are not only transverse but also longitudinal couplings between microwave fields and a superconducting qubit with broken inversion symmetry of the potential energy. Using multiphoton processes induced by longitudinal coupling fields and frequency matching conditions, we design a universal algorithm to produce arbitrary superpositions of two-mode photon states of microwave fields in two separated transmission line resonators, which are coupled to a superconducting qubit. Based on our algorithm, we analyze the generation of evenly-populated states and NOON states. Compared to other proposals with only single-photon process, we provide an efficient way to produce entangled microwave photon states when the interactions between superconducting qubits and microwave fields are in the strong and ultrastrong regime.

  19. The stepwise multi-photon activation fluorescence guided ablation of melanin

    Lai, Zhenhua; Gu, Zetong; DiMarzio, Charles

    2015-02-01

    Previous research has shown that the stepwise multi-photon activation fluorescence (SMPAF) of melanin, activated and excited by a continuous-wave (CW) mode near infrared (NIR) laser, is a low-cost and reliable method for detecting melanin. We have developed a device utilizing the melanin SMPAF to guide the ablation of melanin with a 975 nm CW laser. This method provides the ability of targeting individual melanin particles with micrometer resolution, and enables localized melanin ablation to be performed without collateral damage. Compared to the traditional selective photothermolysis, which uses pulsed lasers for melanin ablation, this method demonstrates higher precision and lower cost. Therefore, the SMPAF guided selective ablation of melanin is a promising tool of melanin ablation for both medical and cosmetic purposes.

  20. Influence of Heat-radiating on Multi-photon Compton Scattering High-energy Electron

    HAO Dong-shan; WANG Xin-min

    2007-01-01

    Using the model of the inverse Compton scattering between high-energy electrons and heat-radiation photons, the influence of heat-radiating photons on multi-photon Compton scattering high-energy electrons is studied . The results show that the energy loss, power loss, light resistance and light pressure of the high-energy electron formed by heat radiating are all proportional to the temperature T4 of the vacuum cavity of the electron,the Lorentz factor γ2 of the high-energy electrons, the scattering section of the electron and the number of photons acting at the same time with high-energy electrons. A good method for lessening the energy loss of the high-energy electron by using the one-photon Compton scattering between high-energy electrons and heat radiation photons is proposed.