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Sample records for two-photon excitation autofluorescence

  1. Light-induced damage and its diagnosis in two-photon excited autofluorescence imaging of retinal pigment epithelium cells

    Science.gov (United States)

    Chen, Danni; Qu, Junle; Xu, Gaixia; Zhao, Lingling; Niu, Hanben

    2007-05-01

    In this paper, a novel method for the differentiation of the retinal pigment epithelium (RPE) cells after light-induced damage by two-photon excitation is presented. Fresh samples of RPE cells of pig eyes are obtained from local slaughterhouse. Light-induced damage is produced by the output from Ti: sapphire laser which is focused onto the RPE layer. We study the change of the autofluorescence properties of RPE after two-photon excitation with the same wavelength. Preliminary results show that after two-photon excitation, there are two clear changes in the emission spectrum. The first change is the blue-shift of the emission peak. The emission peak of the intact RPE is located at 592nm, and after excitation, it shifts to 540nm. It is supposed that the excitation has led to the increased autofluorescence of flavin whose emission peak is located at 540nm. The second change is the increased intensity of the emission peak, which might be caused by the accelerated aging because the autofluorescence of RPE would increase during aging process. Experimental results indicate that two-photon excitation could not only lead to the damage of the RPE cells in multiphoton RPE imaging, but also provide an evaluation of the light-induced damage.

  2. In vivo spectral imaging of different cell types in the small intestine by two-photon excited autofluorescence

    Science.gov (United States)

    Orzekowsky-Schroeder, Regina; Klinger, Antje; Martensen, Björn; Blessenohl, Maike; Gebert, Andreas; Vogel, Alfred; Hüttmann, Gereon

    2011-11-01

    Spectrally resolved two-photon excited autofluorescence imaging is used to distinguish different cell types and functional areas during dynamic processes in the living gut. Excitation and emission spectra of mucosal tissue and tissue components are correlated to spectra of endogenous chromophores. We show that selective excitation with only two different wavelengths within the tuning range of a Ti:sapphire femtosecond laser system yields excellent discrimination between enterocytes, antigen presenting cells and lysosomes based on the excitation and emission properties of their autofluorescence. The method is employed for time-lapse microscopy over up to 8 h. Changes of the spectral signature with the onset of photodamage are demonstrated, and their origin is discussed.

  3. Age-related structural abnormalities in the human retina-choroid complex revealed by two-photon excited autofluorescence imaging.

    Science.gov (United States)

    Han, Meng; Giese, Guenter; Schmitz-Valckenberg, Steffen; Bindewald-Wittich, Almut; Holz, Frank G; Yu, Jiayi; Bille, Josef F; Niemz, Markolf H

    2007-01-01

    The intensive metabolism of photoreceptors is delicately maintained by the retinal pigment epithelium (RPE) and the choroid. Dysfunction of either the RPE or choroid may lead to severe damage to the retina. Two-photon excited autofluorescence (TPEF) from endogenous fluorophores in the human retina provides a novel opportunity to reveal age-related structural abnormalities in the retina-choroid complex prior to apparent pathological manifestations of age-related retinal diseases. In the photoreceptor layer, the regularity of the macular photoreceptor mosaic is preserved during aging. In the RPE, enlarged lipofuscin granules demonstrate significantly blue-shifted autofluorescence, which coincides with the depletion of melanin pigments. Prominent fibrillar structures in elderly Bruch's membrane and choriocapillaries represent choroidal structure and permeability alterations. Requiring neither slicing nor labeling, TPEF imaging is an elegant and highly efficient tool to delineate the thick, fragile, and opaque retina-choroid complex, and may provide clues to the trigger events of age-related macular degeneration.

  4. Two-photon autofluorescence spectroscopy of oral mucosa tissue

    Science.gov (United States)

    Edward, Kert; Shilagard, Tuya; Qiu, Suimin; Vargas, Gracie

    2011-03-01

    The survival rate for individuals diagnosed with oral cancer is correlated with the stage of detection. Thus the development of novel techniques for the earliest possible detection of malignancies is of critical importance. Single photon (1P) autofluorescence spectroscopy has proven to be a powerful diagnostic tool in this regard, but 2P (two photon) spectroscopy remains essentially unexplored. In this investigation, a spectroscopic system was incorporated into a custom-built 2P laser scanning microscope. Oral cancer was induced in the buccal pouch of Syrian Golden hamsters by tri-weekly topical application of 9,10-dimethyl-1,2-benzanthracene (DMBA).Three separated sites where investigated in each hamster at four excitation wavelengths from 780 nm to 890 nm. A Total of 8 hamsters were investigated (4 normal and 4 DMBA treated). All investigated sites were imaged via 2p imaging, marked for biopsy, processed for histology and H&E staining, and graded by a pathologist. The in vivo emission spectrum for normal, mild/high grade dysplasia and squamous cell carcinoma is presented. It is shown that the hamsters with various stages of dysplasia are characterized by spectral differences as a function of depth and excitation wavelength, compared to normal hamsters.

  5. Quantitative imaging of fibrotic and morphological changes in liver of non-alcoholic steatohepatitis (NASH) model mice by second harmonic generation (SHG) and auto-fluorescence (AF) imaging using two-photon excitation microscopy (TPEM).

    Science.gov (United States)

    Yamamoto, Shin; Oshima, Yusuke; Saitou, Takashi; Watanabe, Takao; Miyake, Teruki; Yoshida, Osamu; Tokumoto, Yoshio; Abe, Masanori; Matsuura, Bunzo; Hiasa, Yoichi; Imamura, Takeshi

    2016-12-01

    Non-alcoholic steatohepatitis (NASH) is a common liver disorder caused by fatty liver. Because NASH is associated with fibrotic and morphological changes in liver tissue, a direct imaging technique is required for accurate staging of liver tissue. For this purpose, in this study we took advantage of two label-free optical imaging techniques, second harmonic generation (SHG) and auto-fluorescence (AF), using two-photon excitation microscopy (TPEM). Three-dimensional ex vivo imaging of tissues from NASH model mice, followed by image processing, revealed that SHG and AF are sufficient to quantitatively characterize the hepatic capsule at an early stage and parenchymal morphologies associated with liver disease progression, respectively.

  6. Two-photon autofluorescence lifetime and SHG imaging of healthy and diseased human corneas

    Science.gov (United States)

    Batista, Ana; Breunig, Hans Georg; Uchugonova, Aisada; Seitz, Berthold; Morgado, António Miguel; König, Karsten

    2015-03-01

    Corneal function can be drastically affected by several degenerations and dystrophies, leading to blindness. Early diagnosis of corneal disease is of major importance and it may be accomplished by monitoring changes of the metabolic state and structural organization, the first detectable pathological signs, by two-photon excitation autofluorescence lifetime and second-harmonic generation imaging. In this study, we propose to use these imaging techniques to differentiate between healthy and pathological corneas. Images were acquired using a laser-scanning microscope with a broadband sub-15 femtosecond near-infrared pulsed laser and a 16-channel photomultiplier tube detector for signal collection. This setup allows the simultaneous excitation of metabolic co-factors and to identify them based on their fluorescence spectra. We were able to discriminate between healthy and pathological corneas using two-photon excitation autofluorescence lifetime and second-harmonic generation imaging from corneal epithelium and stroma. Furthermore, differences between different pathologies were observed. Alterations in the metabolic state of corneal epithelial cells were observed using the autofluorescence lifetime of the metabolic co-factors. In the corneal stroma, we observed not only alterations in the collagen fibril structural organization but also alterations in the autofluorescence lifetime. Further tests are required as the number of pathological samples must be increased. In the future, we intend to establish a correlation between the metabolic and structural changes and the disease stage. This can be a step forward in achieving early diagnosis.

  7. Effects of Ox-LDL on Macrophages NAD(P)H Autofluorescence Changes by Two-photon Microscopy

    CERN Document Server

    Lin, Ching-Ting; Lee, Szu-Yuan; Lu, Long-Sheng; Wu, Chau-Chung; Dong, Chen-Yuan; Lin, Chii-Wann

    2007-01-01

    Ox-LDL uptakes by macrophage play a critical role in the happening of atherosclerosis. Because of its low damage on observed cells and better signal-to- background ratio, two-photon excitation fluorescence microscopy is used to observe NAD(P)H autofluorescence of macrophage under difference cultured conditions- bare cover glass, coated with fibronectin or poly-D-lysine. The results show that the optimal condition is fibronectin coated surface, on which, macrophages profile can be clearly identified on NAD(P)H autofluorescence images collected by two-photon microscopy. Moreover, different morphology and intensities of autofluorescence under different conditions were observed as well. In the future, effects of ox-LDL on macrophages will be investigated by purposed system to research etiology of atherosclerosis.

  8. Two-photon excited spectroscopies of ex vivo human skin endogenous species irradiated by femtosecond laser pulses

    Institute of Scientific and Technical Information of China (English)

    Jianxin Chen; Shuangmu Zhuo; Tianshu Luo; Jingjun Zhao

    2006-01-01

    Two-photon excited spectroscopies from ex vivo human skin are investigated by using a femtosecond laser and a confocal microscope (Zeiss LSM 510 META). In the dermis, collagen is responsible for second harmonic generation (SHG); elastin, nicotinamide adenine dinucleotide (NADH), melanin and porphyrin are the primary endogenous sources of two-photon excited autofluorescence. In the epidermis, keratin,NADH, melanin and porphyrins contribute to autofluorescence signals. The results also show that the SHG spectra have the ability to shift with the excitation wavelength and the autofluorescence spectra display a red shift of the spectral peaks when increasing the excitation wavelength. These results may have practical implications for diagnosis of skin diseases.

  9. Two-photon excited spectroscopies of ex vivo human skin endogenous species irradiated by femtosecond laser pulses

    Science.gov (United States)

    Chen, Jianxin; Zhuo, Shuangmu; Luo, Tianshu; Zhao, Jingjun

    2006-10-01

    Two-photon excited spectroscopies from ex vivo human skin are investigated by using a femtosecond laser and a confocal microscope (Zeiss LSM 510 META). In the dermis, collagen is responsible for second harmonic generation (SHG); elastin, nicotinamide adenine dinucleotide (NADH), melanin and porphyrin are the primary endogenous sources of two-photon excited autofluorescence. In the epidermis, keratin, NADH, melanin and porphyrins contribute to autofluorescence signals. The results also show that the SHG spectra have the ability to shift with the excitation wavelength and the autofluorescence spectra display a red shift of the spectral peaks when increasing the excitation wavelength. These results may have practical implications for diagnosis of skin diseases.

  10. Two-photon-excited autofluorescence and second-harmonic generation microscopy for the visualization of penetration of TiO2 and ZnO nanoparticles into human tooth tissue ex vivo

    Science.gov (United States)

    Trunina, Natalia A.; Popov, Alexey P.; Lademann, Jürgen; Tuchin, Valery V.; Myllylä, Risto; Darvin, Maxim E.

    2012-06-01

    Penetration of nanoparticles into tooth tissues is of significant interest in solving problems related to reduction of tooth sensitivity, enamel strengthening and restoration and cosmetic bleaching. In this work we demonstrate two-photonexcited autofluorescence and second-harmonic generation microscopy for visualization of penetration of TiO2 and ZnO nanoparticles into tooth tissues.

  11. Simultaneous two-photon excitation of photodynamic therapy agents

    Energy Technology Data Exchange (ETDEWEB)

    Wachter, E.A.; Fisher, W.G. [Oak Ridge National Lab., TN (United States)]|[Photogen, Inc., Knoxville, TN (United States); Partridge, W.P. [Oak Ridge National Lab., TN (United States); Dees, H.C. [Photogen, Inc., Knoxville, TN (United States); Petersen, M.G. [Univ. of Tennessee, Knoxville, TN (United States). College of Veterinary Medicine

    1998-01-01

    The spectroscopic and photochemical properties of several photosensitive compounds are compared using conventional single-photon excitation (SPE) and simultaneous two-photon excitation (TPE). TPE is achieved using a mode-locked titanium:sapphire laser, the near infrared output of which allows direct promotion of non-resonant TPE. Excitation spectra and excited state properties of both type 1 and type 2 photodynamic therapy (PDT) agents are examined.

  12. Two-photon excited ultraviolet photoluminescence of zinc oxide nanorods.

    Science.gov (United States)

    Zhu, Guangping; Xu, Chunxiang; Zhu, Jing; Lu, Changgui; Cui, Yiping; Sun, Xiaowei

    2008-11-01

    High density zinc oxide nanorods with uniform size were synthesized on (100) silicon substrate by vapor-phase transport method. The scanning electron microscopy images reveal that the nanorods have an average diameter of about 400 nm. The X-ray diffraction pattern demonstrates the wurtzite crystalline structure of the ZnO nanorods growing along [0001] direction. The single-photon excited photoluminescence presents a strong ultraviolet emission band at 394 nm and a weak visible emission band at 600 nm. When the ZnO nanorods were respectively pumped by various wavelength lasers from 520 nm to 700 nm, two-photon excited ultraviolet photoluminescence was observed. The dependence of the two-photon excited photoluminescence intensity on the excitation wavelength and power was investigated in detail.

  13. Diagnosis of basal cell carcinoma by two photon excited fluorescence combined with lifetime imaging

    Science.gov (United States)

    Fan, Shunping; Peng, Xiao; Liu, Lixin; Liu, Shaoxiong; Lu, Yuan; Qu, Junle

    2014-02-01

    Basal cell carcinoma (BCC) is the most common type of human skin cancer. The traditional diagnostic procedure of BCC is histological examination with haematoxylin and eosin staining of the tissue biopsy. In order to reduce complexity of the diagnosis procedure, a number of noninvasive optical methods have been applied in skin examination, for example, multiphoton tomography (MPT) and fluorescence lifetime imaging microscopy (FLIM). In this study, we explored two-photon optical tomography of human skin specimens using two-photon excited autofluorescence imaging and FLIM. There are a number of naturally endogenous fluorophores in skin sample, such as keratin, melanin, collagen, elastin, flavin and porphyrin. Confocal microscopy was used to obtain structures of the sample. Properties of epidermic and cancer cells were characterized by fluorescence emission spectra, as well as fluorescence lifetime imaging. Our results show that two-photon autofluorescence lifetime imaging can provide accurate optical biopsies with subcellular resolution and is potentially a quantitative optical diagnostic method in skin cancer diagnosis.

  14. Simultaneous two-photon excitation of photodynamic therapy agents

    Science.gov (United States)

    Wachter, Eric A.; Partridge, W. P., Jr.; Fisher, Walter G.; Dees, Craig; Petersen, Mark G.

    1998-07-01

    The spectroscopic and photochemical properties of several photosensitive compounds are compared using conventional single-photon excitation (SPE) and simultaneous two-photon excitation (TPE). TPE is achieved using a mode-locked titanium:sapphire laser, the near infrared output of which allows direct promotion of non-resonant TPE. Excitation spectra and excited state properties of both type I and type II photodynamic therapy (PDT) agents are examined. In general, while SPE and TPE selection rules may be somewhat different, the excited state photochemical properties are equivalent for both modes of excitation. In vitro promotion of a two-photon photodynamic effect is demonstrated using bacterial and human breast cancer models. These results suggest that use of TPE may be beneficial for PDT, since the technique allows replacement of visible or ultraviolet excitation with non- damaging near infrared light. Further, a comparison of possible excitation sources for TPE indicates that the titanium:sapphire laser is exceptionally well suited for non- linear excitation of PDT agents in biological systems due to its extremely short pulse width and high repetition rate; these features combine to effect efficient PDT activation with minimal potential for non-specific biological damage.

  15. Maximum imaging depth of two-photon autofluorescence microscopy in epithelial tissues.

    Science.gov (United States)

    Durr, Nicholas J; Weisspfennig, Christian T; Holfeld, Benjamin A; Ben-Yakar, Adela

    2011-02-01

    Endogenous fluorescence provides morphological, spectral, and lifetime contrast that can indicate disease states in tissues. Previous studies have demonstrated that two-photon autofluorescence microscopy (2PAM) can be used for noninvasive, three-dimensional imaging of epithelial tissues down to approximately 150 μm beneath the skin surface. We report ex-vivo 2PAM images of epithelial tissue from a human tongue biopsy down to 370 μm below the surface. At greater than 320 μm deep, the fluorescence generated outside the focal volume degrades the image contrast to below one. We demonstrate that these imaging depths can be reached with 160 mW of laser power (2-nJ per pulse) from a conventional 80-MHz repetition rate ultrafast laser oscillator. To better understand the maximum imaging depths that we can achieve in epithelial tissues, we studied image contrast as a function of depth in tissue phantoms with a range of relevant optical properties. The phantom data agree well with the estimated contrast decays from time-resolved Monte Carlo simulations and show maximum imaging depths similar to that found in human biopsy results. This work demonstrates that the low staining inhomogeneity (∼ 20) and large scattering coefficient (∼ 10 mm(-1)) associated with conventional 2PAM limit the maximum imaging depth to 3 to 5 mean free scattering lengths deep in epithelial tissue.

  16. Four-dimensional multi-site two-photon excitation

    CERN Document Server

    Daria, Vincent Ricardo; Bowman, Richard; Redman, Stephen; Bachor, Hans-A

    2009-01-01

    We report the first demonstration of dynamic and arbitrary multi-site two-photon excitation in three-dimensional (3D) space using the holographic projection method. Rapid temporal response (fourth dimension) is achieved through high-speed non-iterative and non-optimized calculation of the hologram using a video graphics accelerator board. We verify that the projected asymmetric spot configurations have sufficient spatiotemporal photon density for localized two-photon excitation. This system is a significant advance and ready for applications such as time-resolved 3D photolysis of complex biological cell and neuronal networks, 3D microscopy, non-linear micro-fabrication and volume holographic optical storage.

  17. Two-photon microscopy using fiber-based nanosecond excitation.

    Science.gov (United States)

    Karpf, Sebastian; Eibl, Matthias; Sauer, Benjamin; Reinholz, Fred; Hüttmann, Gereon; Huber, Robert

    2016-07-01

    Two-photon excitation fluorescence (TPEF) microscopy is a powerful technique for sensitive tissue imaging at depths of up to 1000 micrometers. However, due to the shallow penetration, for in vivo imaging of internal organs in patients beam delivery by an endoscope is crucial. Until today, this is hindered by linear and non-linear pulse broadening of the femtosecond pulses in the optical fibers of the endoscopes. Here we present an endoscope-ready, fiber-based TPEF microscope, using nanosecond pulses at low repetition rates instead of femtosecond pulses. These nanosecond pulses lack most of the problems connected with femtosecond pulses but are equally suited for TPEF imaging. We derive and demonstrate that at given cw-power the TPEF signal only depends on the duty cycle of the laser source. Due to the higher pulse energy at the same peak power we can also demonstrate single shot two-photon fluorescence lifetime measurements.

  18. Enhanced multi-spectral imaging of live breast cancer cells using immunotargeted gold nanoshells and two-photon excitation microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Bickford, Lissett; Sun Jiantang; Fu, Kun; Lewinski, Nastassja; Nammalvar, Vengadesan; Chang, Joseph; Drezek, Rebekah [Department of Bioengineering, Rice University, Houston, TX 77005 (United States)], E-mail: drezek@rice.edu

    2008-08-06

    We demonstrate the capability of using immunotargeted gold nanoshells as contrast agents for in vitro two-photon microscopy. The two-photon luminescence properties of different-sized gold nanoshells are first validated using near-infrared excitation at 780 nm. The utility of two-photon microscopy as a tool for imaging live HER2-overexpressing breast cancer cells labeled with anti-HER2-conjugated nanoshells is then explored and imaging results are compared to normal breast cells. Five different imaging channels are simultaneously examined within the emission wavelength range of 451-644 nm. Our results indicate that under near-infrared excitation, superior contrast of SK-BR-3 cancer cells labeled with immunotargeted nanoshells occurs at an emission wavelength ranging from 590 to 644 nm. Luminescence from labeled normal breast cells and autofluorescence from unlabeled cancer and normal cells remain imperceptible under the same conditions.

  19. Enhanced multi-spectral imaging of live breast cancer cells using immunotargeted gold nanoshells and two-photon excitation microscopy

    Science.gov (United States)

    Bickford, Lissett; Sun, Jiantang; Fu, Kun; Lewinski, Nastassja; Nammalvar, Vengadesan; Chang, Joseph; Drezek, Rebekah

    2008-08-01

    We demonstrate the capability of using immunotargeted gold nanoshells as contrast agents for in vitro two-photon microscopy. The two-photon luminescence properties of different-sized gold nanoshells are first validated using near-infrared excitation at 780 nm. The utility of two-photon microscopy as a tool for imaging live HER2-overexpressing breast cancer cells labeled with anti-HER2-conjugated nanoshells is then explored and imaging results are compared to normal breast cells. Five different imaging channels are simultaneously examined within the emission wavelength range of 451-644 nm. Our results indicate that under near-infrared excitation, superior contrast of SK-BR-3 cancer cells labeled with immunotargeted nanoshells occurs at an emission wavelength ranging from 590 to 644 nm. Luminescence from labeled normal breast cells and autofluorescence from unlabeled cancer and normal cells remain imperceptible under the same conditions.

  20. Two-photon excitation photodynamic therapy with Photofrin

    Science.gov (United States)

    Karotki, Aliaksandr; Khurana, Mamta; Lepock, James R.; Wilson, Brian C.

    2005-09-01

    Photodynamic therapy (PDT) based on simultaneous two-photon (2-γ) excitation has a potential advantage of highly targeted treatment by means of nonlinear localized photosensitizer excitation. One of the possible applications of 2-γ PDT is a treatment of exodus age-related macular degeneration where highly targeted excitation of photosensitizer in neovasculature is vital for reducing collateral damage to healthy surrounding tissue. To investigate effect of 2-γ PDT Photofrin was used as an archetypal photosensitizer. First, 2-γ absorption properties of Photofrin in the 750 - 900 nm excitation wavelength range were investigated. It was shown that above 800 nm 2-γ interaction was dominant mode of excitation. The 2-γ cross section of Photofrin was rather small and varied between 5 and 10 GM (1 GM = 10-50 cm4s/photon) in this wavelength range. Next, endothelial cells treated with Photofrin were used to model initial effect of 2-γ PDT on neovasculature. Ultrashort laser pulses provided by mode-locked Ti:sapphire laser (pulse duration at the sample 300 fs, repetition rate 90 MHz, mean laser power 10 mW, excitation wavelength 850 nm) were used for the excitation of the photosensitizer. Before 2-γ excitation of the Photofrin cells formed a single continuous sheet at the bottom of the well. The tightly focused laser light was scanned repeatedly over the cell layer. After irradiation the cell layer of the control cells stayed intact while cells treated with photofrin became clearly disrupted. The light doses required were high (6300 Jcm(-2) for ~ 50% killing), but 2-γ cytotoxicity was unequivocally demonstrated.

  1. Two-photon excited photoconversion of cyanine-based dyes

    Science.gov (United States)

    Kwok, Sheldon J. J.; Choi, Myunghwan; Bhayana, Brijesh; Zhang, Xueli; Ran, Chongzhao; Yun, Seok-Hyun

    2016-03-01

    The advent of phototransformable fluorescent proteins has led to significant advances in optical imaging, including the unambiguous tracking of cells over large spatiotemporal scales. However, these proteins typically require activating light in the UV-blue spectrum, which limits their in vivo applicability due to poor light penetration and associated phototoxicity on cells and tissue. We report that cyanine-based, organic dyes can be efficiently photoconverted by nonlinear excitation at the near infrared (NIR) window. Photoconversion likely involves singlet-oxygen mediated photochemical cleavage, yielding blue-shifted fluorescent products. Using SYTO62, a biocompatible and cell-permeable dye, we demonstrate photoconversion in a variety of cell lines, including depth-resolved labeling of cells in 3D culture. Two-photon photoconversion of cyanine-based dyes offer several advantages over existing photoconvertible proteins, including use of minimally toxic NIR light, labeling without need for genetic intervention, rapid kinetics, remote subsurface targeting, and long persistence of photoconverted signal. These findings are expected to be useful for applications involving rapid labeling of cells deep in tissue.

  2. Mapping of hemoglobin in erythrocytes and erythrocyte ghosts using two photon excitation fluorescence microscopy

    Science.gov (United States)

    Bukara, Katarina; Jovanić, Svetlana; Drvenica, Ivana T.; Stančić, Ana; Ilić, Vesna; Rabasović, Mihailo D.; Pantelić, Dejan; Jelenković, Branislav; Bugarski, Branko; Krmpot, Aleksandar J.

    2017-02-01

    The present study describes utilization of two photon excitation fluorescence (2PE) microscopy for visualization of the hemoglobin in human and porcine erythrocytes and their empty membranes (i.e., ghosts). High-quality, label- and fixation-free visualization of hemoglobin was achieved at excitation wavelength 730 nm by detecting visible autofluorescence. Localization in the suspension and spatial distribution (i.e., mapping) of residual hemoglobin in erythrocyte ghosts has been resolved by 2PE. Prior to the 2PE mapping, the presence of residual hemoglobin in the bulk suspension of erythrocyte ghosts was confirmed by cyanmethemoglobin assay. 2PE analysis revealed that the distribution of hemoglobin in intact erythrocytes follows the cells' shape. Two types of erythrocytes, human and porcine, characterized with discocyte and echinocyte morphology, respectively, showed significant differences in hemoglobin distribution. The 2PE images have revealed that despite an extensive washing out procedure after gradual hypotonic hemolysis, a certain amount of hemoglobin localized on the intracellular side always remains bound to the membrane and cannot be eliminated. The obtained results open the possibility to use 2PE microscopy to examine hemoglobin distribution in erythrocytes and estimate the purity level of erythrocyte ghosts in biotechnological processes.

  3. Two-photon autofluorescence/FLIM/SHG endoscopy to study the oral cavity and wound healing in humans (Conference Presentation)

    Science.gov (United States)

    König, Karsten

    2016-03-01

    Monitoring the oral cavity noninvasively with superior 3D resolution is realized by clinical multiphoton tomography and high NA two-photon endoscopy without the need of additional contrast agents. The technology behind this investigation is based on nonlinear optical contrast of the multiphoton tomograph MPTflex®. Furthermore, the miniaturized GRIN endoscope was used to realize more accessibility for more demanding wound conditions in skin. The MPTflex® distinguishes autofluorescence (AF) signals from second harmonic generation (SHG) signals simultaneously. Fluorescence lifetime imaging (FLIM) based on time correlated single photon counting (TCSPC) technology offers additional information on the functional level of the intratissue fluorophores, their binding status, and the contribution of SHG signals in chronic wounds.

  4. Tracking of mercury ions in living cells with a fluorescent chemodosimeter under single- or two-photon excitation

    Energy Technology Data Exchange (ETDEWEB)

    Lu Zhoujun [State Key Lab for Advanced Photonic Materials and Devices, Department of Optical Science and Engineering, Fudan University, Shanghai 200433 (China); Wang Peinan [State Key Lab for Advanced Photonic Materials and Devices, Department of Optical Science and Engineering, Fudan University, Shanghai 200433 (China)], E-mail: pnwang@fudan.edu.cn; Zhang Yu [State Key Lab for Advanced Photonic Materials and Devices, Department of Optical Science and Engineering, Fudan University, Shanghai 200433 (China); Chen Jiyao; Zhen Shen [Department of Physics, Fudan University, Shanghai 200433 (China); Leng Bing; Tian He [Labs for Advanced Materials and Institute of Fine Chemicals, East China University of Science and Technology, Shanghai 200237 (China)

    2007-08-10

    Tracking of Hg{sup 2+} in solutions as well as in living cells was conducted with a fluorescent chemodosimeter by measuring the spectral shift of its fluorescence under single- or two-photon excitation. The spectral hypsochromic shifts of this chemodosimeter when reacting with Hg{sup 2+} were found to be about 50 nm in acetonitrile/water solutions and 32 nm in Euglena gracilis 277 living cells. This chemodosimeter shows high sensitivity and selectivity, and is not influenced by the pH values. It can signal Hg{sup 2+} in solutions down to the ppb range under either single-photon excitation (SPE) at 405 nm or two-photon excitation (TPE) at 800 nm. However, with low cellular chemodosimeter concentrations, the SPE spectra were disturbed by the auto-fluorescence from the native fluorophore in the cell, while the TPE spectra were still of high quality since the two-photon absorption cross section of this chemodosimeter is much larger than that of the native fluorophores in the cell.

  5. LANTHANIDE ENHANCE LUMINESCENCE (LEL) WITH ONE AND TWO PHOTON EXCITATION OF QUANTUM DYES LANTHANIDE (III) - MACROCYCLES

    Science.gov (United States)

    Title: Lanthanide Enhance Luminescence (LEL) with one and two photon excitation of Quantum Dyes? Lanthanide(III)-Macrocycles Principal Author:Robert C. Leif, Newport InstrumentsSecondary Authors:Margie C. Becker, Phoenix Flow Systems Al Bromm, Virginia Commonw...

  6. LANTHANIDE ENHANCE LUMINESCENCE (LEL) WITH ONE AND TWO PHOTON EXCITATION OF QUANTUM DYES LANTHANIDE (III) - MACROCYCLES

    Science.gov (United States)

    Title: Lanthanide Enhance Luminescence (LEL) with one and two photon excitation of Quantum Dyes? Lanthanide(III)-Macrocycles Principal Author:Robert C. Leif, Newport InstrumentsSecondary Authors:Margie C. Becker, Phoenix Flow Systems Al Bromm, Virginia Commonw...

  7. Two-photon vibrational excitation of air by long-wave infrared laser pulses

    CERN Document Server

    Palastro, J P; Johnson, L A; Hafizi, B; Wahlstrand, J K; Milchberg, H M

    2016-01-01

    Ultrashort long-wave infrared (LWIR) laser pulses can resonantly excite vibrations in N2 and O2 through a two-photon transition. The absorptive, vibrational component of the ultrafast optical nonlinearity grows in time, starting smaller than, but quickly surpassing, the electronic, rotational, and vibrational refractive components. The growth of the vibrational component results in a novel mechanism of 3rd harmonic generation, providing an additional two-photon excitation channel, fundamental + 3rd harmonic. The original and emergent two-photon excitations drive the resonance exactly out of phase, causing spatial decay of the absorptive, vibrational nonlinearity. This nearly eliminates two-photon vibrational absorption. Here we present simulations and analytical calculations demonstrating how these processes modify the ultrafast optical nonlinearity in air. The results reveal nonlinear optical phenomena unique to the LWIR regime of ultrashort pulse propagation in atmosphere.

  8. Arduino Due based tool to facilitate in vivo two-photon excitation microscopy.

    Science.gov (United States)

    Artoni, Pietro; Landi, Silvia; Sato, Sebastian Sulis; Luin, Stefano; Ratto, Gian Michele

    2016-04-01

    Two-photon excitation spectroscopy is a powerful technique for the characterization of the optical properties of genetically encoded and synthetic fluorescent molecules. Excitation spectroscopy requires tuning the wavelength of the Ti:sapphire laser while carefully monitoring the delivered power. To assist laser tuning and the control of delivered power, we developed an Arduino Due based tool for the automatic acquisition of high quality spectra. This tool is portable, fast, affordable and precise. It allowed studying the impact of scattering and of blood absorption on two-photon excitation light. In this way, we determined the wavelength-dependent deformation of excitation spectra occurring in deep tissues in vivo.

  9. Two-Photon Excited Fluorescence from Biological Aerosol Particles

    Science.gov (United States)

    2010-09-29

    previously observed from serotonin (5-HT) and its precursor hyrdroxytryptophan (5- HTP ) using multi-photon excitation [17-19]. Visible emission from...Sivaprakasam, A. Huston, H.B. Lin, J.D. Eversole, P. Falkenstein and A. Schultz, “Field test results and ambient aerosol measurements using dual

  10. Autofluorescence excitation-emission matrices for diagnosis of colonic cancer

    Institute of Scientific and Technical Information of China (English)

    Bu-Hong Li; Shu-Sen Xie

    2005-01-01

    AIM: To investigate the autofluorescence spectroscopic differences in normal and adenomatous colonic tissues and to determine the optimal excitation wavelengths for subsequent study and clinical application.METHODS: Normal and adenomatous colonic tissues were obtained from patients during surgery. A FL/FS920combined TCSPC spectrofluorimeter and a lifetime spectrometer system were used for fluorescence measurement.Fluorescence excitation wavelengths varying from 260 to 540 nm were used to induce the autofluorescence spectra,and the corresponding emission spectra were recorded from a range starting 20 nm above the excitation wavelength and extending to 800 nm. Emission spectra were assembled into a three-dimensional fluorescence spectroscopy and an excitation-emission matrix (EEM) to exploit endogenous fluorophores and diagnostic information. Then emission spectra of normal and adenomatous colonic tissues at certain excitation wavelengths were compared to determine the optimal excitation wavelengths for diagnosis of colonic cancer.RESULTS: When compared to normal tissues, low NAD (P)H and FAD, but high amino acids and endogenous phorphyrins of protoporphyrin Ⅸ characterized the highgrade malignant colonic tissues. The optimal excitation wavelengths for diagnosis of colonic cancer were about 340, 380, 460, and 540 nm.CONCLUSION: Significant differences in autofluorescence peaks and its intensities can be observed in normal and adenomatous colonic tissues. Autofluorescence EEMs are able to identify colonic tissues.

  11. Cyanines as new fluorescent probes for DNA detection and two-photon excited bioimaging.

    Science.gov (United States)

    Feng, Xin Jiang; Wu, Po Lam; Bolze, Frédéric; Leung, Heidi W C; Li, King Fai; Mak, Nai Ki; Kwong, Daniel W J; Nicoud, Jean-François; Cheah, Kok Wai; Wong, Man Shing

    2010-05-21

    A series of cyanine fluorophores based on fused aromatics as an electron donor for DNA sensing and two-photon bioimaging were synthesized, among which the carbazole-based biscyanine exhibits high sensitivity and efficiency as a fluorescent light-up probe for dsDNA, which shows selective binding toward the AT-rich regions. The synergetic effect of the bischromophoric skeleton gives a several-fold enhancement in a two-photon absorption cross-section as well as a 25- to 100-fold enhancement in two-photon excited fluorescence upon dsDNA binding.

  12. Two-photon-excited fluorescence spectroscopy of atomic fluorine at 170 nm

    Science.gov (United States)

    Herring, G. C.; Dyer, Mark J.; Jusinski, Leonard E.; Bischel, William K.

    1988-01-01

    Two-photon-excited fluorescence spectroscopy of atomic fluorine is reported. A doubled dye laser at 286-nm is Raman shifted in H2 to 170 nm (sixth anti-Stokes order) to excite ground-state 2P(0)J fluorine atoms to the 2D(0)J level. The fluorine atoms are detected by one of two methods: observing the fluorescence decay to the 2PJ level or observing F(+) production through the absorption of an additional photon by the excited atoms. Relative two-photon absorption cross sections to and the radiative lifetimes of the 2D(0)J states are measured.

  13. Decay and coherence of two-photon excited yellow orthoexcitons in Cu2O

    NARCIS (Netherlands)

    Karpinska, Katarzyna; Mostovoy, M; van der Vegte, MA; Revcolevschi, A; van Loosdrecht, PHM

    2005-01-01

    Photoluminescence excitation spectroscopy has revealed a highly efficient two-photon excitation method to produce a cold, uniformly distributed high density excitonic gas in bulk cuprous oxide. A study of the time evolution of the density, temperature, and chemical potential of the exciton gas shows

  14. Applying two-photon excitation fluorescence lifetime imaging microscopy to study photosynthesis in plant leaves

    NARCIS (Netherlands)

    Broess, K.; Borst, J.W.; Amerongen, van H.

    2009-01-01

    This study investigates to which extent two-photon excitation (TPE) fluorescence lifetime imaging microscopy can be applied to study picosecond fluorescence kinetics of individual chloroplasts in leaves. Using femtosecond 860 nm excitation pulses, fluorescence lifetimes can be measured in leaves of

  15. Two-photon excitation with pico-second fluorescence lifetime imaging to detect nuclear association of flavanols

    Energy Technology Data Exchange (ETDEWEB)

    Mueller-Harvey, Irene, E-mail: i.mueller-harvey@reading.ac.uk [Chemistry and Biochemistry Laboratory, Food Production and Quality Research Division, School of Agriculture, Policy and Development, University of Reading, P O Box 236, Reading RG6 6AT (United Kingdom); Feucht, Walter, E-mail: walter.feucht@gmail.com [Department of Plant Sciences, Technical University of Munich (TUM), Wissenschaftszentrum Weihenstephan (WZW), D-85354 Freising (Germany); Polster, Juergen, E-mail: j.polster@wzw.tum.de [Department of Physical Biochemistry, Technical University of Munich (TUM), Wissenschaftszentrum Weihenstephan (WZW), D-85354 Freising (Germany); Trnkova, Lucie, E-mail: lucie.trnkova@uhk.cz [University of Hradec Kralove, Faculty of Science, Department of Chemistry, Rokitanskeho 62, 50003 Hradec Kralove (Czech Republic); Burgos, Pierre, E-mail: pierre.burgos@stfc.ac.uk [Central Laser Facility, Research Complex at Harwell, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell-Oxford, Didcot, Oxfordshire, OX11 0QX (United Kingdom); Parker, Anthony W., E-mail: tony.parker@stfc.ac.uk [Central Laser Facility, Research Complex at Harwell, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell-Oxford, Didcot, Oxfordshire, OX11 0QX (United Kingdom); Botchway, Stanley W., E-mail: stan.botchway@stfc.ac.uk [Central Laser Facility, Research Complex at Harwell, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell-Oxford, Didcot, Oxfordshire, OX11 0QX (United Kingdom)

    2012-03-16

    Highlights: Black-Right-Pointing-Pointer This fluorescence lifetime imaging microscopy (FLIM) technique for flavanols overcomes autofluorescence interference in cells. Black-Right-Pointing-Pointer Plant flavanols differed in their lifetimes. Black-Right-Pointing-Pointer Dissolved and bound flavanols revealed contrasting lifetime changes. Black-Right-Pointing-Pointer This technique will allow studying of flavanol trafficking in live cells. - Abstract: Two-photon excitation enabled for the first time the observation and measurement of excited state fluorescence lifetimes from three flavanols in solution, which were {approx}1.0 ns for catechin and epicatechin, but <45 ps for epigallocatechin gallate (EGCG). The shorter lifetime for EGCG is in line with a lower fluorescence quantum yield of 0.003 compared to catechin (0.015) and epicatechin (0.018). In vivo experiments with onion cells demonstrated that tryptophan and quercetin, which tend to be major contributors of background fluorescence in plant cells, have sufficiently low cross sections for two-photon excitation at 630 nm and therefore do not interfere with detection of externally added or endogenous flavanols in Allium cepa or Taxus baccata cells. Applying two-photon excitation to flavanols enabled 3-D fluorescence lifetime imaging microscopy and showed that added EGCG penetrated the whole nucleus of onion cells. Interestingly, EGCG and catechin showed different lifetime behaviour when bound to the nucleus: EGCG lifetime increased from <45 to 200 ps, whilst catechin lifetime decreased from 1.0 ns to 500 ps. Semi-quantitative measurements revealed that the relative ratios of EGCG concentrations in nucleoli associated vesicles: nucleus: cytoplasm were ca. 100:10:1. Solution experiments with catechin, epicatechin and histone proteins provided preliminary evidence, via the appearance of a second lifetime ({tau}{sub 2} = 1.9-3.1 ns), that both flavanols may be interacting with histone proteins. We conclude that there

  16. Investigation of two-photon absorption induced excited state absorption in a fluorenyl-based chromophore.

    Science.gov (United States)

    Li, Changwei; Yang, Kun; Feng, Yan; Su, Xinyan; Yang, Junyi; Jin, Xiao; Shui, Min; Wang, Yuxiao; Zhang, Xueru; Song, Yinglin; Xu, Hongyao

    2009-12-03

    Two-photon absorption induced excited state absorption in the solution of a new fluorenyl-based chromophore is investigated by a time-resolved pump-probe technique using femtosecond pulses. With the help of an additional femtosecond open-aperture Z-scan technique, numerical simulations based on a three-energy level model are used to interpret the experimental results, and we determine the nonlinear optical parameters of this new chromophore uniquely. Large two-photon absorption cross section and excited state absorption cross section for singlet excited state are obtained, indicating a good candidate for optical limiting devices. Moreover, the influence of two-beam coupling induced energy transfer in neat N,N'-dimethylformamide solvent is also considered, although this effect is strongly restrained by the instantaneous two-photon absorption.

  17. Selective two-photon excitation of a vibronic state by correlated photons.

    Science.gov (United States)

    Oka, Hisaki

    2011-03-28

    We theoretically investigate the two-photon excitation of a molecular vibronic state by correlated photons with energy anticorrelation. A Morse oscillator having three sets of vibronic states is used, as an example, to evaluate the selectivity and efficiency of two-photon excitation. We show that a vibrational mode can be selectively excited with high efficiency by the correlated photons, without phase manipulation or pulse-shaping techniques. This can be achieved by controlling the quantum correlation so that the photon pair concurrently has two pulse widths, namely, a temporally narrow width and a spectrally narrow width. Though this concurrence is seemingly contradictory, we can create such a photon pair by tailoring the quantum correlation between two photons.

  18. Near infrared two-photon excitation cross-sections of voltage-sensitive dyes.

    Science.gov (United States)

    Fisher, Jonathan A N; Salzberg, Brian M; Yodh, Arjun G

    2005-10-15

    Microscopy based on voltage-sensitive dyes has proven effective for revealing spatio-temporal patterns of neuronal activity in vivo and in vitro. Two-photon microscopy using voltage-sensitive dyes offers the possibility of wide-field visualization of membrane potential on sub-cellular length scales, hundreds of microns below the tissue surface. Very little information is available, however, about the utility of voltage-sensitive dyes for two-photon imaging purposes. Here we report on measurements of two-photon fluorescence excitation cross-sections for nine voltage-sensitive dyes in a solvent, octanol, intended to simulate the membrane environment. Ultrashort light pulses from a Ti:sapphire laser were used for excitation from 790 to 960 nm, and fluorescein dye was used as a calibration standard. Overall, dyes RH795, RH421, RH414, di-8-ANEPPS, and di-8-ANEPPDHQ had the largest two-photon excitation cross-sections ( approximately 15 x 10(-50)cm4 s photon(-1)) in this wavelength region and are therefore potentially useful for two-photon microscopy. Interestingly, di-8-ANEPPDHQ, a chimera constructed from the potentiometric dyes RH795 and di-8-ANEPPS, exhibited larger cross-sections than either of its constituents.

  19. Highly selective population of two excited states in nonresonant two-photon absorption

    Institute of Scientific and Technical Information of China (English)

    Zhang Hui; Zhang Shi-An; Sun Zhen-Rong

    2011-01-01

    A nonresonant two-photon absorption process can be manipulated by tailoring the ultra-short laser pulse.In this paper,we theoretically demonstrate a highly selective population of two excited states in the nonresonant two-photon absorption process by rationally designing a spectral phase distribution.Our results show that one excited state is maximally populated while the other state population is widely tunable from zero to the maximum value.We believe that the theoretical results may play an important role in the selective population of a more complex nonlinear process comprising nonresonant two-photon absorption,such as resonance-mediated(2+1)-three-photon absorption and (2+1)-resonant multiphoton ionization.

  20. Sub-diffraction positioning of a two-photon excited and optically trapped quantum dot

    DEFF Research Database (Denmark)

    Jauffred, L.; Kyrsting, A.; Christensen, Eva Arnspang;

    2014-01-01

    Colloidal quantum dots are luminescent long-lived probes that can be two-photon excited and manipulated by a single laser beam. Therefore, quantum dots can be used for simultaneous single molecule visualization and force manipulation using an infra-red laser. Here, we show that even a single opti...

  1. Two-photon excitation spectra of Cr3 :K2NaScF6

    Science.gov (United States)

    Bartram, R. H.; Wein, G. R.; Hamilton, D. S.; Sliwczuk, U.; Rinzler, A. G.

    Two-photon excitation (TPE) spectra of Cr3+:K2NaScF6 exhibit unexpected features including a forbidden transition, extended progressions, a split zero-phonon line and anomalous polarization anisotropy. These features are explained by departures from standard approximations.

  2. Diagnostics of MCF plasmas using Lyman-{alpha} fluorescence excited by one or two photons

    Energy Technology Data Exchange (ETDEWEB)

    Voslamber, D

    1998-11-01

    Laser-induced Lyman-{alpha} fluorescence of the hydrogen isotopes is investigated with regard to diagnostic applications in magnetically confined fusion plasmas. A formal analysis is presented for two excitation schemes: one-photon and Doppler-free two-photon excitation. The analysis includes estimates of the expected experimental errors arising from the photon noise and from the sensitivity of the observed fluorescence signals to variations of the plasma and laser parameters. Both excitation schemes are suitable primarily for application in the plasma edge, but even in the plasma bulk of large machines they can still be applied in combination with a diagnostic neutral beam. The two-photon excitation scheme is particularly attractive because it involves absorption spectra that are resolved within the Doppler width. This implies a large diagnostic potential and in particular offers a way to measure the deuterium-tritium fuel mix in fusion reactors. (author) 37 refs.

  3. Direct two-photon excitation of isomeric transition in thorium-229 nucleus

    CERN Document Server

    Romanenko, V I; Yatsenko, L P; Romanenko, A V; Litvinov, A N; Kazakov, G A

    2012-01-01

    A possibility of the two-photon excitation of an isomeric state in a nucleus of thorium-229 has been discussed. The fluorescence intensity of the excitation is demonstrated to be identical for the irradiation of nuclei with either monochromatic light or polychromatic radiation consisting of a sequence of short light pulses of the same intensity. The two-photon excitation of Th^{3+} ion in an electromagnetic trap with a focused laser beam with a wavelength of about 320 nm and power of 100 mW can lead to the absorption saturation, at which the fluorescence emission with the frequency of the transition in a nucleus is maximal. In crystals doped with Th^{4+} to a concentration of about 10^{18} cm^{-3} and irradiated with a laser radiation 10 W in power, the emission of several photons per second with a wavelength of about 160 nm becomes possible.

  4. Nonlinear processes upon two-photon interband picosecond excitation of PbWO4 crystal

    Science.gov (United States)

    Lukanin, V. I.; Karasik, A. Ya

    2016-09-01

    A new experimental method is proposed to study the dynamics of nonlinear processes occurring upon two-photon interband picosecond excitation of a lead tungstate crystal and upon its excitation by cw probe radiation in a temporal range from several nanoseconds to several seconds. The method is applied to the case of crystal excitation by a sequence of 25 high-power picosecond pulses with a wavelength of 523.5 nm and 633-nm cw probe radiation. Measuring the probe beam transmittance during crystal excitation, one can investigate the influence of two-photon interband absorption and the thermal nonlinearity of the refractive index on the dynamics of nonlinear processes in a wide range of times (from several nanoseconds to several seconds). The time resolution of the measuring system makes it possible to distinguish fast and slow nonlinear processes of electronic or thermal nature, including the generation of a thermal lens and thermal diffusion. An alternative method is proposed to study the dynamics of induced absorption transformation and, therefore, the dynamics of the development of nonlinear rocesses upon degenerate two-photon excitation of the crystal in the absence of external probe radiation.

  5. Stepwise Two-Photon-Induced Fast Photoswitching via Electron Transfer in Higher Excited States of Photochromic Imidazole Dimer.

    Science.gov (United States)

    Kobayashi, Yoichi; Katayama, Tetsuro; Yamane, Takuya; Setoura, Kenji; Ito, Syoji; Miyasaka, Hiroshi; Abe, Jiro

    2016-05-11

    Stepwise two-photon excitations have been attracting much interest because of their much lower power thresholds compared with simultaneous two-photon processes and because some stepwise two-photon processes can be initiated by a weak incoherent excitation light source. Here we apply stepwise two-photon optical processes to the photochromic bridged imidazole dimer, whose solution instantly changes color upon UV irradiation and quickly reverts to the initial color thermally at room temperature. We synthesized a zinc tetraphenylporphyrin (ZnTPP)-substituted bridged imidazole dimer, and wide ranges of time-resolved spectroscopic studies revealed that a ZnTPP-linked bridged imidazole dimer shows efficient visible stepwise two-photon-induced photochromic reactions upon excitation at the porphyrin moiety. The fast photoswitching property combined with stepwise two-photon processes is important not only for the potential for novel photochromic materials that are sensitive to the incident light intensity but also for fundamental photochemistry using higher excited states.

  6. Electric field allowed molecular transitions for one and two photon excitation microscopy.

    Science.gov (United States)

    Mondal, Partha Pratim; Diaspro, Alberto

    2008-07-01

    We propose an excitation technique for observing single and two photon excitation in those molecules for which such transitions are forbidden by the selection rules. This is possible by the application of an external electric field that perturbs the molecular orbitals, thereby resulting in a significant shift of energy levels. Such a shift of energy levels may bring those levels in resonance with the radiation field which is normally forbidden by selection rules. Further, parity of the these states may significantly improve the emission process. The external electric field results in the mixing of excited (short lifetime) and metastable states (long lifetime), thus reducing the lifetime of metastable (or near metastable) states. This may provide an effective channel for allowing transition from the metastable states. An application of electric field may result in the excitation of poorly excitable biomolecules. This excitation technique may find applications in single- and multi-photon fluorescence microscopy, bioimaging and optical devices.

  7. Two-photon excited fluorescence microendoscopic imaging using a GRIN lens

    Science.gov (United States)

    Yan, Wei; Peng, Xiao; Lin, Danying; Wang, Qi; Gao, Jian; Zhou, Jie; Ye, Tong; Qu, Junle; Niu, Hanben

    2015-03-01

    With the rapid development of life sciences, there is an increasing demand for intravital fluorescence imaging of small animals. However, large dimensions and limited working distances of objective lenses in traditional fluorescence microscopes have limited the imaging applications mostly to superficial tissues. To overcome this disadvantage, researchers have developed the graded-index (GRIN) probes with small diameters for imaging internal organs of small animals in a minimally invasive fashion. Here, we present the development of a fluorescence endoscopic imaging system based on a GRIN lens using two-photon excitation. Experimental results showed that this system could perform dynamic fluorescence microendoscopic imaging and monitor the blood flow in anesthetized living mice using two-photon excitation.

  8. In vitro imaging of thyroid tissues using two-photon excited fluorescence and second harmonic generation.

    Science.gov (United States)

    Huang, Zufang; Li, Zuanfang; Chen, Rong; Lin, Juqiang; Li, Yongzeng; Li, Chao

    2010-08-01

    To evaluate the feasibility of two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) imaging to discriminate the normal, nodular goiter and papillary cancerous thyroid tissue. In total, 45 fresh thyroid specimens (normal, 15; nodular goiter, 12; and papillary cancerous, 18) from 31 subjects were directly imaged by the TPEF and SHG combination method. The microstructure of follicle and collagen structure in thyroid tissue were clearly identified, morphologic changes between normal, nodular goiter, and papillary cancerous thyroid tissue were well characterized by using two-photon excitation fluorescence. SHG imaging of the collagen matrix also revealed the differences between normal and abnormal. Our preliminary study suggests that the TPEF and SHG combination method might be a useful tool in revealing pathologic changes in thyroid tissue.

  9. Adiabatic rapid passage two-photon excitation of a Rydberg atom

    CERN Document Server

    Kuznetsova, Elena; Malinovskaya, Svetlana A

    2015-01-01

    We considered the two-photon adiabatic rapid passage excitation of a single atom from the ground to a Rydberg state. Three schemes were analyzed: both pump and Stokes fields chirped and pulsed, only the pump field is chirped, and only the pump field is pulsed and chirped while the Stokes field is continuous wave (CW). In all three cases high transfer efficiencies $>99\\%$ were achieved for the experimentally realizable Rabi frequencies and the pulse durations of the fields.

  10. [Intensity loss of two-photon excitation fluorescence microscopy images of mouse oocyte chromosomes].

    Science.gov (United States)

    Zhao, Feng-Ying; Wu, Hong-Xin; Chen, Die-Yan; Ma, Wan-Yun

    2014-07-01

    As an optical microscope with high resolution, two-photon excitation (TPE) fluorescence microscope is widely used in noninvasive 3D optical imaging of biological samples. Compared with confocal laser scanning microscope, TPE fluorescence microscope provides a deeper detecting depth. In spite of that, the image quality of sample always declines as the detecting depth increases when a noninvasive 3D optical imaging of thicker samples is performed. Mouse oocytes with a large diameter, which play an important role in clinical and biological fields, have obvious absorption and scattering effects. In the present paper, we performed compensation for two-photon fluorescence images of mouse oocyte chromosomes. Using volume as a parameter, the attenuation degree of these chromosomes was also studied. The result of our data suggested that there exists a severe axial intensity loss in two-photon microscopic images of mouse oocytes due to the absorption and scattering effects. It is necessary to make compensation for these images of mouse oocyte chromosomes obtained from two-photon microscopic system. It will be specially needed in studying the quantitative three-dimensional information of mouse oocytes.

  11. Polarised two-photon excitation of quantum well excitons for manipulation of optically pumped terahertz lasers

    Energy Technology Data Exchange (ETDEWEB)

    Slavcheva, G., E-mail: gsk23@bath.ac.uk [Blackett Laboratory, Imperial College London, Prince Consort Road, London SW7 2AZ (United Kingdom); Kavokin, A.V., E-mail: A.Kavokin@soton.ac.uk [School of Physics and Astronomy, University of Southampton, Highfield, Southampton SO17 1BJ (United Kingdom); Spin Optics Laboratory, St. Petersburg State University, 1, Ulyanovskaya 198504 (Russian Federation)

    2014-11-15

    Optical pumping of excited exciton states in a semiconductor quantum well embedded in a microcavity is a tool for realisation of ultra-compact terahertz (THz) lasers based on stimulated optical transition between excited (2p) and ground (1s) exciton state. We show that the probability of two-photon absorption by a 2p-exciton is strongly dependent on the polarisation of both pumping photons. Five-fold variation of the threshold power for terahertz lasing by switching from circular to co-linear pumping is predicted. We identify photon polarisation configurations for achieving maximum THz photon generation quantum efficiency.

  12. Single & Two-photon Excited Fluorescence of Two New Compounds with 2-Benzothiazolyl as Electron Acceptor

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    Two new D-π-A type compounds, where electron-donor D is tertiary amino group, electron-acceptor A is 2-benzothiazolyl and π is two conjugated styryl units, have been synthesized.They are named as trans, trans-2-{4-[4-(N, N-diethylamino)styryl]styryl}-1, 3-benzothiazole and trans, trans-2-{4-[4-(N, N-diphenylamino)styryl]styryl}-1, 3-benzothiazole.Both compounds show strong two-photon excited fluorescence in yellow-orange region when excited by a femtosecond laser at 800 nm.

  13. [Two-photon excitation fluorescence of 5-ALA induced PpIX in DHL cells].

    Science.gov (United States)

    Huang, Zu-Fang; Chen, Rong; Li, Yong-Zeng; Chen, Guan-Nan; Chen, Xian-Ling; Feng, Shang-Yuan; Jia, Pei-Min

    2008-11-01

    Two-photon fluorescence microscopy is a novel imaging technique, which is primarily sensitive to a specimen's response coming from an in-focus plane, thus has low photo-bleaching and photo-damage to biological samples. 5-ALA induced production of PpIX in DHL cells was excited by 820 nm femtosecond laser; two-photon excitation fluorescence of single cell was obtained in Lambda mode of laser scanning confocal microscope. The specific fluorescence intensity of PpIX which accumulated in DHL cells was measured at 2, 4 and 10 mmol x L(-1) concentration of 5-ALA with different incubation time, which reflected the kinetics of 5-ALA accumulated in DHL cells. Accumulation of PpIX in DHL cells was a dynamic change process. Biphasic alterations of PpIX accumulation were noted: PpIX content enhanced with the increasing time and reached the maximal value around 3 h, however PpIX content decreased in the subsequent incubation time. Results indicate that two-photon fluorescence based on laser scanning microscope can be a useful technology for studying the kinetics of 5-ALA induced PpIX production in DHL cells and other leukemia cells.

  14. Enhanced two-photon excited fluorescence from imaging agents using true thermal light

    Science.gov (United States)

    Jechow, Andreas; Seefeldt, Michael; Kurzke, Henning; Heuer, Axel; Menzel, Ralf

    2013-12-01

    Two-photon excited fluorescence (TPEF) is a standard technique in modern microscopy, but is still affected by photodamage to the probe. It has been proposed that TPEF can be enhanced using entangled photons, but this has proven challenging. Recently, it was shown that some features of entangled photons can be mimicked with thermal light, which finds application in ghost imaging, subwavelength lithography and metrology. Here, we use true thermal light from a superluminescent diode to demonstrate TPEF that is enhanced compared to coherent light, using two common fluorophores and luminescent quantum dots, which suit applications in imaging and microscopy. We find that the TPEF rate is directly proportional to the measured degree of second-order coherence, as predicted by theory. Our results show that photon bunching in thermal light can be exploited in two-photon microscopy, with the photon statistic providing a new degree of freedom.

  15. Polarization and spectral characteristics of the two-photon luminescence from colloidal gold nanoparticles excited by tunable laser radiation

    Energy Technology Data Exchange (ETDEWEB)

    Yashunin, D. A., E-mail: yashuninda@yandex.ru; Korytin, A. I.; Stepanov, A. N. [Russian Academy of Sciences, Institute of Applied Physics (Russian Federation)

    2015-12-15

    We have experimentally studied two-photon luminescence from a colloidal solution of spherical gold nanoparticles by tuning the wavelength of the exciting radiation. The measured polarization and spectral characteristics of the two-photon luminescence signal show that the observed nonlinear optical response is determined by the dimers present in the solution with a concentration of a few percent of total nanoparticle number.

  16. Coherent blue emission generated by Rb two-photon excitation using diode and femtosecond lasers

    Science.gov (United States)

    Lopez, Jesus P.; Moreno, Marco P.; de Miranda, Marcio H. G.; Vianna, Sandra S.

    2017-04-01

    The coherent blue light generated in rubidium vapor due to the combined action of an ultrashort pulse train and a continuous wave diode laser is investigated. Each step of the two-photon transition 5S-5P{}3/2-5D is excited by one of the lasers, and the induced coherence between the 5S and 6P{}3/2 states is responsible for generating the blue beam. Measurements of the excitation spectrum reveal the frequency comb structure and allow us to identify the resonant modes responsible for inducing the nonlinear process. Further, each resonant mode excites a different group of atoms, making the process selective in atomic velocity. The signal dependency on the atomic density is characterized by a sharp growth and a rapid saturation. We also show that for high intensity of the diode laser, the Stark shift at resonance causes the signal suppression observed at low atomic density.

  17. Nanoshells for in vivo imaging using two-photon excitation microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Gao Liang; Nammalvar, Vengadesan [Department of Bioengineering, Rice University, Houston, TX 77005 (United States); Vadakkan, Tegy J, E-mail: lg3@rice.edu, E-mail: venkyn@rice.edu [Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, TX 77030 (United States)

    2011-09-07

    Gold nanoshells have been intensively investigated and applied to various biomedical fields because of their flexible optical tunability and biological compatibility. They hold great potential to serve as luminescent contrast agents excitable with near-infrared (NIR) lasers. In this paper, we describe the development of nanoshells with a peak of plasmon resonance at 800 nm and their subsequent use for in vivo blood vessel imaging using two-photon excitation microscopy at an excitation wavelength of 750 nm. We were able to image single nanoshell particles in blood vessels and generate optical contrast for blood vessel structure using luminescent signals. These results confirm the feasibility of engineering nanoshells with controlled optical properties for single-particle-based in vivo imaging.

  18. Nanoshells for in vivo imaging using two-photon excitation microscopy.

    Science.gov (United States)

    Gao, Liang; Vadakkan, Tegy J; Nammalvar, Vengadesan

    2011-09-07

    Gold nanoshells have been intensively investigated and applied to various biomedical fields because of their flexible optical tunability and biological compatibility. They hold great potential to serve as luminescent contrast agents excitable with near-infrared (NIR) lasers. In this paper, we describe the development of nanoshells with a peak of plasmon resonance at 800 nm and their subsequent use for in vivo blood vessel imaging using two-photon excitation microscopy at an excitation wavelength of 750 nm. We were able to image single nanoshell particles in blood vessels and generate optical contrast for blood vessel structure using luminescent signals. These results confirm the feasibility of engineering nanoshells with controlled optical properties for single-particle-based in vivo imaging.

  19. Imaging zebrafish embryos by two-photon excitation time-lapse microscopy.

    Science.gov (United States)

    Carvalho, Lara; Heisenberg, Carl-Philipp

    2009-01-01

    The zebrafish is a favorite model organism to study tissue morphogenesis during development at a subcellular level. This largely results from the fact that zebrafish embryos are transparent and thus accessible to various imaging techniques, such as confocal and two-photon excitation (2PE) microscopy. In particular, 2PE microscopy has been shown to be useful for imaging deep cell layers within the embryo and following tissue morphogenesis over long periods. This chapter describes how to use 2PE microscopy to study morphogenetic movements during early zebrafish embryonic development, providing a general blueprint for its use in zebrafish.

  20. One- and two-photon excited fluorescence lifetimes and anisotropy decays of green fluorescent proteins.

    OpenAIRE

    2000-01-01

    We have used one- (OPE) and two-photon (TPE) excitation with time-correlated single-photon counting techniques to determine time-resolved fluorescence intensity and anisotropy decays of the wild-type Green Fluorescent Protein (GFP) and two red-shifted mutants, S65T-GFP and RSGFP. WT-GFP and S65T-GFP exhibited a predominant approximately 3 ns monoexponential fluorescence decay, whereas for RSGFP the main lifetimes were approximately 1.1 ns (main component) and approximately 3.3 ns. The anisotr...

  1. Resonant transfer of one- and two-photon excitations in quantum dot-bacteriorhodopsin complexes

    Science.gov (United States)

    Krivenkov, V. A.; Samokhvalov, P. S.; Bilan, R. S.; Chistyakov, A. A.; Nabiev, I. R.

    2017-01-01

    Light-sensitive protein bacteriorhodopsin (BR), which is capable of electrical response upon exposure to light, is a promising material for photovoltaics and optoelectronics. However, the rather narrow absorption spectrum of BR does not allow achieving efficient conversion of the light energy in the blue and infrared spectral regions. This paper summarizes the results of studies showing the possibility of extending the spectral region of the BR function by means of the Förster resonance energy transfer (FRET) from CdSe/ZnS quantum dots (QDs), which have a broad spectrum of one-photon absorption and a large twophoton absorption cross section (TPACS), to BR upon one- and two-photon excitation. In particular, it is shown that, on the basis of QDs and BR-containing purple membranes, it is possible to create electrostatically associated bio-nano hybrid systems in which FRET is implemented. In addition, the large TPACS of QDs, which is two orders of magnitude larger than those of BR and organic dyes, opens up a means for selective two-photon excitation of synthesized bio-nano hybrid complexes. On the basis of the results of this work, the spectral region in which BR converts the light energy into electrical energy can be extended from the UV to near-IR region, creating new opportunities for the use of this material in photovoltaics and optoelectronics.

  2. Increasing efficiency of two-photon excited fluorescence and second harmonic generation using ultrashort pulses

    Science.gov (United States)

    Tang, Shuo; Krasieva, Tatiana B.; Chen, Zhongping; Tempea, Gabriel; Tromberg, Bruce J.

    2006-02-01

    Multiphoton microscopy (MPM) has become an important tool for high-resolution and non-invasive imaging in biological tissues. However, the efficiencies of two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) are relatively low because of their nonlinear nature. Therefore, it is critical to optimize laser parameters for most efficient excitation of MPM. Reducing the pulse duration can increase the peak intensity of excitation and thus potentially increase the excitation efficiency. In this paper, a multiphoton microscopy system using a 12 fs Ti:Sapphire laser is reported. With adjustable dispersion pre-compensation, the pulse duration at the sample location can be varied from 400 fs to sub-20 fs. The efficiencies of TPEF and SHG are studied for the various pulse durations, respectively. Both TPEF and SHG are found to increase proportionally to the inverse of the pulse duration for the entire tested range. To transmit most of the SHG and TPEF signals, the spectral transmission widow of the detection optics needs to be carefully considered. Limitation from phase-matching in SHG generation is not significant because the effective interaction length for SHG is less than 10 μm at the focal depth of the objectives. These results are important in improving MPM excitation efficiency using ultrashort pulses. MPM images from human artery wall are also demonstrated.

  3. Identification of calcifications in intracranial neoplasms using two photon excitation fluorescence microscopy

    Science.gov (United States)

    Lin, Peihua; Wang, Xingfu; Wu, Zanyi; Fang, Na; Li, Lianhuang; Kang, Dezhi; Chen, Jianxin

    2016-10-01

    Calcifications within brain tumors may be an indicator of a relatively long survival because a long time is required for the formation of calcium deposits, and may present a novel biomarker associated with response and improved outcome of therapy. In this paper, we describe the use of two-photon excitation fluorescent (TPEF) microscopy combined second harmonic generation (SHG) microscopy for high-resolution imaging that can be applied in identification of intratumoral calcifications. Our results demonstrate that the calcification has stronger TPEF signal than the area around it and the emission spectra shows the difference between the two areas clearly. The TPEF image of calcified region corresponds well with the corresponding H&E stained image. In this work, we present that the label-free imaging technique is able to distinguish the calcified mass lesions in intracranial neoplasms reliably.

  4. GPC light shaper for speckle-free one- and two-photon contiguous pattern excitation

    DEFF Research Database (Denmark)

    Bañas, Andrew Rafael; Palima, Darwin; Villangca, Mark Jayson;

    2014-01-01

    Generalized Phase Contrast (GPC) is an efficient method for generating speckle-free contiguous optical distributions useful in diverse applications such as static beam shaping, optical manipulation and recently, for excitation in two-photon optogenetics. To fully utilize typical Gaussian lasers......, such as a circle and different rectangles commonly used in industrial or commercial applications. We also show simple and efficient beam shaping of arbitrary shapes geared towards biophotonics research and other contemporary applications. Optimized GPC configurations consistently give ~84% efficiency and ~3x...... in such applications, we analytically derive conditions for photon efficient light shaping with GPC. When combined with the conditions for optimal contrast developed in previous works, our analysis further simplifies GPCx2019;s implementation. The results of our analysis are applied to practical illumination shapes...

  5. Quantitative optical biomarkers of lung cancer based intrinsic two-photon excited fluorescence signal

    Science.gov (United States)

    Li, Jingwen; Zhan, Zhenlin; Lin, Hongxin; Zuo, Ning; Zhu, Xiaoqin; Xie, Shusen; Chen, Jianxin; Zhuo, Shuangmu

    2016-10-01

    Alterations in the elastic fibers have been implicated in lung cancer. However, the label-free, microscopic imaging of elastic fibers in situ remains a major challenge. Here, we present the use of intrinsic two-photon excited fluorescence (TPEF) signal as a novel means for quantification of the elastic fibers in intact fresh human lung tissues. We obtained the TPEF images of elastic fibers from ex vivo the human lung tissues. We found that three features, including the elastic fibers area, the elastic fibers orientation, the elastic fibers structure, provide the quantitative identification of lung cancer and the direct visual cues for cancer versus non-cancer areas. These results suggest that the TPEF signal can be used as the label-free optical biomarkers for rapid clinical lung diagnosis and instant image-guided surgery.

  6. One- and Two-photon Excited Fluorescence of Zinc(Ⅱ), Cadmium(Ⅱ) Complexes Containing Phenothiazine Ligand

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    A new ligand, 10-ethylphenothiazinyl - 3 - yl - methylene thiosemicarbazon (HL) and its complexes ML2 (M=Zn2+, Cd2+), which exhibit intensive two-photon excited (TPE) fluorescence at 800 nm laser pulses in femtosecond regime, were synthesized and characterized.The measured power dependence of the fluorescence signals provided direct evidence for TPE.All of them exhibited a large two-photon absorptive cross section and, more importantly from the application point of view, high photochemical/photothermal stability.

  7. Imaging-guided two-photon excitation-emission-matrix measurements of human skin tissues

    Science.gov (United States)

    Yu, Yingqiu; Lee, Anthony M. D.; Wang, Hequn; Tang, Shuo; Zhao, Jianhua; Lui, Harvey; Zeng, Haishan

    2012-07-01

    There are increased interests on using multiphoton imaging and spectroscopy for skin tissue characterization and diagnosis. However, most studies have been done with just a few excitation wavelengths. Our objective is to perform a systematic study of the two-photon fluorescence (TPF) properties of skin fluorophores, normal skin, and diseased skin tissues. A nonlinear excitation-emission-matrix (EEM) spectroscopy system with multiphoton imaging guidance was constructed. A tunable femtosecond laser was used to vary excitation wavelengths from 730 to 920 nm for EEM data acquisition. EEM measurements were performed on excised fresh normal skin tissues, seborrheic keratosis tissue samples, and skin fluorophores including: NADH, FAD, keratin, melanin, collagen, and elastin. We found that in the stratum corneum and upper epidermis of normal skin, the cells have large sizes and the TPF originates from keratin. In the lower epidermis, cells are smaller and TPF is dominated by NADH contributions. In the dermis, TPF is dominated by elastin components. The depth resolved EEM measurements also demonstrated that keratin structure has intruded into the middle sublayers of the epidermal part of the seborrheic keratosis lesion. These results suggest that the imaging guided TPF EEM spectroscopy provides useful information for the development of multiphoton clinical devices for skin disease diagnosis.

  8. Highly Efficient and Excitation Tunable Two-Photon Luminescence Platform For Targeted Multi-Color MDRB Imaging Using Graphene Oxide

    Science.gov (United States)

    Pramanik, Avijit; Fan, Zhen; Chavva, Suhash Reddy; Sinha, Sudarson Sekhar; Ray, Paresh Chandra

    2014-08-01

    Multiple drug-resistance bacteria (MDRB) infection is one of the top three threats to human health according to the World Health Organization (WHO). Due to the large penetration depth and reduced photodamage, two-photon imaging is an highly promising technique for clinical MDRB diagnostics. Since most commercially available water-soluble organic dyes have low two-photon absorption cross-section and rapid photobleaching tendency, their applications in two-photon imaging is highly limited. Driven by the need, in this article we report extremely high two-photon absorption from aptamer conjugated graphene oxide (σ2PA = 50800 GM) which can be used for highly efficient two-photon fluorescent probe for MDRB imaging. Reported experimental data show that two-photon photoluminescence imaging color, as well as luminescence peak position can be tuned from deep blue to red, just by varying the excitation wavelength without changing its chemical composition and size. We have demonstrated that graphene oxide (GO) based two-photon fluorescence probe is capable of imaging of multiple antibiotics resistance MRSA in the first and second biological transparency windows using 760-1120 nm wavelength range.

  9. Optical control of cardiac cell excitability based on two-photon infrared absorption of AzoTAB

    CERN Document Server

    Shcherbakov, D; Erofeev, I; Astafiev, A

    2014-01-01

    Recent studies of AzoTAB activity in excitable cell cultures have shown that this substance is able to control excitability depending on isomer, cis or trans, predominating in the cellular membrane. Control of isomerization can be performed noninvasively by UV-visual radiation. At the same time it is well-known that azobenezenes can be effectively transformed from one isomer into another by two-photon absorption. Current work is devoted to the study of trans-AzoTAB two-photon transformation in aqueous solution and inside primal neonatal contractive rat cardiomyocytes. In accordance with results obtained Azo-TAB can be used as a probe for two-photon optical control of cardiac excitability.

  10. Multicolor excitation two-photon microscopy: in vivo imaging of cells and tissues

    Science.gov (United States)

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

    2010-02-01

    Two-photon microscopy based on endogenous fluorescence provides non-invasive imaging of living biological system. Reduced nicotinamide adenine dinucleotide (NADH), flavin adenine dinucleotide (FAD), keratin, collagen and elastin are the endogenous fluorophores widely used as the contrast agents for imaging metabolism and morphology of living cells and tissue. The fluorescence of tryptophan, a kind of essential amino acid, conveys the information on cellular protein content, structure and microenvironment. However, it can't be effectively excited by the commonly used Ti:sapphire femtosecond laser. Because each endogenous fluorophore provides limited information, it is desirable to simultaneously excite fluorescence from as many fluorophores as possible to obtain accurate biochemical and morphological information on biomedical samples. In this study, we demonstrate that the supercontinuum generation from a photonic crystal fiber (PCF) excited by an ultrafast source can be used to excite multiple endogenous nonlinear optical signals simultaneously. By employing the spectral lifetime detection capability, this technology provides a unique approach to sense the fine structure, protein distribution and cellular metabolism of cells and tissues in vivo. In particular, with application of acetic acid, a safe contrast agent used for detection cervical cancer for many years, the tryptophan signals reveal cellular morphology and even cell-cell junctions clearly. Moreover, it was found that the pH value dependent lifetime of tryptophan fluorescence could provide the qualitative information on the gradient of pH value in epithelial tissue. Finally, we will demonstrate the potential of our multi-color TPEF microscopy to investigate the early development of cancer in epithelial tissue.

  11. Induced structural defects in Ti-doped ZnO and its two-photon-excitation

    Science.gov (United States)

    Martínez Julca, Milton A.; Rivera, Ivonnemary; Santillan Mercado, Jaime; Sierra, Heidy; Perales-Pérez, Oscar

    2016-03-01

    ZnO is a well-known luminescent material that reacts with light to generate free radicals enabling its use in cancer treatment by Photodynamic Therapy (PDT). Unfortunately, up to know, the photo-excitation of ZnO-based materials' requires excitation with ultraviolet light, which limits their biomedical applications. In this regard, this work investigates the effect of Ti species incorporation into the lattice of ZnO nanoparticles (NPs) with the aim of improving the corresponding optical properties and enabling the two-photoexcitation with 690nm-light (near infrared light). A modified polyol-based route was used to synthesize pure and Ti-doped (9% at.) ZnO NPs. X-ray diffraction confirmed the formation of ZnO-wurtzite whereas Scanning Electron Microscopy confirmed the formation of monodispersed 100-nm NPs. Raman Spectroscopy measurements evidenced the presence of zinc interstitials (Zni) and oxygen vacancies (VO) in the host oxide strcuture. Asynthesized NPs were excited using the technique of two-photon fluorescence microscopy (TPFM). The photoluminescence (PL) spectra generated from the analysis of TPFM images revealed a high emission peak presence in the green region (555 nm) that was assigned to VO. Also, a weak but noticeable band at 420 nm was detected, which is attributed to electron transition from the shallow donor level of Zni to the valence band. These PL transitions will favor triplet states formation necessary to yield cytotoxic reactive oxygen species. Furthermore, the presence of the PL peaks confirmed the Ti-ZnO NPs capacity to be excited by 690-nm light, thus, opening new possibilities for this NPs to be used in lightinduced bio-medical applications.

  12. Spectral and lifetime endomicroscopic measurements using one and two-photon excitation

    Science.gov (United States)

    Ibrahim, A.; Poulon, F.; Zanello, M.; Habert, R.; Varlet, P.; Devaux, B.; Kudlinski, A.; Abi Haidar, D.

    2017-02-01

    Current surgical biopsy needs several days for the analysis process to be finished. Anatomopathologists provide analysis reports to the surgeon a few days after the surgical intervention, which makes it a lengthy decision making practice. In addition, the lack of precise guidance often leads to inaccuracies in the selection of tissue regions for biopsy and so necessitates repeating the operation sometimes. Our project aims at reducing this time as well as patient discomfort. In this context, we propose to develop a multimodal nonlinear endomicroscope providing several means of contrast. Among these contrast that are useful in the detection of tumor regions, we note imaging by linear and non-linear fluorescence, by second and third harmonic generation and by reflectance. In addition, this technique allows fluorescence lifetime and spectral measurements. Our endomicroscopic system is based on a new homemade customized double-clad photonic crystal fiber (DC-PCF). Finally, this double-clad micro structured optical fiber insures visible and near infrared excitation. This system was tested by measuring fluorescence lifetime and the spectral shape of a fixed tumoral brain sample in one and two photon excitations.

  13. Stimulated emission of free excitons in Cd{sub 1-x}Mn{sub x}Te under nonresonant two-photon excitation

    Energy Technology Data Exchange (ETDEWEB)

    Jang, J.I. [Department of Physics and Astronomy, Northwestern University, 2145 Sheridan Road, Evanston, IL 60208 (United States)], E-mail: joon-jang@northwestern.edu; Mani, S.; Ketterson, J.B. [Department of Physics and Astronomy, Northwestern University, 2145 Sheridan Road, Evanston, IL 60208 (United States); Park, H.Y. [Department of Semiconductor Applications, Ulsan College, San 29 Mugeo Dong, Ulsan 680-749 (Korea, Republic of)], E-mail: hypark@mail.uc.ac.kr

    2008-08-25

    We report on free excitons coexisting with exciton magnetic polarons (EMPs) in bulk semimagnetic semiconductors of Cd{sub 1-x}Mn{sub x}Te for 0.04{<=}x{<=}0.36 at 2 K under nonresonant two-photon excitation. This two-photon excitation not only generates free excitons but also more efficiently creates EMPs compared with ordinary one-photon excitation. Stimulated emission from free excitons is demonstrated under strong two-photon excitation.

  14. Two-color two-photon excited fluorescence of indole: Determination of wavelength-dependent molecular parameters

    Energy Technology Data Exchange (ETDEWEB)

    Herbrich, Sebastian; Al-Hadhuri, Tawfik; Gericke, Karl-Heinz, E-mail: k.Gericke@tu-bs.de [Institut für Physikalische und Theoretische Chemie, TU Braunschweig, Hans-Sommer-Straße 10, 38106 Braunschweig (Germany); Shternin, Peter S., E-mail: pshternin@gmail.com; Vasyutinskii, Oleg S., E-mail: osv@pms.ioffe.ru [Ioffe Institute, Politekhnicheskaya 26, 194021 St. Petersburg (Russian Federation); St. Petersburg Polytechnic University, Politekhnicheskaya 29, St. Petersburg 195251 (Russian Federation); Smolin, Andrey G. [Ioffe Institute, Politekhnicheskaya 26, 194021 St. Petersburg (Russian Federation)

    2015-01-14

    We present a detailed study of two-color two-photon excited fluorescence in indole dissolved in propylene glycol. Femtosecond excitation pulses at effective wavelengths from 268 to 293.33 nm were used to populate the two lowest indole excited states {sup 1}L{sub a} and {sup 1}L{sub b} and polarized fluorescence was then detected. All seven molecular parameters and the two-photon polarization ratio Ω containing information on two-photon absorption dynamics, molecular lifetime τ{sub f}, and rotation correlation time τ{sub rot} have been determined from experiment and analyzed as a function of the excitation wavelength. The analysis of the experimental data has shown that {sup 1}L{sub b}–{sup 1}L{sub a} inversion occurred under the conditions of our experiment. The two-photon absorption predominantly populated the {sup 1}L{sub a} state at all excitation wavelengths but in the 287–289 nm area which contained an absorption hump of the {sup 1}L{sub b} state 0-0 origin. The components of the two-photon excitation tensor S were analyzed giving important information on the principal tensor axes and absorption symmetry. The results obtained are in a good agreement with the results reported by other groups. The lifetime τ{sub f} and the rotation correlation time τ{sub rot} showed no explicit dependence on the effective excitation wavelength. Their calculated weighted average values were found to be τ{sub f} = 3.83 ± 0.14 ns and τ{sub rot} = 0.74 ± 0.06 ns.

  15. Collimated Blue and Infrared Beams Generated by Two-Photon Excitation in Rb Vapor

    CERN Document Server

    Sell, J F; DePaola, B D; Knize, R J

    2013-01-01

    Utilizing two-photon excitation in hot Rb vapor we demonstrate the generation of collimated optical fields at 420 nm and 1324 nm. Input laser beams at 780 nm and 776 nm enter a heated Rb vapor cell collinear and circularly polarized, driving Rb atoms to the $5D_{5/2}$ state. Under phase-matching conditions coherence among the $5S_{1/2}\\rightarrow 5P_{3/2}\\rightarrow 5D_{5/2} \\rightarrow 6P_{3/2}$ transitions produces a blue (420 nm) beam by four-wave mixing. We also observe a forward and backward propagating IR (1324 nm) beam, due to cascading decays through the $6S_{1/2}\\rightarrow 5P_{1/2}$ states. Power saturation of the generated beams is investigated by scaling the input powers to greater than 200 mW, resulting in a coherent blue beam of 9.1 mW power, almost an order of magnitude larger than previously achieved. We measure the dependences of both beams in relation to the Rb density, the frequency detuning between Rb ground state hyperfine levels, and the input laser intensities.

  16. Two-photon excited fluorescence lifetime imaging and spectroscopy of melanins in vitro and in vivo

    Science.gov (United States)

    Krasieva, Tatiana B.; Stringari, Chiara; Liu, Feng; Sun, Chung-Ho; Kong, Yu; Balu, Mihaela; Meyskens, Frank L.; Gratton, Enrico; Tromberg, Bruce J.

    2013-03-01

    Changes in the amounts of cellular eumelanin and pheomelanin have been associated with carcinogenesis. The goal of this work is to develop methods based on two-photon-excited-fluorescence (TPEF) for measuring relative concentrations of these compounds. We acquire TPEF emission spectra (λex=1000 nm) of melanin in vitro from melanoma cells, hair specimens, and in vivo from healthy volunteers. We find that the pheomelanin emission peaks at approximately 615 to 625 nm and eumelanin exhibits a broad maximum at 640 to 680 nm. Based on these data we define an optical melanin index (OMI) as the ratio of fluorescence intensities at 645 and 615 nm. The measured OMI for the MNT-1 melanoma cell line is 1.6±0.22 while the Mc1R gene knockdown lines MNT-46 and MNT-62 show substantially greater pheomelanin production (OMI=0.5±0.05 and 0.17±0.03, respectively). The measured values are in good agreement with chemistry-based melanin extraction methods. In order to better separate melanin fluorescence from other intrinsic fluorophores, we perform fluorescence lifetime imaging microscopy of in vitro specimens. The relative concentrations of keratin, eumelanin, and pheomelanin components are resolved using a phasor approach for analyzing lifetime data. Our results suggest that a noninvasive TPEF index based on spectra and lifetime could potentially be used for rapid melanin ratio characterization both in vitro and in vivo.

  17. Two-Photon Excitation STED Microscopy with Time-Gated Detection.

    Science.gov (United States)

    Coto Hernández, Iván; Castello, Marco; Lanzanò, Luca; d'Amora, Marta; Bianchini, Paolo; Diaspro, Alberto; Vicidomini, Giuseppe

    2016-01-13

    We report on a novel two-photon excitation stimulated emission depletion (2PE-STED) microscope based on time-gated detection. The time-gated detection allows for the effective silencing of the fluorophores using moderate stimulated emission beam intensity. This opens the possibility of implementing an efficient 2PE-STED microscope with a stimulated emission beam running in a continuous-wave. The continuous-wave stimulated emission beam tempers the laser architecture's complexity and cost, but the time-gated detection degrades the signal-to-noise ratio (SNR) and signal-to-background ratio (SBR) of the image. We recover the SNR and the SBR through a multi-image deconvolution algorithm. Indeed, the algorithm simultaneously reassigns early-photons (normally discarded by the time-gated detection) to their original positions and removes the background induced by the stimulated emission beam. We exemplify the benefits of this implementation by imaging sub-cellular structures. Finally, we discuss of the extension of this algorithm to future all-pulsed 2PE-STED implementationd based on time-gated detection and a nanosecond laser source.

  18. Tuning Ag29 nanocluster light emission from red to blue with one and two-photon excitation

    Science.gov (United States)

    Russier-Antoine, Isabelle; Bertorelle, Franck; Hamouda, Ramzi; Rayane, Driss; Dugourd, Philippe; Sanader, Željka; Bonačić-Koutecký, Vlasta; Brevet, Pierre-François; Antoine, Rodolphe

    2016-01-01

    We demonstrate that the tuning of the light emission from red to blue in dihydrolipoic acid (DHLA) capped Ag29 nanoclusters can be trigged with one and two photon excitations. The cluster stoichiometry was determined with mass spectrometry and found to be Ag29(DHLA)12. In a detailed optical investigation, we show that these silver nanoclusters exhibit a strong red photoluminescence visible to the naked eye and characterized by a quantum yield of nearly ~2% upon one-photon excitation. In the nonlinear optical (NLO) study of the properties of the clusters, the two-photon excited fluorescence spectra were recorded and their first hyperpolarizability obtained. The two-photon absorption cross-section at ~800 nm for Ag29(DHLA)12 is higher than 104 GM and the hyperpolarizability is 106 × 10-30 esu at the same excitation wavelength. The two-photon excited fluorescence spectrum appears strongly blue-shifted as compared to the one-photon excited spectrum, displaying a broad band between 400 and 700 nm. The density functional theory (DFT) provides insight into the structural and electronic properties of Ag29(DHLA)12 as well as into interplay between metallic subunit or core and ligands which is responsible for unique optical properties.We demonstrate that the tuning of the light emission from red to blue in dihydrolipoic acid (DHLA) capped Ag29 nanoclusters can be trigged with one and two photon excitations. The cluster stoichiometry was determined with mass spectrometry and found to be Ag29(DHLA)12. In a detailed optical investigation, we show that these silver nanoclusters exhibit a strong red photoluminescence visible to the naked eye and characterized by a quantum yield of nearly ~2% upon one-photon excitation. In the nonlinear optical (NLO) study of the properties of the clusters, the two-photon excited fluorescence spectra were recorded and their first hyperpolarizability obtained. The two-photon absorption cross-section at ~800 nm for Ag29(DHLA)12 is higher than 104

  19. ``Entangled'' free-induction decay in CdS crystal under two-photon excitation by two crossed laser beams

    Science.gov (United States)

    Leontiev, A. V.; Lobkov, V. S.; Mitrofanova, T. G.; Shmelyov, A. G.; Samartsev, V. V.

    2012-09-01

    A new method of two-photon excitation of femtosecond signals of ``entangled'' free induction decay (EFID) by two crossed 790-nm laser beams in a CdS crystal at room temperature has been realized for the first time. This ``entangled'' (through the wave vectors) coherent response appears only in the case when the photons involved to the process of two-photon excitation of the sample belong to the different laser beams. This technique allows one to separate the EFID signal from the exciting femtosecond pulses and to vary the response wavelength by varying the angle between their wave vectors. The most optimal case occurs when the angle between the wave vectors of exciting pulses as well as the angle between each of these wave vectors and that of the response is equal to 60°.

  20. Magneto-Photoluminescence Based on Two-Photon Excitation in Lanthanide-Doped Up-Conversion Crystal Particles.

    Science.gov (United States)

    Xu, Hengxing; Qin, Wei; Li, Mingxing; Wu, Ting; Hu, Bin

    2017-04-01

    Experimental studies on magneto-photoluminescence based on two-photon excitation in up-conversion Y2 O2 S:Er, Yb crystal particles are reported. It is found that the up-conversion photoluminescence generated by two-photon excitation exhibits magnetic field effects at room temperature, leading to a two-photon excitation-induced magneto-photoluminescence, when the two-photon excitation exceeds the critical intensity. By considering the spin selection rule in electronic transitions, it is proposed that spin-antiparallel and spin-parallel transition dipoles with spin mixing are accountable for the observed magneto-photoluminescence. Specifically, the two-photon excitation generates spin-antiparallel electric dipoles between (4) S3/2 -(4) I15/2 in Er(3+) ions. The antiparallel spins are conserved by exchange interaction within dipoles. When the photoexcitation exceeds the critical intensity, the Coulomb screening can decrease the exchange interaction. Consequently, the spin-orbital coupling can partially convert the antiparallel dipoles into parallel dipoles, generating a spin mixing. Eventually, the populations between antiparallel and parallel dipoles reach an equilibrium established by the competition between exchange interaction and spin-orbital coupling. Applying a magnetic field can break the equilibrium by disturbing spin mixing through introducing spin precessions, changing the spin populations on antiparallel and parallel dipoles and leading to the magneto-photoluminescence. Therefore, spin-dependent transition dipoles present a convenient mechanism to realize magneto-photoluminescence in multiphoton up-conversion crystal particles. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Peptide backbone orientation and dynamics in spider dragline silk and two-photon excitation in nuclear magnetic and quadrupole resonance

    Energy Technology Data Exchange (ETDEWEB)

    Eles, P.T

    2005-07-01

    In the first part of the dissertation, spider dragline silk is studied by solid state NMR techniques. The dependence of NMR frequency on molecular orientation is exploited using the DECODER experiment to determine the orientation of the protein backbone within the silk fibre. Practical experimental considerations require that the silk fibres be wound about a cylindrical axis perpendicular to the external magnetic field, complicating the reconstruction of the underlying orientation distribution and necessitating the development of numerical techniques for this purpose. A two-component model of silk incorporating static b-sheets and polyglycine II helices adequately fits the NMR data and suggests that the b-sheets are well aligned along the silk axis (20 FWHM) while the helices are poorly aligned (68 FWHM). The effects of fibre strain, draw rate and hydration on orientation are measured. Measurements of the time-scale for peptide backbone motion indicate that when wet, a strain-dependent fraction of the poorly aligned component becomes mobile. This suggests a mechanism for the supercontraction of silk involving latent entropic springs that undergo a local strain-dependent phase transition, driving supercontraction. In the second part of this dissertation a novel method is developed for exciting NMR and nuclear quadrupole resonance (NQR) by rf irradiation at multiple frequencies that sum to (or differ by) the resonance frequency. This is fundamentally different than traditional NMR experiments where irradiation is applied on-resonance. With excitation outside the detection bandwidth, two-photon excitation allows for detection of free induction signals during excitation, completely eliminating receiver dead-time. A theoretical approach to describing two-photon excitation is developed based on average Hamiltonian theory. An intuition for two-photon excitation is gained by analogy to the coherent absorption of multiple photons requiring conservation of total energy and

  2. Two-photon excited fluorescence spectroscopy and imaging of melanin in vitro and in vivo

    Science.gov (United States)

    Krasieva, Tatiana B.; Liu, Feng; Sun, Chung-Ho; Kong, Yu; Balu, Mihaela; Meyskens, Frank L.; Tromberg, Bruce J.

    2012-03-01

    The ability to detect early melanoma non-invasively would improve clinical outcome and reduce mortality. Recent advances in two-photon excited fluorescence (TPEF) in vivo microscopy offer a powerful tool in early malignant melanoma diagnostics. The goal of this work was to develop a TPEF optical index for measuring relative concentrations of eumelanin and pheomelanin since ex vivo studies show that changes in this ratio have been associated with malignant transformation. We acquired TPEF emission spectra (λex=1000 nm) of melanin from several specimens, including human hair, malignant melanoma cell lines, and normal melanocytes and keratinocytes in different skin layers (epidermis, papillary dermis) in five healthy volunteers in vivo. We found that the pheomelanin emission peaks at around 620 nm and is blue-shifted from the eumelanin with broad maximum at 640-680nm. We defined "optical melanin index" (OMI) as a ratio of fluorescence signal intensities measured at 645 nm and 615nm. The measured OMI for a melanoma cell line MNT-1 was 1.6+/-0.2. The MNT-46 and MNT-62 lines (Mc1R gene knockdown) showed an anticipated change in melanins production ratio and had OMI of 0.55+/-0.05 and 0.17+/-0.02, respectively, which strongly correlated with HPLC data obtained for these lines. Average OMI measured for basal cells layers (melanocytes and keratinocytes) in normal human skin type I, II-III (not tanned and tanned) in vivo was 0.5, 1.05 and 1.16 respectively. We could not dependably detect the presence of pheomelanin in highly pigmented skin type V-VI. These data suggest that a non-invasive TPEF index could potentially be used for rapid melanin ratio characterization both in vitro and in vivo, including pigmented lesions.

  3. Photosensitizer-doped conjugated polymer nanoparticles with high cross-sections for one- and two-photon excitation.

    Science.gov (United States)

    Grimland, Jennifer L; Wu, Changfeng; Ramoutar, Ria R; Brumaghim, Julia L; McNeill, Jason

    2011-04-01

    We report a novel nanoparticle that is promising for photodynamic therapy applications, which consists of a π-conjugated polymer doped with a singlet oxygen photosensitizer. The nanoparticles exhibit highly efficient collection of excitation light due to the large excitation cross-section of the polymer. A quantum efficiency of singlet oxygen production of 0.5 was determined. Extraordinarily large two-photon excitation cross-sections were determined, indicating promise for near infrared multiphoton photodynamic therapy. Gel electrophoresis of DNA after near-UV irradiation in the presence of nanoparticles indicated both purine base and backbone DNA damage.

  4. Two-photon excitation with pico-second fluorescence lifetime imaging to detect nuclear association of flavanols.

    Science.gov (United States)

    Mueller-Harvey, Irene; Feucht, Walter; Polster, Juergen; Trnková, Lucie; Burgos, Pierre; Parker, Anthony W; Botchway, Stanley W

    2012-03-16

    Two-photon excitation enabled for the first time the observation and measurement of excited state fluorescence lifetimes from three flavanols in solution, which were ~1.0 ns for catechin and epicatechin, but <45 ps for epigallocatechin gallate (EGCG). The shorter lifetime for EGCG is in line with a lower fluorescence quantum yield of 0.003 compared to catechin (0.015) and epicatechin (0.018). In vivo experiments with onion cells demonstrated that tryptophan and quercetin, which tend to be major contributors of background fluorescence in plant cells, have sufficiently low cross sections for two-photon excitation at 630 nm and therefore do not interfere with detection of externally added or endogenous flavanols in Allium cepa or Taxus baccata cells. Applying two-photon excitation to flavanols enabled 3-D fluorescence lifetime imaging microscopy and showed that added EGCG penetrated the whole nucleus of onion cells. Interestingly, EGCG and catechin showed different lifetime behaviour when bound to the nucleus: EGCG lifetime increased from <45 to 200 ps, whilst catechin lifetime decreased from 1.0 ns to 500 ps. Semi-quantitative measurements revealed that the relative ratios of EGCG concentrations in nucleoli associated vesicles: nucleus: cytoplasm were ca. 100:10:1. Solution experiments with catechin, epicatechin and histone proteins provided preliminary evidence, via the appearance of a second lifetime (τ(2)=1.9-3.1 ns), that both flavanols may be interacting with histone proteins. We conclude that there is significant nuclear absorption of flavanols. This advanced imaging using two-photon excitation and biophysical techniques described here will prove valuable for probing the intracellular trafficking and functions of flavanols, such as EGCG, which is the major flavanol of green tea.

  5. Two-Photon Study on the Electronic Interactions between the First Excited Singlet States in Carotenoid-Tetrapyrrole Dyads

    Energy Technology Data Exchange (ETDEWEB)

    Liao, Pen-Nan [Technische Universitat Braunschweig (Germany); Pillai, Smitha [Arizona State Univ., Tempe, AZ (United States); Gust, Devens [Arizona State Univ., Tempe, AZ (United States); Moore, Thomas A. [Arizona State Univ., Tempe, AZ (United States); Moore, Ana L. [Arizona State Univ., Tempe, AZ (United States); Walla, Peter J. [Technische Universitat Braunschweig (Germany)

    2011-03-22

    Electronic interactions between the first excited states (S1) of carotenoids (Car) of different conjugation lengths (8-11 double bonds) and phthalocyanines (Pc) in different Car-Pc dyad molecules were investigated by two-photon spectroscopy and compared with Car S1-chlorophyll (Chl) interactions in photosynthetic light harvesting complexes (LHCs). The observation of Chl/Pc fluorescence after selective two-photon excitation of the Car S1 state allowed sensitive monitoring of the flow of energy between Car S1 and Pc or Chl. It is found that two-photon excitation excites to about 80% to 100% exclusively the carotenoid state Car S1 and that only a small fraction of direct tetrapyrrole two-photon excitation occurs. Amide-linked Car-Pc dyads in tetrahydrofuran demonstrate a molecular gear shift mechanism in that effective Car S1 → Pc energy transfer is observed in a dyad with 9 double bonds in the carotenoid, whereas in similar dyads with 11 double bonds in the carotenoid, the Pc fluorescence is strongly quenched by Pc → Car S1 energy transfer. In phenylamino-linked Car-Pc dyads in toluene extremely large electronic interactions between the Car S1 state and Pc were observed, particularly in the case of a dyad in which the carotenoid contained 10 double bonds. This observation together with previous findings in the same system provides strong evidence for excitonic Car S1-Pc Qy interactions. Very similar results were observed with photosynthetic LHC II complexes in the past, supporting an important role of such interactions in photosynthetic down-regulation.

  6. The controlled excitation of forbidden transitions in the two-photon spectrum of strontium by using collisions and electric fields

    Science.gov (United States)

    Philip, G.; Connerade, J.-P.

    2007-11-01

    We present experimental results involving controlled configuration mixing in two-photon spectroscopy of highly-excited states by exploiting a weak external electric field and collisions. The method has allowed new extensions to high members of the two-photon forbidden J = 3 odd-parity 5snf 1F 3 and the J = 0, even-parity 5sns 1S 0 Rydberg series of neutral strontium to be observed. We achieve resonant two-photon transverse excitation of a high density atomic jet by using a narrow bandwidth tunable dye laser in a heat pipe setup with sensitive ionization detection. Experimental term values are extended for the 5sns 1S 0 series up to n = 46. By suitable exploitation of the composition and pressure of the buffer gases in conjunction with the electric field strength in the excitation region and the exciting laser beam intensity we have also extended observations up to n = 44 for the 5snf 1F 3 series and up to n = 46 for the 5snp 1P 1 series. Our results demonstrate a novel and remarkably simple experimental method to access high Rydberg states to which transitions are forbidden from the ground state by parity and other selection rules.

  7. Fluorenyl porphyrins for combined two-photon excited fluorescence and photosensitization

    Science.gov (United States)

    Mongin, Olivier; Hugues, Vincent; Blanchard-Desce, Mireille; Merhi, Areej; Drouet, Samuel; Yao, Dandan; Paul-Roth, Christine

    2015-04-01

    The two-photon absorption (2PA), the luminescence and the photosensitization properties of porphyrin-cored fluorenyl dendrimers and meso-substituted fluorenylporphyrin monomer, dimer and trimer are described. In comparison with model tetraphenylporphyrin, these compounds combine enhanced (non-resonant) 2PA cross-sections in the near infrared and enhanced fluorescence quantum yields, together with maintained singlet oxygen generation quantum yields. 'Semi-disconnection' between fluorenyl groups and porphyrins (i.e. direct meso substitution) proved to be more efficient than non-conjugated systems (based on efficient FRET between fluorenyl antennae and porphyrins). These results are of interest for combined two-photon imaging and photodynamic therapy.

  8. Photo-redox activated drug delivery systems operating under two photon excitation in the near-IR.

    Science.gov (United States)

    Guardado-Alvarez, Tania M; Devi, Lekshmi Sudha; Vabre, Jean-Marie; Pecorelli, Travis A; Schwartz, Benjamin J; Durand, Jean-Olivier; Mongin, Olivier; Blanchard-Desce, Mireille; Zink, Jeffrey I

    2014-05-07

    We report the design and synthesis of a nano-container consisting of mesoporous silica nanoparticles with the pore openings covered by "snap-top" caps that are opened by near-IR light. A photo transducer molecule that is a reducing agent in an excited electronic state is covalently attached to the system. Near IR two-photon excitation causes inter-molecular electron transfer that reduces a disulfide bond holding the cap in place, thus allowing the cargo molecules to escape. We describe the operation of the "snap-top" release mechanism by both one- and two-photon activation. This system presents a proof of concept of a near-IR photoredox-induced nanoparticle delivery system that may lead to a new type of photodynamic drug release therapy.

  9. Two-photon excited highly polarized and directional upconversion emission from slab organic crystals

    NARCIS (Netherlands)

    Fang, Hong-Hua; Chen, Qi-Dai; Yang, Jie; Xia, Hong; Ma, Yu-Guang; Wang, Hai-Yu; Sun, Hong-Bo; Fang, Honghua

    2010-01-01

    Effective upconversion emission from an organic crystal of cyano-substituted oligo (p-phenylenevinylene) (CNDPASDB) based on two-photon absorption is presented. Frequency upconverted cavityless lasing, or amplified spontaneous emission, from the crystal pumped by a femtosecond laser of 800 nm was ob

  10. Widefield Two-Photon Excitation without Scanning: Live Cell Microscopy with High Time Resolution and Low Photo-Bleaching.

    Directory of Open Access Journals (Sweden)

    Rumelo Amor

    Full Text Available We demonstrate fluorescence imaging by two-photon excitation without scanning in biological specimens as previously described by Hwang and co-workers, but with an increased field size and with framing rates of up to 100 Hz. During recordings of synaptically-driven Ca(2+ events in primary rat hippocampal neurone cultures loaded with the fluorescent Ca(2+ indicator Fluo-4 AM, we have observed greatly reduced photo-bleaching in comparison with single-photon excitation. This method, which requires no costly additions to the microscope, promises to be useful for work where high time-resolution is required.

  11. Widefield two-photon excitation without scanning: live cell microscopy with high time resolution and low photo-bleaching

    CERN Document Server

    Amor, Rumelo; Robb, Gillian; Wilson, Louise; Rahman, Nor Zaihana Abdul; Dempster, John; Amos, William Bradshaw; Bushell, Trevor J; McConnell, Gail

    2015-01-01

    We demonstrate fluorescence imaging by two-photon excitation without scanning in biological specimens as previously described by Hwang and co-workers, but with an increased field size and with framing rates of up to 100 Hz. During recordings of synaptically-driven Ca$^{2+}$ events in primary rat neurone cultures loaded with the fluorescent Ca$^{2+}$ indicator Fluo-4 AM, we have observed greatly reduced photo-bleaching in comparison with single-photon excitation. This method, which requires no costly additions to the microscope, promises to be useful for work where high time-resolution is required.

  12. Energy transfer in aminonaphthalimide-boron-dipyrromethene (BODIPY) dyads upon one- and two-photon excitation: applications for cellular imaging.

    Science.gov (United States)

    Collado, Daniel; Remón, Patricia; Vida, Yolanda; Najera, Francisco; Sen, Pratik; Pischel, Uwe; Perez-Inestrosa, Ezequiel

    2014-03-01

    Aminonaphthalimide-BODIPY energy transfer cassettes were found to show very fast (kEET ≈ 10(10)-10(11) s(-1) and efficient BODIPY fluorescence sensitization. This was observed upon one- and two-photon excitation, which extends the application range of the investigated bichromophoric dyads in terms of accessible excitation wavelengths. In comparison with the direct excitation of the BODIPY chromophore, the two-photon absorption cross-section δ of the dyads is significantly incremented by the presence of the aminonaphthalimide donor [δ ≈ 10 GM for the BODIPY versus 19-26 GM in the dyad at λ(exc)=840 nm; 1 GM (Goeppert-Mayer unit)=10(-50) cm(4) smolecule(-1) photon-(1)]. The electronic decoupling of the donor and acceptor, which is a precondition for the energy transfercassette concept, was demonstrated by time-dependent density functional theory calculations. The applicability of the new probes in the one- and twophoton excitation mode was demonstrated in a proof-of-principle approach in the fluorescence imaging of HeLa cells. To the best of our knowledge, this is the first demonstration of the merging of multiphoton excitation with the energy transfer cassette concept for a BODIPY-containing dyad.

  13. Temperature-dependent excitonic photoluminescence Excited by Two-Photon Absorption in Perovskite CsPbBr3 Quantum Dots

    CERN Document Server

    Wei, Ke; Xu, Zhongjie; Shen, Chao; Cheng, Xiangai; Jiang, Tian

    2016-01-01

    Recently lead halide nanocrystals (quantum dots) have been reported with potential for photovoltaic and optoelectronic applications due to their excellent luminescent properties. Herein excitonic photoluminescence (PL) excited by two-photon absorption in perovskite CsPbBr3 quantum dots (QDs) have been studied across a broad temperature range from 80K to 380K. Two-photon absorption has been investigated with absorption coefficient up to 0.085 cm/GW at room temperature. Moreover, the photoluminescence excited by two-photon absorption shows a linear blue-shift (0.25meV/K) below temperature of ~220K and turned steady with fluctuation below 1nm (4.4meV) for higher temperature up to 380K. These phenomena are distinctly different from general red-shift of semiconductor and can be explained by the competition between lattice expansion and electron-phonon couplling.Our results reveal the strong nonlinear absorption and temperature-independent chromaticity in a large temperature range from 220K to 380K in the CsPbX3 QD...

  14. Two-photon excited fluorescence of intrinsic fluorophores enables label-free assessment of adipose tissue function

    Science.gov (United States)

    Alonzo, Carlo Amadeo; Karaliota, Sevasti; Pouli, Dimitra; Liu, Zhiyi; Karalis, Katia P.; Georgakoudi, Irene

    2016-08-01

    Current methods for evaluating adipose tissue function are destructive or have low spatial resolution. These limit our ability to assess dynamic changes and heterogeneous responses that occur in healthy or diseased subjects, or during treatment. Here, we demonstrate that intrinsic two-photon excited fluorescence enables functional imaging of adipocyte metabolism with subcellular resolution. Steady-state and time-resolved fluorescence from intracellular metabolic co-factors and lipid droplets can distinguish the functional states of excised white, brown, and cold-induced beige fat. Similar optical changes are identified when white and brown fat are assessed in vivo. Therefore, these studies establish the potential of non-invasive, high resolution, endogenous contrast, two-photon imaging to identify distinct adipose tissue types, monitor their functional state, and characterize heterogeneity of induced responses.

  15. Selective Two-Photon Absorptive Resonance Femtosecond-Laser Electronic-Excitation Tagging (STARFLEET) Velocimetry in Flow and Combustion Diagnostics

    Science.gov (United States)

    Jiang, Naibo; Halls, Benjamin R.; Stauffer, Hans U.; Roy, Sukesh; Danehy, Paul M.; Gord, James R.

    2016-01-01

    Selective Two-Photon Absorptive Resonance Femtosecond-Laser Electronic-Excitation Tagging (STARFLEET), a non-seeded ultrafast-laser-based velocimetry technique, is demonstrated in reactive and non-reactive flows. STARFLEET is pumped via a two-photon resonance in N2 using 202.25-nm 100-fs light. STARFLEET greatly reduces the per-pulse energy required (30 µJ/pulse) to generate the signature FLEET emission compared to the conventional FLEET technique (1.1 mJ/pulse). This reduction in laser energy results in less energy deposited in the flow, which allows for reduced flow perturbations (reactive and non-reactive), increased thermometric accuracy, and less severe damage to materials. Velocity measurements conducted in a free jet of N2 and in a premixed flame show good agreement with theoretical velocities and further demonstrate the significantly less-intrusive nature of STARFLEET.

  16. One- and two-photon spectroscopy of highly excited states of alkali-metal atoms on helium nanodroplets.

    Science.gov (United States)

    Pifrader, Alexandra; Allard, Olivier; Auböck, Gerald; Callegari, Carlo; Ernst, Wolfgang E; Huber, Robert; Ancilotto, Francesco

    2010-10-28

    Alkali-metal atoms captured on the surface of superfluid helium droplets are excited to high energies (≈3 eV) by means of pulsed lasers, and their laser-induced-fluorescence spectra are recorded. We report on the one-photon excitation of the (n+1)p←ns transition of K, Rb, and Cs (n=4, 5, and 6, respectively) and on the two-photon one-color excitation of the 5d←5s transition of Rb. Gated-photon-counting measurements are consistent with the relaxation rates of the bare atoms, hence consistent with the reasonable expectation that atoms quickly desorb from the droplet and droplet-induced relaxation need not be invoked.

  17. Assessing topographic cutaneous autofluorescence variation using fluorescence UV and visible excitation emission matrix (EEM) spectroscopy

    Science.gov (United States)

    Zhao, Jianhua; Zandi, Soodabeh; Feng, Florina; Zeng, Haishan; McLean, David I.; Lui, Harvey

    2011-03-01

    Cutaneous autofluorescence properties were systematically studied using fluorescence excitation emission matrix spectroscopy. Twenty-six healthy subjects with a mean age of 34 (range 21-74) participated in this study. The fluorescence of major skin fluorophores such as tryptophan, collagen, elastin and NADH could be readily identified. On average, facial skin shows strong tryptophan and measurable porphyrin fluorescence; the palm and nail show strong tryptophan and keratin fluorescence. These results demonstrate that regional topographic variations exist not only in the amount of fluorescence but also in the relative distribution of fluorophores in normal skin. Moreover this provides a basis for future interpretation of autofluorescence in diseased skin.

  18. Optically Pumped Atomic Rubidium Lasers: Two-Photon and Exciplex Excitation Mechanisms

    Science.gov (United States)

    Gallagher, Jeffrey E.

    The Doppler-broadened two-photon absorption (TPA) cross-section for the 52S1/2 → 52 D5/2 transition in Rb is measured using direct absorption methods. The selection rule |DeltaF| ≤ 2 applied to both isotopes yields 17 transitions in 3 Doppler limited lines. A detailed model of the intensity profile was also developed to account for a focused Gaussian beam (with an M2 value of 1.09) propagating through a two-photon absorption medium. A peak absorbance of 24% was observed for an intensity of 6.28 kWcm2 at the focus, a Rb density of 4.6x1015 cm-3 , and a path length of 15 cm. Alkali concentrations from 1.61 - 8.52x1015 cm -3 were monitored in the far wing of the D 2 line. Extracting the hyperfine-broadened TPA cross-section from 87 test configurations, while varying the pump power, alkali concentration and focal length, yielded an error-weighted average of 6.75x10^-21 cm4W with a standard deviation of 3.61x10-21 cm4W. This cross-section is sufficient for a pulsed dye laser to bleach the pump transition in the Two-Photon Pumped Alkali Laser (TPAL) that lases at 420 nm and 5.2 microm. Optically pumped atomic rubidium lasers pumped in the blue satellite of the D2 line from the ground Rb-Ar or Rb-Kr collision pair to the dissociative B2S+1/2 state produce laser emission at 780.2 nm. Lasing is achieved for pump wavelengths of 752.3 to greater than 760 nm for the Rb-Ar system and 757.1 -- 760.4 nm for the Rb-Kr system. Slope efficiencies increase with both Rb and Ar concentrations and exceed 0.25% using a heat pipe configuration. The gain is very high with photon build-up times of 1--3.7 ns. Laser induced heating and subsequent condensation of alkali vapor in the heat pipe configuration currently limits operation to less than 2500 Torr.

  19. Phasor analysis of multiphoton spectral images distinguishes autofluorescence components of in vivo human skin

    NARCIS (Netherlands)

    Fereidouni, F.; Bader, A.N.; Colonna, A.; Gerritsen, H.C.

    2014-01-01

    Skin contains many autofluorescent components that can be studied using spectral imaging. We employed a spectral phasor method to analyse two photon excited auto-fluorescence and second harmonic generation images of in vivo human skin. This method allows segmentation of images based on spectral feat

  20. Femtosecond correlated photon echo in CdS crystal under two-photon excitation by two pairs of crossed laser beams

    Science.gov (United States)

    Samartsev, V. V.; Leontiev, A. V.; Mitrofanova, T. G.

    2015-07-01

    We consider the possibility of observing a femtosecond correlated photon echo (FCPE) under two-photon excitation of CdS crystal by two pairs of crossed laser beams. The peculiarities of FCPE signals and their possible applications are discussed.

  1. Comparing temporally-focused GPC and CGH for two-photon excitation and optogenetics in turbid media

    DEFF Research Database (Denmark)

    Villangca, Mark Jayson; Bañas, Andrew Rafael; Aabo, Thomas

    2013-01-01

    Inherent inhomogeneity in turbid media not only hinders imaging but also projection of arbitrary light patterns for excitation or optical manipulation. In this work we compare two of the most popular phase modulation-based techniques in beam shaping. The Generalized Phase Contrast (GPC) method uses...... a 4f setup that directly converts phase information to intensity. The GPC method has been used with temporal focusing for excitation in two-photon optogenetics [1-3]. The computer generated hologram (CGH) is also used to generate arbitrary light patterns and has been used for optical manipulation...... and fabrication because of its high diffraction efficiency and axial confinement. We model the effect of the turbid media as a phase randomization process. We compare the quality and asses the degradation of the projected light pattern for both techniques as it propagates in the turbid media....

  2. Relaxation Process of Excitonic Molecules in CuCl under the Two-Photon Resonant Excitation. II. Transverse Relaxation

    Science.gov (United States)

    Itoh, Tadashi; Katohno, Takashi; Kirihara, Toshio; Ueta, Masayasu

    1984-02-01

    Under the off-resonant excitation at the slightly higher energy side of the giant two-photon absorption band (GTA) for the direct generation of excitonic molecules (EM) in CuCl, new narrow emission bands designated as XT and XL have been found in the energy region of the MT and ML broad bands, respectively. Their photon energies increase with the decrease of the pump photon energy from the higher-energy side of the resonance and finally, at the on-resonant excitation, their bands merge into MT0 and ML0 bands, respectively, previously reported by Mita et al. Based on the detailed studies on these bands, it is found that there exists a certain kind of transverse relaxation process which acts on the EM just after their generation by the GTA and brings about the X emission as a hot luminescence.

  3. Multiphoton microscopic imaging of adipose tissue based on second-harmonic generation and two-photon excited fluorescence.

    Science.gov (United States)

    Huang, Zufang; Zhuo, Shuangmu; Chen, Jianxin; Chen, Rong; Jiang, Xingshan

    2008-01-01

    The fresh adipose tissue was investigated by the use of multiphoton microscopy (MPM) based on two-photon excited fluorescence and second-harmonic generation (SHG). Microstructure of collagen and adipose cells in the adipose tissue is clearly imaged at a subcellular level with the excitation light wavelengths of 850 and 730 nm, respectively. The emission spectrum of collagen SHG signal and NADH and FAD fluorescence signal can also be obtained, which can be used to quantify the content of collagen and adipose cells and reflect the degree of pathological changes when comparing normal tissue with abnormal adipose tissue in the same condition. The results indicate that MPM has the potential to be applied to investigate the adipose tissue and can be used in the research field of lipid and connective tissues.

  4. Characterization of scintillating CaWO{sub 4} crystals for the CRESST experiment using two-photon excitation

    Energy Technology Data Exchange (ETDEWEB)

    Hampf, Raphael; Dandl, Thomas; Muenster, Andrea; Oberauer, Lothar; Roth, Sabine; Schoenert, Stefan; Ulrich, Andreas [Physik-Department and Excellence Cluster Universe, Technische Universitaet Muenchen, D-85747 Garching (Germany)

    2016-07-01

    In the CRESST experiment for direct dark matter search, phonon and photon signals from cryogenic CaWO{sub 4} crystals are used to search for WIMP-induced nuclear recoil events. We present a novel table-top setup in which the scintillation of CaWO{sub 4} is induced by 0.7 ns laser pulses of 355 nm wavelength. The excitation occurs via two-photon absorption in the bulk material. The scintillation light is observed by time resolved optical spectroscopy. By varying the focusing of the laser-beam the excitation density can be made high enough to study quenching effects due to exciton-exciton annihilation. This allows to perform experiments to test models for the quenching factors of different ionizing projectiles in CaWO{sub 4} which are used to identify these projectiles on an event by event basis.

  5. Two-photon excitation spectroscopy of Cr3+:K2NaScF6 elpasolite: II. Theoretical models

    Science.gov (United States)

    Bartram, R. H.; Wein, G. R.; Hamilton, D. S.

    2001-03-01

    Two-photon excitation (TPE) spectra of Cr3+:K2NaScF6, excited by a Raman-shifted, Nd:YAG-pumped tunable dye laser, exhibit several unexpected features. A weak TPE spectrum of the symmetry-forbidden 4A2g→ 4T2g transition is observed without a zero-phonon line. The symmetry-allowed TPE spectrum of the 4A2g→ 4T1ag transition has a multi-phonon side band with anomalously extended vibrational progressions, and an anomalously weak, split zero-phonon line with anomalous polarization anisotropy. These observations are explained, respectively, in terms of theoretical models involving phonon assistance, departures from the closure approximation that permit electron-lattice coupling in intermediate states and a low-temperature phase transition involving librational instability. Hypothetical line-shape simulations are compared with observed TPE spectra.

  6. Single particle tracking through highly scattering media with multiplexed two-photon excitation

    Science.gov (United States)

    Perillo, Evan; Liu, Yen-Liang; Liu, Cong; Yeh, Hsin-Chih; Dunn, Andrew K.

    2015-03-01

    3D single-particle tracking (SPT) has been a pivotal tool to furthering our understanding of dynamic cellular processes in complex biological systems, with a molecular localization accuracy (10-100 nm) often better than the diffraction limit of light. However, current SPT techniques utilize either CCDs or a confocal detection scheme which not only suffer from poor temporal resolution but also limit tracking to a depth less than one scattering mean free path in the sample (typically validated our microscope by tracking (1) fluorescent nanoparticles in a prescribed motion inside gelatin gel (with 1% intralipid) and (2) labeled single EGFR complexes inside skin cancer spheroids (at least 8 layers of cells thick) for ~10 minutes. Furthermore we discuss future capabilities of our multiplexed two-photon microscope design, specifically to the extension of (1) simultaneous multicolor tracking (i.e. spatiotemporal co-localization analysis) and (2) FRET studies (i.e. lifetime analysis). The high resolution, high depth penetration, and multicolor features of this microscope make it well poised to study a variety of molecular scale dynamics in the cell, especially related to cellular trafficking studies with in vitro tumor models and in vivo.

  7. Highly efficient and two-photon excited stimulated Rayleigh-Bragg scattering in organic solutions

    Energy Technology Data Exchange (ETDEWEB)

    He, Guang S., E-mail: gshe@buffalo.edu; Prasad, Paras N. [The Institute for Lasers, Photonics and Biophotonics, State University of New York at Buffalo, Buffalo, New York 14260-3000 (United States); Kannan, Ramamurthi; Tan, Loon-Seng [Air Force Research Laboratory, Materials and Manufacturing Directorate, AFRL/RX, Wright-Patterson AFB, Ohio 45433-7750 (United States)

    2015-07-21

    The properties of backward stimulated Rayleigh-Bragg scattering (SRBS) in three highly two-photon active AF-chromophores solutions in tetrahydrofuran (THF) have been investigated using 816-nm and 8-ns pump laser beam. The nonlinear reflectivity R, spectral structure, temporal behavior, and phase-conjugation capability of the backward SRBS output have been measured, respectively. Under the same experimental condition, the pump threshold for SRBS in three solution samples can be significantly (∼one order of magnitude) lower than that for stimulated Brillouin scattering (SBS) in the pure solvent (THF). With the optimized concentration value and at a moderate pump energy (∼1.5 mJ) level, the measured nonlinear reflectivity was R ≥ 35% for the 2 cm-long solution sample, while for the SBS from a pure solvent sample of the same length was R ≈ 4.7%. The peculiar features of very low pump threshold, no spectral shift, tolerant pump spectral linewidth requirement (≤1 cm{sup −1}), and phase-conjugation capability are favorable for those nonlinear photonics applications, such as highly efficiency phase-conjugation reflectors for high-brightness laser oscillator/amplifier systems, special imaging through turbid medium, self-adaptive remote optical sensing, as well as for optical rangefinder and lidar systems.

  8. Two-photon- photoluminescence excitation spectroscopy of single quantum-dots

    CERN Document Server

    Benny, Y; Poem, E; Khatsevitch, S; Gershoni, D; Petroff, P M

    2011-01-01

    We present experimental and theoretical study of single semiconductor quantum dots excited by two non-degenerate, resonantly tuned variably polarized lasers. The first laser is tuned to excitonic resonances. Depending on its polarization it photogenerates a coherent single exciton state. The second laser is tuned to biexciton resonances. By scanning the energy of the second laser for various polarizations of the two lasers, while monitoring the emission from the biexciton and exciton spectral lines, we map the biexciton photoluminescence excitation spectra. The resonances rich spectra of the second photon absorption are analyzed and fully understood in terms of a many carrier theoretical model which takes into account the direct and exchange Coulomb interactions between the quantum confined carriers.

  9. Holographic 3D multi-spot two-photon excitation for fast optical stimulation in brain

    Science.gov (United States)

    Takiguchi, Yu; Toyoda, Haruyoshi

    2017-04-01

    We report here a holographic high speed accessing microscope of sensory-driven synaptic activity across all inputs to single living neurons in the context of the intact cerebral cortex. This system is based on holographic multiple beam generation with spatial light modulator, we have demonstrated performance of the holographic excitation efficiency in several in vitro prototype system. 3D weighted iterative Fourier Transform method using the Ewald sphere in consideration of calculation speed has been adopted; multiple locations can be patterned in 3D with single hologram. Standard deviation of intensities of spots are still large due to the aberration of the system and/or hologram calculation, we successfully excited multiple locations of neurons in living mouse brain to monitor the calcium signals.

  10. Two-photon excitation of porphyrin-functionalized porous silicon nanoparticles for photodynamic therapy.

    Science.gov (United States)

    Secret, Emilie; Maynadier, Marie; Gallud, Audrey; Chaix, Arnaud; Bouffard, Elise; Gary-Bobo, Magali; Marcotte, Nathalie; Mongin, Olivier; El Cheikh, Khaled; Hugues, Vincent; Auffan, Mélanie; Frochot, Céline; Morère, Alain; Maillard, Philippe; Blanchard-Desce, Mireille; Sailor, Michael J; Garcia, Marcel; Durand, Jean-Olivier; Cunin, Frédérique

    2014-12-01

    Porous silicon nanoparticles (pSiNPs) act as a sensitizer for the 2-photon excitation of a pendant porphyrin using NIR laser light, for imaging and photodynamic therapy. Mannose-functionalized pSiNPs can be vectorized to MCF-7 human breast cancer cells through a mannose receptor-mediated endocytosis mechanism to provide a 3-fold enhancement of the 2-photon PDT effect.

  11. A Reversible DNA Logic Gate Platform Operated by One- and Two-Photon Excitations.

    Science.gov (United States)

    Tam, Dick Yan; Dai, Ziwen; Chan, Miu Shan; Liu, Ling Sum; Cheung, Man Ching; Bolze, Frederic; Tin, Chung; Lo, Pik Kwan

    2016-01-04

    We demonstrate the use of two different wavelength ranges of excitation light as inputs to remotely trigger the responses of the self-assembled DNA devices (D-OR). As an important feature of this device, the dependence of the readout fluorescent signals on the two external inputs, UV excitation for 1 min and/or near infrared irradiation (NIR) at 800 nm fs laser pulses, can mimic function of signal communication in OR logic gates. Their operations could be reset easily to its initial state. Furthermore, these DNA devices exhibit efficient cellular uptake, low cytotoxicity, and high bio-stability in different cell lines. They are considered as the first example of a photo-responsive DNA logic gate system, as well as a biocompatible, multi-wavelength excited system in response to UV and NIR. This is an important step to explore the concept of photo-responsive DNA-based systems as versatile tools in DNA computing, display devices, optical communication, and biology. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Polychromophoric metal complexes for generating the bioregulatory agent nitric oxide by single- and two-photon excitation.

    Science.gov (United States)

    Ford, Peter C

    2008-02-01

    In order to deliver a bioactive agent to a physiological location, it is important to be able to regulate precisely the location and the dosage. Such exquisite control can easily be envisioned for a photochemical drug that is active toward release of the desired bioactive agent upon irradiation of a specific tissue site. These materials should be thermally stable but reactive under excitation at visible (vis) or near-infrared (NIR) wavelengths where tissue transmission is optimal. Two photon excitation (TPE) is of special interest, since the use of focused laser pulses to activate release could provide 3D spatial control in therapeutic applications. This Account describes the preparation and photochemistry of a series of transition metal complexes designed to release the simple bioregulatory compound nitric oxide upon vis or NIR excitation. In order to enhance the light gathering capability of such compounds, we have attached chromophores with high single- or two-photon absorption cross sections to several photochemical NO precursors. For example, the iron nitrosyl clusters Fe2(mu-SR)2(NO)4 (Roussin's red esters) have been prepared with various chromophores as pendant groups, an example being the protoporphyrin XI derivative illustrated here. Direct excitation into the vis absorbing Q bands of the porphyrin leads to enhanced rates of NO generation from the Fe/S/NO cluster owing to the larger rate of light absorption by that antenna. Furthermore, femtosecond pulsed laser NIR excitation of the same compound at 810 nm (a spectral region where no absorption bands are apparent) leads to weak emission at approximately 630 nm and generation of NO, both effects providing evidence of a TPE mechanism. Roussin's red esters with other chromophores described here are even more effective for TPE-stimulated NO release. Another photochemical NO precursor discussed is the Cr(III) complex trans-Cr(L)(ONO)2(+) where L is a cyclic tetraamine such as cyclam. When L includes a

  13. MULTIPHOTON MICROSCOPIC IMAGING OF MOUSE INTESTINAL MUCOSA BASED ON TWO-PHOTON EXCITED FLUORESCENCE AND SECOND HARMONIC GENERATION

    Directory of Open Access Journals (Sweden)

    REN'AN XU

    2013-01-01

    Full Text Available Multiphoton microscopy (MPM, based on two-photon excited fluorescence and second harmonic generation, enables direct noninvasive visualization of tissue architecture and cell morphology in live tissues without the administration of exogenous contrast agents. In this paper, we used MPM to image the microstructures of the mucosa in fresh, unfixed, and unstained intestinal tissue of mouse. The morphology and distribution of the main components in mucosa layer such as columnar cells, goblet cells, intestinal glands, and a little collagen fibers were clearly observed in MPM images, and then compared with standard H&E images from paired specimens. Our results indicate that MPM combined with endoscopy and miniaturization probes has the potential application in the clinical diagnosis and in vivo monitoring of early intestinal cancer.

  14. MRT letter: Two-photon excitation-based 2pi light-sheet system for nano-lithography.

    Science.gov (United States)

    Mohan, Kavya; Mondal, Partha Pratim

    2015-01-01

    We propose two-photon excitation-based light-sheet technique for nano-lithography. The system consists of 2π-configured cylindrical lens system with a common geometrical focus. Upon superposition, the phase-matched counter-propagating light-sheets result in the generation of identical and equi spaced nano-bump pattern. Study shows a feature size of as small as few tens of nanometers with a inter-bump distance of few hundred nanometers. This technique overcomes some of the limitations of existing nano-lithography techniques, thereby, may pave the way for mass-production of nano-structures. Potential applications can also be found in optical microscopy, plasmonics, and nano-electronics. © 2014 Wiley Periodicals, Inc.

  15. Two-photon excited surface plasmon enhanced energy transfer between DAPI and gold nanoparticles: Opportunities in intra-cellular imaging and sensing

    Science.gov (United States)

    Zhang, Yinan; Birch, David J. S.; Chen, Yu

    2011-09-01

    We have demonstrated energy transfer between 4'-6-Diamidino-2-phenylindole (DAPI), a commonly used DNA label, and gold nanoparticles under two-photon excitation in solution using fluorescence lifetime imaging microscopy (FLIM). With comparable size and concentration, gold nanorods (GNRs) are shown to provide more efficient energy transfer than gold nanospheres (GNSs). We attribute this transfer enhancement effect to the longitudinal surface plasmon mode of GNRs overlapping with the excitation wavelength. Energy transfer under two-photon excitation between GNRs and DAPI has also been observed in cell culture and found to be in accord with the solution phase results.

  16. ARTICLES: A Surface Femtosecond Two-Photon Photoemission Spectrometer for Excited Electron Dynamics and Time-Dependent Photochemical Kinetics

    Science.gov (United States)

    Ren, Ze-feng; Zhou, Chuan-yao; Ma, Zhi-bo; Xiao, Chun-lei; Mao, Xin-chun; Dai, Dong-xu; LaRue, Jerry; Cooper, Russell; Wodtke, Alec M.; Yang, Xue-ming

    2010-06-01

    A surface femtosecond two-photon photoemission (2PPE) spectrometer devoted to the study of ultrafast excited electron dynamics and photochemical kinetics on metal and metal oxide surfaces has been constructed. Low energy photoelectrons are measured using a hemispherical electron energy analyzer with an imaging detector that allows us to detect the energy and the angular distributions of the photoelectrons simultaneously. A Mach-Zehnder interferometer was built for the time-resolved 2PPE (TR-2PPE) measurement to study ultrafast surface excited electron dynamics, which was demonstrated on the Cu(111) surface. A scheme for measuring time-dependent 2PPE (TD-2PPE) spectra has also been developed for studies of surface photochemistry. This technique has been applied to a preliminary study on the photochemical kinetics on ethanol/TiO2(110). We have also shown that the ultrafast dynamics of photoinduced surface excited resonances can be investigated in a reliable way by combining the TR-2PPE and TD-2PPE techniques.

  17. Two-photon excitation fluorescence imaging of the living juxtaglomerular apparatus.

    Science.gov (United States)

    Peti-Peterdi, János; Morishima, Shigeru; Bell, P Darwin; Okada, Yasunobu

    2002-07-01

    Recently, multiphoton excitation fluorescence microscopy has been developed that offers important advantages over confocal imaging, particularly for in vivo visualization of thick tissue samples. We used this state-of-the-art technique to capture high-quality images and study the function of otherwise inaccessible cell types and complex cell structures of the juxtaglomerular apparatus (JGA) in living preparations of the kidney. This structure has multiple cell types that exhibit a complex array of functions, which regulate the process of filtrate formation and renal hemodynamics. We report, for the first time, on high-resolution three-dimensional morphology and Z-sectioning through isolated, perfused kidney glomeruli, tubules, and JGA. Time-series images show how alterations in tubular fluid composition cause striking changes in single-cell volume of the unique macula densa tubular epithelium in situ and how they also affect glomerular filtration through alterations in associated structures within the JGA. In addition, calcium imaging of the glomerulus and JGA demonstrates the utility of this system in capturing the complexity of events and effects that are exerted by the specific hypertensive autacoid angiotensin II. This imaging approach to the study of isolated, perfused live tissue with multiphoton microscopy may be applied to other biological systems in which multiple cell types form a functionally integrated syncytium.

  18. Volumetric label-free imaging and 3D reconstruction of mammalian cochlea based on two-photon excitation fluorescence microscopy

    Science.gov (United States)

    Zhang, Xianzeng; Geng, Yang; Ye, Qing; Zhan, Zhenlin; Xie, Shusen

    2013-11-01

    The visualization of the delicate structure and spatial relationship of intracochlear sensory cells has relied on the laborious procedures of tissue excision, fixation, sectioning and staining for light and electron microscopy. Confocal microscopy is advantageous for its high resolution and deep penetration depth, yet disadvantageous due to the necessity of exogenous labeling. In this study, we present the volumetric imaging of rat cochlea without exogenous dyes using a near-infrared femtosecond laser as the excitation mechanism and endogenous two-photon excitation fluorescence (TPEF) as the contrast mechanism. We find that TPEF exhibits strong contrast, allowing cellular and even subcellular resolution imaging of the cochlea, differentiating cell types, visualizing delicate structures and the radial nerve fiber. Our results further demonstrate that 3D reconstruction rendered with z-stacks of optical sections enables better revealment of fine structures and spatial relationships, and easily performed morphometric analysis. The TPEF-based optical biopsy technique provides great potential for new and sensitive diagnostic tools for hearing loss or hearing disorders, especially when combined with fiber-based microendoscopy.

  19. Two-photon excitation spectroscopy of Cr3+:K2NaScF6 elpasolite: I. Experimental aspects

    Science.gov (United States)

    Wein, G. R.; Hamilton, D. S.; Sliwczuk, U.; Rinzler, A. G.; Bartram, R. H.

    2001-03-01

    Two-photon excitation experiments were performed to improve understanding of electron-lattice coupling and its effects on intra-3d3 transitions. Cr3+ occupies a scandium octahedral site in K2NaScF6. The transitions studied were 4A2g→ 4T2g and 4A2g→ 4T1ag. Complete spectra were recorded at a temperature of 10 K with the polarization vector or crystallographic direction. The two bands exhibit different polarization anisotropies and phonon couplings. The electric-dipole-forbidden 4A2g→ 4T2g band appears to be built on an eg-mode false origin and contains Fano antiresonances. This broad transition band lacks a zero-phonon line or any other sharp structure. The 4A2g→ 4T1ag transition zero-phonon line is evident and shows a 163 cm-1 low-temperature phase-transition-induced splitting. It also contains an extended progression of 35 phonon peaks corresponding to a lattice mode with phonon energy 106 cm-1, and a second progression with phonon energy 310 cm-1. The very asymmetric phonon side band displays a polarization anisotropy that differs from that of the zero-phonon line. To facilitate analysis of the data, measurements of low-temperature 4T2g→ 4A2g emission spectra with one-photon excitation are also reported and interpreted in the present paper.

  20. Fs-transient absorption and fluorescence upconversion after two- photon excitation of carotenoids in solution and in LHC II

    CERN Document Server

    Wall, P J; Fleming, G R

    2000-01-01

    With time resolved two-photon techniques we determined the lifetime and two-photon spectrum of the forbidden S/sub 1/ state of beta - carotene (9+or-0.2 ps), lutein (15+or-0.5 ps) and the energy transferring carotenoids in LHC II (250+or-50 fs). (7 refs).

  1. Photochemical Modulation of Ras-Mediated Signal Transduction using Caged Farnesyltransferase Inhibitors: Activation via One- and Two-Photon Excitation

    Science.gov (United States)

    Abate-Pella, Daniel; Zeliadt, Nicholette A.; Ochocki, Joshua D.; Warmka, Janel K.; Dore, Timothy M.; Blank, David A.; Wattenberg, Elizabeth V.; Distefano, Mark D.

    2012-01-01

    The creation of caged molecules involves the attachment of protecting groups to biologically active compounds such as ligands, substrates, and drugs that can be removed under specific conditions. Photoremovable caging groups are the most common due to their ability to be removed with high spatial and temporal resolution. Here, the synthesis and photochemistry of a caged inhibitor of protein farnesyltransferase, Bhc-FTI, is described. The inhibitor was caged by alkylation of a critical thiol functional group with a Bhc moiety; while Bhc is well established as a protecting group for carboxylates and phosphates, it has not been extensively used to cage sulfhydryls. The resulting caged molecule, Bhc-FTI, can be photolyzed with UV light to release the inhibitor (FTI) that prevents Ras farnesylation, Ras membrane localization and downstream signaling. Finally, it is shown that Bhc-FTI can be uncaged by two-photon excitation to produce FTI at levels sufficient to inhibit Ras localization and alter cell morphology. Given the widespread involvement of Ras proteins in signal transduction pathways, this caged inhibitor should be useful in a plethora of studies. PMID:22492666

  2. Photochemical modulation of Ras-mediated signal transduction using caged farnesyltransferase inhibitors: activation by one- and two-photon excitation.

    Science.gov (United States)

    Abate-Pella, Daniel; Zeliadt, Nicholette A; Ochocki, Joshua D; Warmka, Janel K; Dore, Timothy M; Blank, David A; Wattenberg, Elizabeth V; Distefano, Mark D

    2012-05-07

    The creation of caged molecules involves the attachment of protecting groups to biologically active compounds such as ligands, substrates and drugs that can be removed under specific conditions. Photoremovable caging groups are the most common due to their ability to be removed with high spatial and temporal resolution. Here, the synthesis and photochemistry of a caged inhibitor of protein farnesyltransferase is described. The inhibitor, FTI, was caged by alkylation of a critical thiol group with a bromohydroxycoumarin (Bhc) moiety. While Bhc is well established as a protecting group for carboxylates and phosphates, it has not been extensively used to cage sulfhydryl groups. The resulting caged molecule, Bhc-FTI, can be photolyzed with UV light to release the inhibitor that prevents Ras farnesylation, Ras membrane localization and downstream signaling. Finally, it is shown that Bhc-FTI can be uncaged by two-photon excitation to produce FTI at levels sufficient to inhibit Ras localization and alter cell morphology. Given the widespread involvement of Ras proteins in signal transduction pathways, this caged inhibitor should be useful in a plethora of studies.

  3. Stratum corneum lipid organization as observed by atomic force, confocal and two-photon excitation fluorescence microscopy.

    Science.gov (United States)

    Norlén, L; Plasencia, I; Bagatolli, L

    2008-12-01

    Skin moisturization is largely a function of stratum corneum barrier capacity, which in turn is a function of the physical state and structural organization of the stratum corneum extracellular lipid matrix [J. Invest. Dermatol.18, 433 (1952); AIChE J. 21, 985 (1975); Acta Derm. Venereol.74, 1 (1994); J. Invest. Dermatol.117, 830 (2001)]. Three unsolved key questions with respect to this lipid matrix' structural organization [Acta Derm. Venereol.74, 1 (1994); J. Invest. Dermatol.117, 830 (2001); J. Invest. Dermatol.118, 897 (2002); J. Invest. Dermatol.118, 899 (2002)] are: i) whether the lipid matrix is constituted by a single-gel phase or by co-existing solid (crystalline or gel) domains, ii) whether a separate fluid (liquid crystalline) phase is present and iii) whether the local pH has a direct effect on the lipid matrix' phase behaviour. Using an array of complementary visual-related biophysical techniques (e.g. atomic force microscopy and confocal/two-photon excitation fluorescence microscopy), it was recently shown that reconstituted membranes composed of extracted decontaminated human stratum corneum lipids do not form a fluid phase, but exclusively a single-gel phase that segregates into co-existing microscopic domains below pH 6 [Biophys. J.93, 3142 (2007)]. It was further shown that the role of cholesterol is related to dispersion of ceramide-enriched domains. This effect is counteracted by the presence of free fatty acids, which mix with skin ceramides but not with cholesterol.

  4. Endogenous Two-Photon Excited Fluorescence Provides Label-Free Visualization of the Inflammatory Response in the Rodent Spinal Cord

    Directory of Open Access Journals (Sweden)

    Ortrud Uckermann

    2015-01-01

    Full Text Available Activation of CNS resident microglia and invasion of external macrophages plays a central role in spinal cord injuries and diseases. Multiphoton microscopy based on intrinsic tissue properties offers the possibility of label-free imaging and has the potential to be applied in vivo. In this work, we analyzed cellular structures displaying endogenous two-photon excited fluorescence (TPEF in the pathologic spinal cord. It was compared qualitatively and quantitatively to Iba1 and CD68 immunohistochemical staining in two models: rat spinal cord injury and mouse encephalomyelitis. The extent of tissue damage was retrieved by coherent anti-Stokes Raman scattering (CARS and second harmonic generation imaging. The pattern of CD68-positive cells representing postinjury activated microglia/macrophages was colocalized to the TPEF signal. Iba1-positive microglia were found in areas lacking any TPEF signal. In peripheral areas of inflammation, we found similar numbers of CD68-positive microglia/macrophages and TPEF-positive structures while the number of Iba1-positive cells was significantly higher. Therefore, we conclude that multiphoton imaging of unstained spinal cord tissue enables retrieving the extent of microglia activation by acquisition of endogenous TPEF. Future application of this technique in vivo will enable monitoring inflammatory responses of the nervous system allowing new insights into degenerative and regenerative processes.

  5. Endogenous Two-Photon Excited Fluorescence Provides Label-Free Visualization of the Inflammatory Response in the Rodent Spinal Cord

    Science.gov (United States)

    Uckermann, Ortrud; Galli, Roberta; Beiermeister, Rudolf; Sitoci-Ficici, Kerim-Hakan; Later, Robert; Leipnitz, Elke; Neuwirth, Ales; Chavakis, Triantafyllos; Koch, Edmund; Schackert, Gabriele; Steiner, Gerald; Kirsch, Matthias

    2015-01-01

    Activation of CNS resident microglia and invasion of external macrophages plays a central role in spinal cord injuries and diseases. Multiphoton microscopy based on intrinsic tissue properties offers the possibility of label-free imaging and has the potential to be applied in vivo. In this work, we analyzed cellular structures displaying endogenous two-photon excited fluorescence (TPEF) in the pathologic spinal cord. It was compared qualitatively and quantitatively to Iba1 and CD68 immunohistochemical staining in two models: rat spinal cord injury and mouse encephalomyelitis. The extent of tissue damage was retrieved by coherent anti-Stokes Raman scattering (CARS) and second harmonic generation imaging. The pattern of CD68-positive cells representing postinjury activated microglia/macrophages was colocalized to the TPEF signal. Iba1-positive microglia were found in areas lacking any TPEF signal. In peripheral areas of inflammation, we found similar numbers of CD68-positive microglia/macrophages and TPEF-positive structures while the number of Iba1-positive cells was significantly higher. Therefore, we conclude that multiphoton imaging of unstained spinal cord tissue enables retrieving the extent of microglia activation by acquisition of endogenous TPEF. Future application of this technique in vivo will enable monitoring inflammatory responses of the nervous system allowing new insights into degenerative and regenerative processes. PMID:26355949

  6. Radiative electronic energy transfer-time studies of naphthalene-biacetyl system by one and two-photon excitation, and optical antenna mechanism.

    Science.gov (United States)

    Bayrakceken, Fuat

    2005-04-01

    In principle, the optical energy absorbed by a complex molecule raises that molecule to one of its excited states, and afterwards this excitation energy decays through the relaxation channels. Initially, electronically excited naphthalene emits photons and these emitted photons are absorbed by the acceptor molecule biacetyl, then excited biacetyl fluoresces. In this investigation radiative energy transfer-time is measured in cyclohexane by one and two-photon excitations. The UV-vis spectrum of biacetyl vapor at room temperature conditions was broad and structureless.

  7. Fluorescent detection and imaging of Hg{sup 2+} using a novel phenanthroline derivative based single- and two-photon excitation

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Xian, E-mail: zhangx@qlu.edu.cn; Li, Long-long; Liu, Ying-kai

    2016-02-01

    A novel phenanthroline derivative, 4-[4-(N-methyl)styrene]-imidazo[4,5-f][1,10]phenanthroline-benzene iodated salt (MSIPBI), was synthesized, and the linear absorption and fluorescent spectra of MSIPBI in different solvents were investigated. The photophysical properties in unbound and in ligand–metal complexes were evaluated by UV absorption and one- and two-photon fluorescent spectra, and the quantum yields, two-photon active cross-sections and the binding constant of dye–metal were calculated. The results indicated that MSIPBI has a large Stokes shift (more than 167 nm), and the dye was selective and sensitive for the detection of Hg{sup 2+} with a two-photon active cross-section of 55.5 GM in tris–HCl buffer solution at 800 nm. Furthermore, the results of the fluorescence microscopy imaging indicated that MSIPBI is an efficient fluorescent probe for the detection of Hg{sup 2+} in living cells by one- and two-photon excitation. Moreover, the experiments of determination Hg{sup 2+} in river water and tap water were finished. - Highlights: • A novel phenanthroline derivative (MSIPBI) has been synthesized. • The dye of MSIPBI was selective and sensitive to detect Hg{sup 2+}. • MSIPBI has a large Stokes shift (≥ 167 nm). • Hg{sup 2+} in living cells was successfully imaged by one- and two-photon excitation.

  8. Two-photon excitation of surface plasmon and the period-increasing effect of low spatial frequency ripples on a GaP crystal in air/water

    Science.gov (United States)

    Liu, Jukun; Jia, Tianqing; Zhao, Hongwei; Huang, Yaoqing

    2016-11-01

    We report the period-increasing effect of low spatial frequency ripples on a GaP crystal irradiated by 1 kHz, 50 fs, 800 nm femtosecond laser pulses. Massive free electrons are excited by a two-photon absorption process and surface plasmon is excited. The Drude model is used to estimate the changing of the dielectric constant of the GaP crystal. The period-increasing effects of low spatial frequency laser-induced ripples are theoretically predicted in air/water, and the experimental results agree well. The experimental and theoretical results indicate that surface plasmon excited by two-photon absorption plays a key role in the formation of low spatial frequency ripples.

  9. Photon Emission and Reabsorption Processes in CH3NH3PbBr3 Single Crystals Revealed by Time-Resolved Two-Photon-Excitation Photoluminescence Microscopy

    Science.gov (United States)

    Yamada, Takumi; Yamada, Yasuhiro; Nakaike, Yumi; Wakamiya, Atsushi; Kanemitsu, Yoshihiko

    2017-01-01

    The dynamical processes of radiative recombination of photocarriers and reabsorption of emitted photons in CH3NH3PbBr3 single crystals are studied using time-resolved two-photon-excitation photoluminescence (PL) microscopy. We find that the PL spectrum and its decay dynamics depend on the excitation-depth profile. As the excitation depth increases, the PL spectrum becomes asymmetric, the peak energy redshifts, and the PL decay time becomes longer. These observations can be well explained by a simple model including photon recycling (photon emission and reabsorption) in thick samples with strong band-to-band transitions and high radiative recombination efficiencies.

  10. The translated conceptual survey of physics / stablization of the focal plane in two photon excitation fluorescence microscopy

    Science.gov (United States)

    Wada, Asma

    As a reflection of my career to be an effective college physics teacher, my thesis is in two parts. The first is in education research, the focus of this part is to have a tool to evaluate pedagogies I have learned at the school and plan to apply in my classrooms back home. Consequently, this resulted in the development of the translated conceptual survey of physics ( TCSP). (TCSP) was designed by combining some questions from the Force Conceptual Inventory (FCI), and the Conceptual Survey of Electricity and Magnetism (CSEM) to assess student's understanding of basic concepts of Newtonian mechanics and electricity and magnetism in introductory physics. The idea of developing this questionnaire is to use it in classrooms back home as a part of a long term objective to implement what has been realized in the area of education research to improve the quality of teaching physics there. The survey was initially written in English, validated with interviews with native English speakers, translated into Arabic, and then validated via an interview with a native Arabic speaker. We then administered the survey to two different English-speaking intro physics courses and analyzed the results for consistency. The objective of the second part in my thesis is to expand my knowledge in an area of physics that I have interest in, and getting involved in a scientific research to develop skills I need as a teacher. My research is in optical physics, in particular, I am working on one of the challenges in implementing two photon excitation luorescence (TPEF) microscopy in imaging living systems. (TPEF) microscopy has been shown to be an invaluable tool for investigating biological structure and function in living organisms. The utility of (TPEF) imaging for this application arises from several important factors including it's ability to image deep within tissue, and to do so without harming the organism. Both of these advantages arise from the fact that (TPEF) imaging is done with

  11. Two-photon excited endogenous fluorescence for label-free in vivo imaging ingestion of disease-causing bacteria by human leukocytes

    Science.gov (United States)

    Zeng, Yan; Yan, Bo; Sun, Qiqi; Teh, Seng Khoon; Zhang, Wei; Wen, Zilong; Qu, Jianan Y.

    2013-02-01

    Real time and in vivo monitoring leukocyte behavior provides unique information to understand the physiological and pathological process of infection. In this study, we demonstrate that two-photon excited reduced nicotinamide adenine dinucleotide (NADH) fluorescence provides imaging contrast to distinguish granulocyte and agranulocyte. By using spectral and time-resolved NADH fluorescence, we study the immune response of human neutrophils against bacterial infection (Escherichia coli). The two-photon excited NADH fluorescence images clearly review the morphological changes from resting neutrophils (round shape) to activated neutrophils (ruffle shape) during phagocytosis. The free-tobound NADH ratio of neutrophils decreases after ingesting disease-causing pathogen: Escherichia coli. This finding may provide a new optical tool to investigate inflammatory processes by using NADH fluorescence in vivo.

  12. Two-photon excitation of the 2Π(4p)-X2Π(3p) transition of AlAr

    Science.gov (United States)

    Mascaritolo, Kyle J.; Antonov, Ivan O.; Heaven, Michael C.

    2014-03-01

    The 2Π(4p)-X2Π(3p) band system of AlAr has been observed using two-photon excitation. The spectrum consists of a short progression of doublet bands, with spin-orbit intervals that are close to that of Al(4p). Potential energy curve fitting yielded a bond dissociation energy for 2Π(4p) of De = 495(5) cm-1 and an approximate bond length of Re = 3.33(4) Å.

  13. Nonlinear spectral imaging of human hypertrophic scar based on two-photon excited fluorescence and second-harmonic generation.

    Science.gov (United States)

    Chen, G; Chen, J; Zhuo, S; Xiong, S; Zeng, H; Jiang, X; Chen, R; Xie, S

    2009-07-01

    A noninvasive method using microscopy and spectroscopy for analysing the morphology of collagen and elastin and their biochemical variations in skin tissue will enable better understanding of the pathophysiology of hypertrophic scars and facilitate improved clinical management and treatment of this disease. To obtain simultaneously microscopic images and spectra of collagen and elastin fibres in ex vivo skin tissues (normal skin and hypertrophic scar) using a nonlinear spectral imaging method, and to compare the morphological structure and spectral characteristics of collagen and elastin fibres in hypertrophic scar tissues with those of normal skin, to determine whether this approach has potential for in vivo assessment of the pathophysiology of human hypertrophic scars and for monitoring treatment responses as well as for tracking the process of development of hypertrophic scars in clinic. Ex vivo human skin specimens obtained from six patients aged from 10 to 50 years old who were undergoing skin plastic surgery were examined. Five patients had hypertrophic scar lesions and one patient had no scar lesion before we obtained his skin specimen. A total of 30 tissue section samples of 30 mum thickness were analysed by the use of a nonlinear spectral imaging system consisting of a femtosecond excitation light source, a high-throughput scanning inverted microscope, and a spectral imaging detection system. The high-contrast and high-resolution second harmonic generation (SHG) images of collagen and two-photon excited fluorescence (TPEF) images of elastin fibres in hypertrophic scar tissues and normal skin were acquired using the extracting channel tool of the system. The emission spectra were analysed using the image-guided spectral analysis method. The depth-dependent decay constant of the SHG signal and the image texture characteristics of hypertrophic scar tissue and normal skin were used to quantitatively assess the amount, distribution and orientation of their

  14. Fluorescence Detection of H5N1 Virus Gene Sequences Based on Optical Tweezers with Two-Photon Excitation Using a Single Near Infrared Nanosecond Pulse Laser.

    Science.gov (United States)

    Li, Cheng-Yu; Cao, Di; Kang, Ya-Feng; Lin, Yi; Cui, Ran; Pang, Dai-Wen; Tang, Hong-Wu

    2016-04-19

    We present an analytical platform by combining near-infrared optical tweezers with two-photon excitation for fluorescence detection of H5N1 virus gene sequences. A heterogeneous enrichment strategy, which involved polystyrene (PS) microsphere and quantum dots (QDs), was adopted. The final hybrid-conjugate microspheres were prepared by a facile one-step hybridization procedure by using PS microspheres capturing target DNA and QDs tagging, respectively. Quantitative detection was achieved by the optical tweezers setup with a low-cost 1064 nm nanosecond pulse laser for both optical trapping and two-photon excitation for the same hybrid-conjugate microsphere. The detection limits for both neuraminidase (NA) gene sequences and hemagglutinin (HA) gene sequences are 16-19 pM with good selectivity for one-base mismatch, which is approximately 1 order of magnitude lower than the most existing fluorescence-based analysis method. Besides, because of the fact that only signal from the trapped particle is detected upon two-photon excitation, this approach showed extremely low background in fluorescence detection and was successfully applied to directly detect target DNA in human whole serum without any separation steps and the corresponding results are very close to that in buffer solution, indicating the strong anti-interference ability of this method. Therefore, it can be expected to be an emerging alternative for straightforward detecting target species in complex samples with a simple procedure and high-throughput.

  15. Evaluation of the contribution of the renal capsule and cortex to kidney autofluorescence intensity under ultraviolet excitation

    Energy Technology Data Exchange (ETDEWEB)

    Raman, R N; Pivetti, C D; Rubenchik, A M; Matthews, D L; Troppmann, C; Demos, S G

    2008-12-12

    The use of reduced nicotinamide adenine dinucleotide (NADH) fluorescence to gain metabolic information on kidneys in response to an alteration in oxygen availability has previously been experimentally demonstrated, but signal quantification has not to date been addressed. In this work the relative contribution to rat kidney autofluorescence of the capsule vs. cortex under ultraviolet excitation is determined from experimental results obtained using autofluorescence microscopy and a suitable mathematical model. The results allow for a quantitative assessment of the relative contribution of the signal originating in the metabolically active cortex as a function of capsule thickness for different wavelengths.

  16. Three-dimensional microfabrication of protein hydrogels via two-photon-excited thiol-vinyl ester photopolymerization

    OpenAIRE

    Qin, Xiao-Hua; Torgersen, Jan; Saf, Robert; Mühleder, Severin; Pucher, Niklas; Ligon, Clark; Holnthoner, Wolfgang; Redl, Heinz; Ovsianikov, Aleksandr; Stampfl, Jürgen; Liska, Robert

    2013-01-01

    Engineering three-dimensional (3D) hydrogels with well-defined architectures has become increasingly important for tissue engineering and basic research in biomaterials science. To fabricate 3D hydrogels with (sub)cellular-scale features, two-photon polymerization (2PP) shows great promise although the technique is limited by the selection of appropriate hydrogel precursors. In this study, we report the synthesis of gelatin hydrolysate vinyl esters (GH-VE) and its copolymerization with reduce...

  17. Two-photon cryomicroscope

    Science.gov (United States)

    Breunig, H. G.; Köhler, C.; König, K.

    2012-03-01

    We report on a new two-photon cryomicroscope which consist of a compact laser-scanning microscope combined with a motorized heating and freezing stage. Samples can be cooled down to -196 °C (77 K) and heated up to 600 °C (873 K) with adjustable heating/freezing rates between 0.01 K / min and 150 K / min. Two-photon imaging is realized by near infrared femtosecond-laser pulse excitation. The abilities of the two-photon cryomicroscope are illustrated in several measurements: imaging of fluorescent microspheres inside a piece of ice illustrates the feasibility of deep-microscopic imaging inside frozen sample. The temperature-dependent structural integrity of collagen is monitored by detection of second harmonic generation signals from porcine cornea. The measurements reveal also the dependence of the collagendenaturation temperature on hydration state of the cornea collagen. Furthermore, the potential of the two-photon cryomicroscope for optimization of freezing and thawing procedures as well as to evaluate the viability of frozen cells and tissue is discussed.

  18. Label-free distinguishing between neurons and glial cells based on two-photon excited fluorescence signal of neuron perinuclear granules

    Science.gov (United States)

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

    2016-08-01

    Neurons and glial cells are two critical cell types of brain tissue. Their accurate identification is important for the diagnosis of psychiatric disorders such as depression and schizophrenia. In this paper, distinguishing between neurons and glial cells by using the two-photon excited fluorescence (TPEF) signals of intracellular intrinsic sources was performed. TPEF microscopy combined with TUJ-1 and GFAP immunostaining and quantitative image analysis demonstrated that the perinuclear granules of neurons in the TPEF images of brain tissue and the primary cultured cortical cells were a unique characteristic of neurons compared to glial cells which can become a quantitative feature to distinguish neurons from glial cells. With the development of miniaturized TPEF microscope (‘two-photon fiberscopes’) imaging devices, TPEF microscopy can be developed into an effective diagnostic and monitoring tool for psychiatric disorders such as depression and schizophrenia.

  19. Second harmonic imaging of plants tissues and cell implosion using two-photon process in ZnO nanoparticles.

    Science.gov (United States)

    Urban, Ben E; Neogi, Purnima B; Butler, Sween J; Fujita, Yasuhisa; Neogi, Arup

    2012-03-01

    The optical properties of colloidal ZnO nanoparticle (NP) solutions, with size ranging from several nm to around 200 nm, have been tailored to have high optical nonlinearity for bioimaging with no auto-fluorescence above 750 nm and minimal auto-fluorescence below 750 nm. The high second harmonic conversion efficiency enables selective tissue imaging and cell tracking using tunable near-infrared femtosecond laser source ranging from 750-980 nm. For laser energies exceeding the two-photon energy of the bandgap of ZnO (half of 3.34 eV), the SHG signal greatly decreases and the two-photon emission becomes the dominant signal. The heat generated due to two-photon absorption within the ZnO NPs enable selective cell or localized tissue destruction using excitation wavelength ranging from 710-750 nm. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Effects of two-photon absorption on terahertz radiation generated by femtosecond-laser excited photoconductive antennas.

    Science.gov (United States)

    Lee, Chao-Kuei; Yang, Chan-Shan; Lin, Sung-Hui; Huang, Shiuan-Hua; Wada, Osamu; Pan, Ci-Ling

    2011-11-21

    Terahertz (THz) radiation can be generated more efficiently from a low-temperature-grown GaAs (LT-GaAs) photoconductive (PC) antenna by considering the two-photon absorption (TPA) induced photo-carrier in the photoconductor. A rate-equation-based approach using the Drude-Lorentz model taking into account the band-diagram of LT-GaAs is used for the theoretical analysis. The use of transform-limited pulses at the PC antenna is critical experimentally. Previously unnoticed THz pulse features and anomalously increasing THz radiation power rather than saturation were observed. These are in good agreement with the theoretical predictions. The interplay of intensity dependence and dynamics of generation of photoexcited carriers by single-photon absorption and TPA for THz emission is discussed.

  1. Kinetics and subcellular localization of 5-ALA-induced PpIX in DHL cells via two-photon excitation fluorescence microscopy.

    Science.gov (United States)

    Chen, Rong; Huang, Zufang; Chen, Guannan; Li, Yongzeng; Chen, Xianlian; Chen, Jianxin; Zeng, Haishan

    2008-04-01

    Two-photon excitation fluorescence (TPEF) microscopy was used to measure the 5-aminolevulinic acid (5-ALA)-induced PpIX fluorescence in follicular lymphoma DHL cells. Kinetics of 5-ALA-induced PpIX accumulation in DHL cells under various 5-ALA concentrations was studied. We found that during the course of continuous incubation with 5-ALA, the relationship between the DHL cell fluorescence signal and the incubation time showed a biphasic variation. Initially the PpIX signal increased with the incubation time and reached the maximal value at about 3 h, and then it decreased with time during the subsequent incubation period. By labeling the 5-ALA incubated DHL cells with different organelle-specific fluorescence probes: Rhodamine 123 (for mitochondria), DioC6(3) (for endoplasmic reticulum) and LysoTracker Green (for lysosomes) respectively, we found that 5-ALA-induced PpIX was primarily localized in endoplasmic reticulum and mitochondria; its concentration in the lysosome was much lower. The results suggested that 5-ALA could potentially be an effective photosensitizer in photodynamic purging of DHL cells. Two-photon excitation fluorescence microscope is a useful tool for studying 5-ALA-induced PpIX subcellular localization.

  2. Two-photon excited fluorescence from higher electronic states of chlorophylls in photosynthetic antenna complexes a new approach to detect strong excitonic chlorophyll a/b coupling

    CERN Document Server

    Leupold, D; Ehlert, J; Irrgang, K D; Renger, G; Lokstein, H

    2002-01-01

    Stepwise two-photon excitation of chlorophyll a and b in the higher plant main light-harvesting complex (LHC II) and the minor complex CP29 (as well as in organic solution) with 100-fs pulses in the Q/sub y/ region results in a weak blue fluorescence. The dependence of the spectral shape of the blue fluorescence on excitation wavelength offers a new approach to elucidate the long-standing problem of the origin of spectral "chlorophyll forms" in pigment-protein complexes, in particular the characterization of chlorophyll a/b-heterodimers. As a first result we present evidence for the existence of strong chlorophyll a/b-interactions (excitonically coupled transitions at 650 and 680 nm) in LHC II at ambient temperature. In comparison with LHC II, the experiments with CP29 provide further evidence that the lowest energy chlorophyll a transition (at ~680 nm) is not excitonically coupled to chlorophyll b. (22 refs).

  3. Two-photon excitation laser scanning microscopy of rabbit nasal septal cartilage following Nd:YAG-laser-mediated stress relaxation

    Science.gov (United States)

    Kim, Charlton C.; Wallace, Vincent P.; Coleno, Mariah L.; Dao, Xavier; Tromberg, Bruce J.; Wong, Brian J.

    2000-04-01

    Laser irradiation of hyaline cartilage result in stable shape changes due to temperature dependent stress relaxation. In this study, we determined the structural changes in chondrocytes within rabbit nasal septal cartilage tissue over a 12-day period using a two-photon laser scanning microscope (TPM) following Nd:YAG laser irradiation. During laser irradiation surface temperature, stress relaxation, and diffuse reflectance, were measured dynamically. Each specimen received one or two sequential laser exposures. The cartilage reached a peak surface temperature of about 61 degrees C during irradiation. Cartilage denatured in 50 percent EtOH was used as a positive control. TPM was performed to detect the fluorescence emission from the chondrocytes. Images of chondrocytes were obtained at depths up to 150 microns, immediately following laser exposure, and also following 12 days in culture. Few differences in the pattern or intensity of fluorescence was observed between controls and irradiated specimens imaged immediately following exposure, regardless of the number of laser pulses. However, following twelve days in tissue culture, the irradiated specimens increase, whereas the native tissue diminishes, in intensity and distribution of fluorescence in the cytoplasm. In contrast, the positive control shows only extracellular matrices and empty lacuna, feature consistent with cell membrane lysis.

  4. Two-photon absorption spectroscopy of stilbene and phenanthrene: Excited-state analysis and comparison with ethylene and toluene

    Science.gov (United States)

    de Wergifosse, Marc; Elles, Christopher G.; Krylov, Anna I.

    2017-05-01

    Two-photon absorption (2PA) spectra of several prototypical molecules (ethylene, toluene, trans- and cis-stilbene, and phenanthrene) are computed using the equation-of-motion coupled-cluster method with single and double substitutions. The states giving rise to the largest 2PA cross sections are analyzed in terms of their orbital character and symmetry-based selection rules. The brightest 2PA transitions correspond to Rydberg-like states from fully symmetric irreducible representations. Symmetry selection rules dictate that totally symmetric transitions typically have the largest 2PA cross sections for an orientationally averaged sample when there is no resonance enhancement via one-photon accessible intermediate states. Transition dipole arguments suggest that the strongest transitions also involve the most delocalized orbitals, including Rydberg states, for which the relative transition intensities can be rationalized in terms of atomic selection rules. Analysis of the 2PA transitions provides a foundation for predicting relative 2PA cross sections of conjugated molecules based on simple symmetry and molecular orbital arguments.

  5. Nonlinear spectral imaging of human normal skin, basal cell carcinoma and squamous cell carcinoma based on two-photon excited fluorescence and second-harmonic generation

    Science.gov (United States)

    Xiong, S. Y.; Yang, J. G.; Zhuang, J.

    2011-10-01

    In this work, we use nonlinear spectral imaging based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) for analyzing the morphology of collagen and elastin and their biochemical variations in basal cell carcinoma (BCC), squamous cell carcinoma (SCC) and normal skin tissue. It was found in this work that there existed apparent differences among BCC, SCC and normal skin in terms of their thickness of the keratin and epithelial layers, their size of elastic fibers, as well as their distribution and spectral characteristics of collagen. These differences can potentially be used to distinguish BCC and SCC from normal skin, and to discriminate between BCC and SCC, as well as to evaluate treatment responses.

  6. Effect of detergents on the physico-chemical properties of skin stratum corneum: A two-photon excitation fluorescence microscopy study

    DEFF Research Database (Denmark)

    Bloksgaard, Maria; Brewer, Jonathan R.; Pashkovski, Eugene

    2014-01-01

    performed using two-photon excitation fluorescence microscopy. Fluorescent images of fluorescence reporters sensitive to proton activity and hydration of SC were obtained in excised skin and examined in presence and absence of SCM and SDS detergents. RESULTS: Hydration of the intercellular lipid matrix......OBJECTIVE: Understanding the structural and dynamical features of skin is critical for advancing innovation in personal care and drug discovery. Synthetic detergent mixtures used in commercially available body wash products are thought to be less aggressive towards the skin barrier when compared...... to conventional detergents. The aim of this work is to comparatively characterize the effect of a mild synthetic cleanser mixture (SCM) and sodium dodecyl sulphate (SDS) on the hydration state of the intercellular lipid matrix and on proton activity of excised skin stratum corneum (SC). METHOD: Experiments were...

  7. Correction of depth-induced spherical aberration for deep observation using two-photon excitation fluorescence microscopy with spatial light modulator.

    Science.gov (United States)

    Matsumoto, Naoya; Inoue, Takashi; Matsumoto, Akiyuki; Okazaki, Shigetoshi

    2015-07-01

    We demonstrate fluorescence imaging with high fluorescence intensity and depth resolution in which depth-induced spherical aberration (SA) caused by refractive-index mismatch between the medium and biological sample is corrected. To reduce the impact of SA, we incorporate a spatial light modulator into a two-photon excitation fluorescence microscope. Consequently, when fluorescent beads in epoxy resin were observed with this method of SA correction, the fluorescence signal of the observed images was ∼27 times higher and extension in the direction of the optical axes was ∼6.5 times shorter at a depth of ∼890 μm. Thus, the proposed method increases the depth observable at high resolution. Further, our results show that the method improved the fluorescence intensity of images of the fluorescent beads and the structure of a biological sample.

  8. Comparison of nanosecond and picosecond excitation for interference-free two-photon laser-induced fluorescence detection of atomic hydrogen in flames.

    Science.gov (United States)

    Kulatilaka, Waruna D; Patterson, Brian D; Frank, Jonathan H; Settersten, Thomas B

    2008-09-10

    Two-photon laser-induced fluorescence (TP-LIF) line imaging of atomic hydrogen was investigated in a series of premixed CH4/O2/N2, H2/O2, and H2/O2/N2 flames using excitation with either picosecond or nanosecond pulsed lasers operating at 205 nm. Radial TP-LIF profiles were measured for a range of pulse fluences to determine the maximum interference-free signal levels and the corresponding picosecond and nanosecond laser fluences in each of 12 flames. For an interference-free measurement, the shape of the TP-LIF profile is independent of laser fluence. For larger fluences, distortions in the profile are attributed to photodissociation of H2O, CH3, and/or other combustion intermediates, and stimulated emission. In comparison with the nanosecond laser, excitation with the picosecond laser can effectively reduce the photolytic interference and produces approximately an order of magnitude larger interference-free signal in CH4/O2/N2 flames with equivalence ratios in the range of 0.5laser fluence in all flames, stimulated emission, occurring between the laser-excited level, H(n=3), and H(n=2), is the limiting factor for picosecond excitation in the flames with the highest H atom concentration. Nanosecond excitation is advantageous in the richest (Phi=1.64) CH4/O2/N2 flame and in H2/O2/N2 flames. The optimal excitation pulse width for interference-free H atom detection depends on the relative concentrations of hydrogen atoms and photolytic precursors, the flame temperature, and the laser path length within the flame.

  9. Two-photon excitation in chip electrophoresis enabling label-free fluorescence detection in non-UV transparent full-body polymer chips.

    Science.gov (United States)

    Geissler, David; Belder, Detlev

    2015-12-01

    One of the most commonly employed detection methods in microfluidic research is fluorescence detection, due to its ease of integration and excellent sensitivity. Many analytes though do not show luminescence when excited in the visible light spectrum, require suitable dyes. Deep-ultraviolet (UV) excitation (body polymer microfluidic devices. This was achieved by means of two-photon excitation in the visible range (λex = 532 nm). Issues associated with the low optical transmittance of plastics in the UV range were successfully circumvented in this way. The technique was investigated by application to microchip electrophoresis of small aromatic compounds. Various polymers, such as poly(methyl methacrylate), cyclic olefin polymer, and copolymer as well as poly(dimethylsiloxane) were investigated and compared with respect to achievable LOD and ruggedness against photodamage. To demonstrate the applicability of the technique, the method was also applied to the determination of serotonin and tryptamine in fruit samples. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Center for Fluorescence Spectroscopy: advanced studies of fluorescence dynamics, lifetime imaging, clinical sensing, two-photon excitation, and light quenching

    Science.gov (United States)

    Lakowicz, Joseph R.; Malak, Henryk M.; Gryczynski, Ignacy; Szmacinski, Henryk; Kusba, Jozef; Akkaya, Engin; Terpetschnig, Ewald A.; Johnson, Michael L.

    1994-08-01

    The Center for Fluorescence Spectroscopy (CFS) is a multi-user facility providing state of the art time-resolved fluorescence instrumentation and software for scientists, whose research can be enhanced by such experimental data. The CFS is a national center, supported by the National Center for Research Resources Division of the National Institutes of Health, and in part by the National Science Foundation. Both time-domain (TD) and frequency- domain (FD) measurements (10 MHz to 10 Ghz) are available, with a wide range of excitation and emission wavelengths (UV to NIR). The data can be used to recover distances and site-to-site diffusion in protein, interactions between macromolecules, accessibility of fluorophores to quenchers, and the dynamic properties of proteins, membranes and nucleic acids. Current software provides for analysis of multi-exponential intensity and anisotropy decays, lifetime distribution, distance distributions for independent observation of fluorescence donors and acceptors, transient effects in collisional quenching, phase-modulation spectra and time-resolved emission spectra. Most programs provide for global analysis of multiple data sets obtained under similar experimental conditions. Data can be analyzed on-site by connection with the CFS computers through the internet. During six years of operation we have established scientific collaborations with over 30 academic and industrial groups in the United States. These collaborations have resulted in 63 scientific papers.

  11. Label-free imaging of brain and brain tumor specimens with combined two-photon excited fluorescence and second harmonic generation microscopy

    Science.gov (United States)

    Jiang, Liwei; Wang, Xingfu; Wu, Zanyi; Du, Huiping; Wang, Shu; Li, Lianhuang; Fang, Na; Lin, Peihua; Chen, Jianxin; Kang, Dezhi; Zhuo, Shuangmu

    2017-10-01

    Label-free imaging techniques are gaining acceptance within the medical imaging field, including brain imaging, because they have the potential to be applied to intraoperative in situ identifications of pathological conditions. In this paper, we describe the use of two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) microscopy in combination for the label-free detection of brain and brain tumor specimens; gliomas. Two independently detecting channels were chosen to subsequently collect TPEF/SHG signals from the specimen to increase TPEF/SHG image contrasts. Our results indicate that the combined TPEF/SHG microscopic techniques can provide similar rat brain structural information and produce a similar resolution like conventional H&E staining in neuropathology; including meninges, cerebral cortex, white-matter structure corpus callosum, choroid plexus, hippocampus, striatum, and cerebellar cortex. It can simultaneously detect infiltrating human brain tumor cells, the extracellular matrix collagen fiber of connective stroma within brain vessels and collagen depostion in tumor microenvironments. The nuclear-to-cytoplasmic ratio and collagen content can be extracted as quantitative indicators for differentiating brain gliomas from healthy brain tissues. With the development of two-photon fiberscopes and microendoscope probes and their clinical applications, the combined TPEF and SHG microcopy may become an important multimodal, nonlinear optical imaging approach for real-time intraoperative histological diagnostics of residual brain tumors. These occur in various brain regions during ongoing surgeries through the method of simultaneously identifying tumor cells, and the change of tumor microenvironments, without the need for the removal biopsies and without the need for tissue labelling or fluorescent markers.

  12. Ratiometric two-photon excited photoluminescence of quantum dots triggered by near-infrared-light for real-time detection of nitric oxide release in situ

    Energy Technology Data Exchange (ETDEWEB)

    Jin, Hui [Shandong Sino-Japanese Center for Collaborative Research of Carbon Nanomaterials, Collaborative Innovation Center for Marine Biomass Fiber Materials and Textiles, College of Chemistry and Chemical Engineering, Laboratory of Fiber Materials and Modern Textile, The Growing Base for State Key Laboratory, Qingdao University, Shandong 266071 (China); Gui, Rijun, E-mail: guirijun@qdu.edu.cn [Shandong Sino-Japanese Center for Collaborative Research of Carbon Nanomaterials, Collaborative Innovation Center for Marine Biomass Fiber Materials and Textiles, College of Chemistry and Chemical Engineering, Laboratory of Fiber Materials and Modern Textile, The Growing Base for State Key Laboratory, Qingdao University, Shandong 266071 (China); Sun, Jie; Wang, Yanfeng [Institute of Materia Medica, Shandong Academy of Medical Sciences, Jinan 250062 (China)

    2016-05-30

    Probe-donor integrated nanocomposites were developed from conjugating silica-coated Mn{sup 2+}:ZnS quantum dots (QDs) with MoS{sub 2} QDs and photosensitive nitric oxide (NO) donors (Fe{sub 4}S{sub 3}(NO){sub 7}{sup −}, RBS). Under excitation with near-infrared (NIR) light at 808 nm, the Mn{sup 2+}:ZnS@SiO{sub 2}/MoS{sub 2}-RBS nanocomposites showed the dual-emissive two-photon excited photoluminescence (TPEPL) that induced RBS photolysis to release NO in situ. NO caused TPEPL quenching of Mn{sup 2+}:ZnS QDs, but it produced almost no impact on the TPEPL of MoS{sub 2} QDs. Hence, the nanocomposites were developed as a novel QDs-based ratiometric TPEPL probe for real-time detection of NO release in situ. The ratiometric TPEPL intensity is nearly linear (R{sup 2} = 0.9901) with NO concentration in the range of 0.01∼0.8 μM, which corresponds to the range of NO release time (0∼15 min). The detection limit was calculated to be approximately 4 nM of NO. Experimental results confirmed that this novel ratiometric TPEPL probe possessed high selectivity and sensitivity for the detection of NO against potential competitors, and especially showed high detection performance for NIR-light triggered NO release in tumor intracellular microenvironments. These results would promote the development of versatile probe-donor integrated systems, also providing a facile and efficient strategy to real-time detect the highly controllable drug release in situ, especially in physiological microenvironments. - Highlights: • Mn{sup 2+}:ZnS@SiO{sub 2}/MoS{sub 2}-RBS nanocomposites were developed as a novel ratiometric two-photon excited fluorescence probe. • This probe could conduct real-time detection of nitric oxide release in situ. • High feasibility of this probe was confirmed in tumor intracellular microenvironments.

  13. Correlating two-photon excited fluorescence imaging of breast cancer cellular redox state with seahorse flux analysis of normalized cellular oxygen consumption

    Science.gov (United States)

    Hou, Jue; Wright, Heather J.; Chan, Nicole; Tran, Richard; Razorenova, Olga V.; Potma, Eric O.; Tromberg, Bruce J.

    2016-06-01

    Two-photon excited fluorescence (TPEF) imaging of the cellular cofactors nicotinamide adenine dinucleotide and oxidized flavin adenine dinucleotide is widely used to measure cellular metabolism, both in normal and pathological cells and tissues. When dual-wavelength excitation is used, ratiometric TPEF imaging of the intrinsic cofactor fluorescence provides a metabolic index of cells-the "optical redox ratio" (ORR). With increased interest in understanding and controlling cellular metabolism in cancer, there is a need to evaluate the performance of ORR in malignant cells. We compare TPEF metabolic imaging with seahorse flux analysis of cellular oxygen consumption in two different breast cancer cell lines (MCF-7 and MDA-MB-231). We monitor metabolic index in living cells under both normal culture conditions and, for MCF-7, in response to cell respiration inhibitors and uncouplers. We observe a significant correlation between the TPEF-derived ORR and the flux analyzer measurements (R=0.7901, p<0.001). Our results confirm that the ORR is a valid dynamic index of cell metabolism under a range of oxygen consumption conditions relevant for cancer imaging.

  14. Two-photon-excited fluorescence (TPEF) and fluorescence lifetime imaging (FLIM) with sub-nanosecond pulses and a high analog bandwidth signal detection

    Science.gov (United States)

    Eibl, Matthias; Karpf, Sebastian; Hakert, Hubertus; Weng, Daniel; Huber, Robert

    2017-02-01

    Two-photon excited fluorescence (TPEF) microscopy and fluorescence lifetime imaging (FLIM) are powerful imaging techniques in bio-molecular science. The need for elaborate light sources for TPEF and speed limitations for FLIM, however, hinder an even wider application. We present a way to overcome this limitations by combining a robust and inexpensive fiber laser for nonlinear excitation with a fast analog digitization method for rapid FLIM imaging. The applied sub nanosecond pulsed laser source is synchronized to a high analog bandwidth signal detection for single shot TPEF- and single shot FLIM imaging. The actively modulated pulses at 1064nm from the fiber laser are adjustable from 50ps to 5ns with kW of peak power. At a typically applied pulse lengths and repetition rates, the duty cycle is comparable to typically used femtosecond pulses and thus the peak power is also comparable at same cw-power. Hence, both types of excitation should yield the same number of fluorescence photons per time on average when used for TPEF imaging. However, in the 100ps configuration, a thousand times more fluorescence photons are generated per pulse. In this paper, we now show that the higher number of fluorescence photons per pulse combined with a high analog bandwidth detection makes it possible to not only use a single pulse per pixel for TPEF imaging but also to resolve the exponential time decay for FLIM. To evaluate the performance of our system, we acquired FLIM images of a Convallaria sample with pixel rates of 1 MHz where the lifetime information is directly measured with a fast real time digitizer. With the presented results, we show that longer pulses in the many-10ps to nanosecond regime can be readily applied for TPEF imaging and enable new imaging modalities like single pulse FLIM.

  15. Two-photon physics

    Energy Technology Data Exchange (ETDEWEB)

    Bardeen, W.A.

    1981-10-01

    A new experimental frontier has recently been opened to the study of two photon processes. The first results of many aspects of these reactions are being presented at this conference. In contrast, the theoretical development of research ito two photon processes has a much longer history. This talk reviews the many different theoretical ideas which provide a detailed framework for our understanding of two photon processes.

  16. Combined nonlinear laser imaging (two-photon excitation fluorescence, second and third-harmonic generation, and fluorescence lifetime imaging microscopies) in ovarian tumors

    Science.gov (United States)

    Adur, J.; Pelegati, V. B.; de Thomaz, A. A.; Bottcher-Luiz, F.; Andrade, L. A. L. A.; Almeida, D. B.; Carvalho, H. F.; Cesar, C. L.

    2012-03-01

    We applied Two-photon Excited Fluorescence (TPEF), Second/Third Harmonic Generation (SHG and THG) and Fluorescence Lifetime Imaging (FLIM) Non Linear Optics (NLO) Laser-Scanning Microscopy within the same imaging platform to evaluate their use as a diagnostic tool in ovarian tumors. We assess of applicability of this multimodal approach to perform a pathological evaluation of serous and mucinous tumors in human samples. The combination of TPEF-SHG-THG imaging provided complementary information about the interface epithelium/stromal, such as the transformation of epithelium surface (THG) and the overall fibrillar tissue architecture (SHG). The fact that H&E staining is the standard method used in clinical pathology and that the stored samples are usually fixed makes it important a re-evaluation of these samples with NLO microscopy to compare new results with a library of already existing samples. FLIM, however, depends on the chemical environment around the fluorophors that was completely changed after fixation; therefore it only makes sense in unstained samples. Our FLIM results in unstained samples demonstrate that it is possible to discriminate healthy epithelia from serous or mucinous epithelia. Qualitative and quantitative analysis of the different imaging modalities used showed that multimodal nonlinear microscopy has the potential to differentiate between cancerous and healthy ovarian tissue.

  17. Conjugates of folic acids with BSA-coated quantum dots for cancer cell targeting and imaging by single-photon and two-photon excitation.

    Science.gov (United States)

    Meng, He; Chen, Ji-Yao; Mi, Lan; Wang, Pei-Nan; Ge, Mei-Ying; Yue, Yang; Dai, Ning

    2011-01-01

    Bovine serum albumin (BSA)-coated CdTe/ZnS quantum dots (BSA-QDs) were selected to conjugate with folic acid (FA), forming FA-BSA-QDs. This study aims to develop these small FA-BSA-QDs (less than 10 nm) for the diagnosis of cancers in which the FA receptor (FR) is overexpressed. The enhancement of cellular uptake in FR-positive human nasopharyngeal carcinoma cells (KB cells) for FA-BSA-QDs was found by means of confocal fluorescence microscopy under single-photon and two-photon excitation. The uptake enhancement for FA-BSA-QDs was further evaluated by flow-cytometric analysis in 10(4) KB cells, and was about 3 times higher than for BSA-QDs on average. The uptake enhancement was suppressed when KB cells had been pretreated with excess FA, reflecting that the enhancement was mediated by the association of FR at cell membranes with FA-BSA-QDs. When human embryonic kidney cells (293T) (FR-negative cells) and KB cells, respectively, were incubated with FA-BSA-QDs (1 μM) for 40 min, the FA-BSA-QD uptake by 293T cells was much weaker than that by KB cells, demonstrating that FA-BSA-QDs could undergo preferential binding on FR-positive cancer cells. These characteristics suggest that FA-BSA-QDs are potential candidates for cancer diagnosis.

  18. Sub-Doppler two-photon-excitation Rydberg spectroscopy of atomic xenon: mass-selective studies of isotopic and hyperfine structure

    Science.gov (United States)

    Kono, Mitsuhiko; He, Yabai; Baldwin, Kenneth G. H.; Orr, Brian J.

    2016-03-01

    Mass-selective sub-Doppler two-photon excitation (TPE) spectroscopy is employed to resolve isotopic contributions for transitions to high-energy Rydberg levels of xenon in an atomic beam, using narrowband pulses of coherent ultraviolet light at 205-213 nm generated by nonlinear-optical conversion processes. Previous research (Kono et al 2013 J. Phys. B: At. Mol. Opt. Phys. 46 35401), has determined isotope energy shifts and hyperfine structure for 33 high-energy Rydberg levels of gas-phase xenon and accessed Rydberg levels at TPE energies in the range of 94 100-97 300 cm-1 with unprecedented spectroscopic resolution. The new isotopic-mass-resolved results were obtained by adding a pulsed free-jet atomic-beam source and a mass-selective time-of-flight detector to the apparatus in order to discern individual xenon isotopes and extract previously unresolved spectroscopic information. Resulting isotope energy shifts and hyperfine-coupling parameters are examined with regard to trends in principal quantum number n and in atomic angular-momentum quantum numbers, together with empirical and theoretical precedents for such trends.

  19. Evaluation of Injured Axons Using Two-Photon Excited Fluorescence Microscopy after Spinal Cord Contusion Injury in YFP-H Line Mice.

    Science.gov (United States)

    Horiuchi, Hideki; Oshima, Yusuke; Ogata, Tadanori; Morino, Tadao; Matsuda, Seiji; Miura, Hiromasa; Imamura, Takeshi

    2015-07-13

    Elucidation of the process of degeneration of injured axons is important for the development of therapeutic modules for the treatment of spinal cord injuries. The aim of this study was to establish a method for time-lapse observation of injured axons in living animals after spinal cord contusion injury. YFP (yellow fluorescent protein)-H transgenic mice, which we used in this study, express fluorescence in their nerve fibers. Contusion damage to the spinal cord at the 11th vertebra was performed by IH (Infinite Horizon) impactor, which applied a pressure of 50 kdyn. The damaged spinal cords were re-exposed during the observation period under anesthesia, and then observed by two-photon excited fluorescence microscopy, which can observe deep regions of tissues including spinal cord axons. No significant morphological change of injured axons was observed immediately after injury. Three days after injury, the number of axons decreased, and residual axons were fragmented. Seven days after injury, only fragments were present in the damaged tissue. No hind-limb movement was observed during the observation period after injury. Despite the immediate paresis of hind-limbs following the contusion injury, the morphological degeneration of injured axons was delayed. This method may help clarification of pathophysiology of axon degeneration and development of therapeutic modules for the treatment of spinal cord injury.

  20. The effect of polyunsaturated fatty acids on the homeostasis of yolk lipoprotein in C. elegans examined by CARS and two-photon excitation fluorescence (TPE-F) microscopy

    Science.gov (United States)

    Chen, Wei-Wen; Yi, Yung-Hsiang; Chien, Cheng-Hao; Hsiung, Kuei-Ching; Lin, Yi-Chun; Ma, Tian-Hsiang; Lo, Szecheng J.; Chang, Ta-Chau

    2016-03-01

    Yolk lipoprotein constitutes the major source of energy and the materials for synthesizing signaling factors for the development of oocytes and embryos in C. elegans. Polyunsaturated fatty acids (PUFAs) packed in yolk lipoprotein have been recently recognized as critical molecules for fertilization and reproduction.1 However, the relation between PUFAs and the homeostasis of yolk lipoprotein is not clear. Here we use coherent anti-Stokes Raman scattering (CARS) microscopy and two-photon excitation fluorescence (TPE-F) microscopy to examine the transportation of yolk lipoprotein. We demonstrate that CARS microscopy is a more sensitive method than the traditional Nile Red staining method in probing the abnormal accumulation of yolk lipoprotein in the body cavity of C. elegans. It is found that the accumulation of yolk lipoprotein is a time-dependent process. In addition, a negative correlation (r = -0.955) between reproductive aging and abnormal accumulation of yolk lipoprotein is established. We further examine wild-type, fat-1, and fat-2 worms with or without the expression of GFP-tagged yolk lipoprotein (VIT-2-GFP). Our data reveal that PUFAs have a positive effect on the synthesis and endocytosis of yolk lipoprotein, confirming the model proposed by Edmonds et al.2

  1. Two-Photon Flow Cytometry

    Science.gov (United States)

    Zhog, Cheng Frank; Ye, Jing Yong; Norris, Theodore B.; Myc, Andrzej; Cao, Zhengyl; Bielinska, Anna; Thomas, Thommey; Baker, James R., Jr.

    2004-01-01

    Flow cytometry is a powerful technique for obtaining quantitative information from fluorescence in cells. Quantitation is achieved by assuring a high degree of uniformity in the optical excitation and detection, generally by using a highly controlled flow such as is obtained via hydrodynamic focusing. In this work, we demonstrate a two-beam, two- channel detection and two-photon excitation flow cytometry (T(sup 3)FC) system that enables multi-dye analysis to be performed very simply, with greatly relaxed requirements on the fluid flow. Two-photon excitation using a femtosecond near-infrared (NIR) laser has the advantages that it enables simultaneous excitation of multiple dyes and achieves very high signal-to-noise ratio through simplified filtering and fluorescence background reduction. By matching the excitation volume to the size of a cell, single-cell detection is ensured. Labeling of cells by targeted nanoparticles with multiple fluorophores enables normalization of the fluorescence signal and thus ratiometric measurements under nonuniform excitation. Quantitative size measurements can also be done even under conditions of nonuniform flow via a two-beam layout. This innovative detection scheme not only considerably simplifies the fluid flow system and the excitation and collection optics, it opens the way to quantitative cytometry in simple and compact microfluidics systems, or in vivo. Real-time detection of fluorescent microbeads in the vasculature of mouse ear demonstrates the ability to do flow cytometry in vivo. The conditions required to perform quantitative in vivo cytometry on labeled cells will be presented.

  2. Evaluation of dual-wavelength excitation autofluorescence imaging of colorectal tumours with a high-sensitivity CMOS imager: a cross-sectional study.

    Science.gov (United States)

    Kominami, Yoko; Yoshida, Shigeto; Tanaka, Shinji; Miyaki, Rie; Sanomura, Yoji; Seo, Min-Woong; Kagawa, Keiichiro; Kawahito, Shoji; Arimoto, Hidenobu; Yamada, Kenji; Chayama, Kazuaki

    2015-09-02

    It is important to devise efficient and easy methods of detecting colorectal tumours to reduce mortality from colorectal cancer. Dual-wavelength excitation autofluorescence intensity can be used to visualize colorectal tumours. Therefore, we evaluated dual-wavelength excitation autofluorescence images of colorectal tumours obtained with a newly developed, high-sensitivity complementary metal-oxide-semiconductor (CMOS) imager. A total 107 colorectal tumours (44 adenomas, 43 adenocarcinomas with intramucosal invasion, and 20 sessile serrated adenoma/polyps [SSA/Ps]) in 98 patients who underwent endoscopic tumour resection were included. The specimens were irradiated with excitation light at 365 nm and 405 nm, and autofluorescence images measured with a 475 ± 25-nm band pass filter were obtained using a new, high-sensitivity CMOS imager. Ratio images (F365ex/F405ex) were created to evaluate the lesion brightness compared with that of normal mucosa, and specimens were categorized into a no signal or high signal group. Adenomas and adenocarcinomas were depicted in 87 ratio images, with 86.2% (n = 75) in the High signal group. SSA/P was depicted in 20 ratio images, with 70.0% (n = 14) in the High signal group. Dual-wavelength excitation autofluorescence images of colorectal tumours can be acquired using our high-sensitivity CMOS imager, and are useful in detecting colorectal tumours.

  3. Label-free imaging immune cells and collagen in atherosclerosis with two-photon and second harmonic generation microscopy

    Directory of Open Access Journals (Sweden)

    Chunqiang Li

    2016-01-01

    Full Text Available Atherosclerosis has been recognized as a chronic inflammation disease, in which many types of cells participate in this process, including lymphocytes, macrophages, dendritic cells (DCs, mast cells, vascular smooth muscle cells (SMCs. Developments in imaging technology provide the capability to observe cellular and tissue components and their interactions. The knowledge of the functions of immune cells and their interactions with other cell and tissue components will facilitate our discovery of biomarkers in atherosclerosis and prediction of the risk factor of rupture-prone plaques. Nonlinear optical microscopy based on two-photon excited autofluorescence and second harmonic generation (SHG were developed to image mast cells, SMCs and collagen in plaque ex vivo using endogenous optical signals. Mast cells were imaged with two-photon tryptophan autofluorescence, SMCs were imaged with two-photon NADH autofluorescence, and collagen were imaged with SHG. This development paves the way for further study of mast cell degranulation, and the effects of mast cell derived mediators such as induced synthesis and activation of matrix metalloproteinases (MMPs which participate in the degradation of collagen.

  4. Phasor analysis of multiphoton spectral images distinguishes autofluorescence components of in vivo human skin.

    Science.gov (United States)

    Fereidouni, Farzad; Bader, Arjen N; Colonna, Anne; Gerritsen, Hans C

    2014-08-01

    Skin contains many autofluorescent components that can be studied using spectral imaging. We employed a spectral phasor method to analyse two photon excited autofluorescence and second harmonic generation images of in vivo human skin. This method allows segmentation of images based on spectral features. Various structures in the skin could be distinguished, including Stratum Corneum, epidermal cells and dermis. The spectral phasor analysis allowed investigation of their fluorescence composition and identification of signals from NADH, keratin, FAD, melanin, collagen and elastin. Interestingly, two populations of epidermal cells could be distinguished with different melanin content.

  5. Two-photon excitation laser scanning microscopy of porcine nasal septal cartilage following Nd:YAG laser-mediated stress relaxation

    Science.gov (United States)

    Kim, Charlton C.; Wallace, Vincent P.; Rasouli, Alexandre; Coleno, Mariah L.; Dao, Xavier; Tromberg, Bruce J.; Wong, Brian J.

    2000-05-01

    Laser irradiation of hyaline cartilage result in stable shape changes due to temperature dependent stress relaxation. In this study, we determined the structural changes in chondrocytes within porcine nasal septal cartilage tissue over a 4-day period using a two-photon laser scanning microscope (TPM) following Nd:YAG laser irradiation (lambda equals 1.32 micrometer) using parameters that result in mechanical stress relaxation (6.0 W, 5.4 mm spot diameter). TPM excitation (780 nm) result in induction of fluorescence from endogenous agents such as NADH, NADPH, and flavoproteins in the 400 - 500 nm spectral region. During laser irradiation diffuse reflectance (from a probe HeNe laser, (lambda) equals 632.8 nm), surface temperature, and stress relaxation were measured dynamically. Each specimen received one, two, or three sequential laser exposures (average irradiation times of 5, 6, and 8 seconds). The cartilage reached a peak surface temperature of about 70 degrees Celsius during irradiation. Cartilage denatured in 50% EtOH (20 minutes) was used as a positive control. TPM was performed using a mode-locked 780 nm Titanium:Sapphire (Ti:Al203) beam with a, 63X, 1.2 N.A. water immersion objective (working distance of 200 mm) to detect the fluorescence emission from the chondrocytes. Images of chondrocytes were obtained at depths up to 150 microns (lateral resolution equals 35 micrometer X 35 micrometer). Images were obtained immediately following laser exposure, and also after 4 days in culture. In both cases, the irradiated and non-irradiated specimens do not show any discernible difference in general shape or auto fluorescence. In contrast, positive controls (immersed in 50% ethanol), show markedly increased fluorescence relative to both the native and irradiated specimens, in the cytoplasmic region.

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

    Directory of Open Access Journals (Sweden)

    Philipp Steven

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

  7. Imaging marine virus CroV and its host Cafeteria roenbergensis with two-photon microscopy

    Science.gov (United States)

    Cao, Bin; Chakraborty, Sayan; Sun, Wenqing; Aghvami, Seyedmohammadali; Fischer, Matthias G.; Qian, Wei; Xiao, Chuan; Li, Chunqiang

    2014-02-01

    We use two-photon microscopy to monitor the infection process of marine zooplankton, Cafeteria roenbergensis (C.roenbergensis), by Cafeteria roenbergensis virus (CroV), a giant DNA virus named after its host. Here, we image C.roenbergensis in culture by two-photon excited NADH autofluorescence at video-rate (30 frame/s), and the movement of C.roenbergensis is recorded in live videos. Moreover, CroV is stained with DNA dye SYBR gold and recorded simultaneously with this two-photon microscope. We observed the initial infection moment with this method. The result demonstrates the potential use of two-photon microscopy to investigate the fast dynamic interaction between C.roenbergensis with virus CroV. After catching this initial moment, we will freeze the sample in liquid nitrogen for cryo-electron microscopy (EM) study to resolve the virus-host interaction at molecular level. The long-term goal is to study similar fast moving pathogen-host interaction process which could lead to important medical applications.

  8. Two-photon imaging and spectroscopy of fresh human colon biopsies

    Science.gov (United States)

    Cicchi, R.; Sturiale, A.; Nesi, G.; Tonelli, F.; Pavone, F. S.

    2012-03-01

    Two-photon fluorescence (TPEF) microscopy is a powerful tool to image human tissues up to 200 microns depth without any exogenously added probe. TPEF can take advantage of the autofluorescence of molecules intrinsically contained in a biological tissue, as such NADH, elastin, collagen, and flavins. Two-photon microscopy has been already successfully used to image several types of tissues, including skin, muscles, tendons, bladder. Nevertheless, its usefulness in imaging colon tissue has not been deeply investigated yet. In this work we have used combined two-photon excited fluorescence (TPEF), second harmonic generation microscopy (SHG), fluorescence lifetime imaging microscopy (FLIM), and multispectral two-photon emission detection (MTPE) to investigate different kinds of human ex-vivo fresh biopsies of colon. Morphological and spectroscopic analyses allowed to characterize both healthy mucosa, polyp, and colon samples in a good agreement with common routine histology. Even if further analysis, as well as a more significant statistics on a large number of samples would be helpful to discriminate between low, mild, and high grade cancer, our method is a promising tool to be used as diagnostic confirmation of histological results, as well as a diagnostic tool in a multiphoton endoscope or colonoscope to be used in in-vivo imaging applications.

  9. Fluorescence microscopy of single autofluorescent proteins for cellular biology

    CERN Document Server

    Cognet, Laurent; Choquet, Daniel; Lounis, Brahim

    2002-01-01

    In this paper we review the applicability of autofluorescent proteins for single-molecule imaging in biology. The photophysical characteristics of several mutants of the Green Fluorescent Protein (GFP) and those of DsRed are compared and critically discussed for their use in cellular biology. The alternative use of two-photon excitation at the single-molecule level or Fluorescence Correlation Spectroscopy is envisaged for the study of individual autofluorescent proteins. Single-molecule experiments performed in live cells using eGFP and preferably eYFP fusion proteins are reviewed. Finally, the first use at the single-molecule level of citrine, a more photostable variant of the eYFP is reported when fused to a receptor for neurotransmitter in live cells.

  10. Light-harvesting ytterbium(III)-porphyrinate-BODIPY conjugates: synthesis, excitation-energy transfer, and two-photon-induced near-infrared-emission studies.

    Science.gov (United States)

    Zhang, Tao; Zhu, Xunjin; Wong, Wai-Kwok; Tam, Hoi-Lam; Wong, Wai-Yeung

    2013-01-07

    Based on a donor-acceptor framework, several conjugates have been designed and prepared in which an electron-donor moiety, ytterbium(III) porphyrinate (YbPor), was linked through an ethynyl bridge to an electron-acceptor moiety, boron dipyrromethene (BODIPY). Photoluminescence studies demonstrated efficient energy transfer from the BODIPY moiety to the YbPor counterpart. When conjugated with the YbPor moiety, the BODIPY moiety served as an antenna to harvest the lower-energy visible light, subsequently transferring its energy to the YbPor counterpart, and, consequently, sensitizing the Yb(III) emission in the near-infrared (NIR) region with a quantum efficiency of up to 0.73% and a lifetime of around 40 μs. Moreover, these conjugates exhibited large two-photon-absorption cross-sections that ranged from 1048-2226 GM and strong two-photon-induced NIR emission.

  11. Two-Photon Irradiation of an Intracellular Singlet Oxygen Photosensitizer: Achieving Localized Sub-Cellular Excitation in Spatially-Resolved Experiments

    DEFF Research Database (Denmark)

    Pedersen, Brian Wett; Breitenbach, Thomas; Redmond, Robert W.;

    2010-01-01

    The response of a given cell to spatially-resolved sub-cellular irradiation of a singlet oxygen photosensitizer (protoporphyrin IX, PpIX) using a focused laser was assessed. In these experiments, incident light was scattered over a volume greater than that defi ned by the dimensions of the laser...... beam as a consequence of the inherent inhomogeneity of the cell. Upon irradiation at a wavelength readily absorbed by PpIX in a one-photon transition, this scattering of light eliminated any advantage accrued to the use of focused irradiation. However, upon irradiation at a longer wavelength where Pp......IX can only absorb light under non-linear two-photon conditions, meaningful intracellular resolution was achieved in the small spatial domain where the light intensity was high enough for absorption to occur....

  12. Excited-state dynamics and two-photon absorption cross sections of fluorescent diphenyl-tin(IV) derivatives with schiff bases: a comparative study of the effect of chelation from the ultrafast to the steady-state time scale.

    Science.gov (United States)

    Zugazagoitia, Jimena S; Maya, Mauricio; Damián-Zea, Carlos; Navarro, Pedro; Beltrán, Hiram I; Peon, Jorge

    2010-01-21

    Schiff bases bearing an intramolecular hydrogen bond are known to undergo excited-state intramolecular proton transfer and E-Z isomerization, which are related to their thermochromism and solvatochromism properties. In this study, we explored these ultrafast photoinduced processes for two doubly hydroxylated Schiff bases, salicylidene-2-aminophenol and 2-hydroxynaphthylmethylidene-2-aminophenol. From comparisons with our previously reported results for the parent monohidroxylated Schiff base salicylideneaniline, we were able to establish the lack of an effect of a second intramolecular hydrogen bond in the excited-state intramolecular proton-transfer process. Moreover, we synthesized and studied the photophysics of 14 diphenyl-tin(IV) derivatives with Schiff bases with the same framework as the former two. In these organometallic compounds, we observed an increase of more than 50 times in the excited-state decay times in comparison with those of the free ligands. This finding is attributed to the coordination with the metallic center, which restricts the fluctuations of the geometry of the organic Schiff base skeleton. The emission bands of these complexes can be easily tuned through substitutions at the Schiff base ligand and can be made to be centered well above 600 nm. The much enhanced emissive behavior of all diphenyl-tin(IV) derivatives allowed the study of several properties of their electronically excited states, including the effects of different substituents on their femtosecond and picosecond dynamics. Considering potential applications, we also performed transient absorption experiments to assess the wavelength interval for stimulated emission of this type of compound. Finally, we determined their two-photon absorption cross sections in the 760-820-nm range by measuring their two-photon induced fluorescence excitation spectra. Mainly, our results illustrate that the diphenyl-tin(IV) moiety, thanks to its size and its coordination mode with a single

  13. Two-photon imaging of stem cells

    Science.gov (United States)

    Uchugonova, A.; Gorjup, E.; Riemann, I.; Sauer, D.; König, K.

    2008-02-01

    A variety of human and animal stem cells (rat and human adult pancreatic stem cells, salivary gland stem cells, dental pulpa stem cells) have been investigated by femtosecond laser 5D two-photon microscopy. Autofluorescence and second harmonic generation have been imaged with submicron spatial resolution, 270 ps temporal resolution, and 10 nm spectral resolution. In particular, NADH and flavoprotein fluorescence was detected in stem cells. Major emission peaks at 460nm and 530nm with typical mean fluorescence lifetimes of 1.8 ns and 2.0 ns, respectively, were measured using time-correlated single photon counting and spectral imaging. Differentiated stem cells produced the extracellular matrix protein collagen which was detected by SHG signals at 435 nm.

  14. Two-photon microscopy for chemical neuroscience.

    Science.gov (United States)

    Ellis-Davies, Graham C R

    2011-04-20

    Microscopes using non-linear excitation of chromophores with pulsed near-IR light can generate highly localized foci of molecules in the electronic singlet state that are concentrated in volumes of less than one femtoliter. The three-dimensional confinement of excitation arises from the simultaneous absorption of two IR photons of approximately half the energy required for linear excitation. Two-photon microscopy is especially useful for two types of interrogation of neural processes. First, uncaging of signaling molecules such as glutamate, as stimulation is so refined it can be used to mimic normal unitary synaptic levels. In addition, uncaging allows complete control of the timing and position of stimulation, so the two-photon light beam provides the chemical neuroscientist with an "optical conductor's baton" which can command synaptic activity at will. A second powerful feature of two-photon microscopy is that when used for fluorescence imaging it enables the visualization of cellular structure and function in living animals at depths far beyond that possible with normal confocal microscopes. In this review I provide a survey of the many important applications of two-photon microscopy in these two fields of neuroscience, and suggest some areas for future technical development.

  15. Autofluorescence lifetime variation in the cuticle of the bedbug Cimex lectularius.

    Science.gov (United States)

    Reinhardt, Klaus; Breunig, Hans Georg; König, Karsten

    2017-01-01

    The decay time of the fluorescence of excited molecules, called fluorescence lifetime, can provide information about the cuticle composition additionally to widely used spectral characteristics. We compared autofluorescence lifetimes of different cuticle regions in the copulatory organ of females of the bedbug, Cimex lectularius. After two-photon excitation at 720 nm, regions recently characterised as being rich in resilin showed a longer bimodal distribution of the mean autofluorescence lifetime τm (tau-m) at 0.4 ns and 1.0-1.5 ns, while resilin-poor sites exhibited a unimodal pattern with a peak around 0.8 ns. The mean lifetime, and particularly its second component, can be useful to distinguish resilin-rich from resilin-poor parts of the cuticle. The few existing literature data suggest that chitin is unlikely responsible for the main autofluorescent component observed in the resilin-poor areas in our study and that melanin requires further scrutiny. Autofluorescence lifetime measurements can help to characterise properties of the arthropod cuticle, especially when coupled with multiphoton excitation to allow for deeper tissue penetration.

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

    Science.gov (United States)

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

    2011-07-01

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

  17. Magnetic two-photon scattering and two-photon emission - Cross sections and redistribution functions

    Science.gov (United States)

    Alexander, S. G.; Meszaros, P.

    1991-01-01

    The magnetic two-photon scattering cross section is discussed within the framework of QED, and the corresponding scattering redistribution function for this process and its inverse, as well as the scattering source function are calculated explicitly. In a similar way, the magnetic two-photon emission process which follows the radiative excitation of Landau levels above ground is calculated. The two-photon scattering and two-photon emission are of the same order as the single-photon magnetic scattering. All three of these processes, and in optically thick cases also their inverses, are included in radiative transport calculations modeling accreting pulsars and gamma-ray bursters. These processes play a prominent role in determining the relative strength of the first two cyclotron harmonics, and their effects extend also to the higher harmonics.

  18. Ultrafast dynamics of free carriers induced by two-photon excitation in bulk ZnSe crystal%双光子激发ZnSe自由载流子超快动力学研究∗

    Institute of Scientific and Technical Information of China (English)

    2015-01-01

    Semiconductor materials exhibiting large optical nonlinearities and ultrafast nonlinear response have received ex-tensive attention because of their potential applications in optical limiting, all-optical devices, optical telecommunication, and so on. As a direct-gap II-VI bulk semiconductor, ZnSe crystal has been exploited as the nonlinear optical devices in the regimes of nanoseconds and picoseconds during the past years. Owing to today’s fast advance of laser sources with ultrashort femtosecond pulse duration, it is possible to investigate the ultrafast optical nonlinearities in the bulk ZnSe crystal. In this paper, we experimentally investigate the ultrafast dynamics of free-carriers induced by two-photon excitation in the bulk ZnSe crystal. By performing open-aperture Z-scan experiments with 41 fs laser pulses at the wavelength of 532 nm under the condition of low excitation intensity, the two-photon absorption coefficient is measured. As the excitation intensity exceeds a critical value, the interplay between third- and fifth-order nonlinear absorption processes is observed. To evaluate the ultrafast dynamics of free carriers, we have carried out femtosecond time-resolved degen-erate pump-probe measurements with the same laser system used for Z-scan experiments in different levels of pump intensities. It is shown that the transient absorption signals peaked at the zero delay is a linearly increasing function of pump intensity, indicating that the observed instantaneous nonlinear absorption is dominated by the interband two-photon absorption process. At moderate irradiance, the transient absorption signals obviously indicate two components, arising from the two-photon absorption-induced free-carrier absorption, which is equivalent to the fifth-order nonlinear absorption process. Under the excitation of relatively high pump intensity, the magnitude of the reduction of free-carrier absorption signal becomes faster, suggesting that the ZnSe crystal exhibits a

  19. Generation of Terahertz Radiation in LED Heterostructures with Multiple InGaN/GaN Quantum Wells at Two-Photon Excitation by Femtosecond

    Science.gov (United States)

    Prudaev, I. A.; Sarkisov, S. Yu.; Tolbanov, O. P.; Kosobutsky, A. V.

    2015-06-01

    The results of experiments on the generation of terahertz radiation in the nitride LED structures at optical excitation by ultrashort laser pulses are presented. The dependences of the emission spectra on the structural parameters of samples and intensity of laser pulses are studied. An increase in the amplitude and the shift of the frequency spectra of terahertz pulses to higher frequencies with increasing number of quantum wells in the heterostructure is found.

  20. Two-photon absorption in arsenic sulfide glasses

    Science.gov (United States)

    Chunaev, D. S.; Snopatin, G. E.; Plotnichenko, V. G.; Karasik, A. Ya.

    2016-10-01

    The two-photon absorption coefficient of 1047-{\\text{nm}} light in {\\text{As}}35{\\text{S}}65 chalcogenide glass has been measured. CW probe radiation has been used to observe the linear absorption in glass induced by two-photon excitation. The induced absorption lifetime was found to be ∼ 2 {\\text{ms}}.

  1. Pulse-shaping based two-photon FRET stoichiometry.

    Science.gov (United States)

    Flynn, Daniel C; Bhagwat, Amar R; Brenner, Meredith H; Núñez, Marcos F; Mork, Briana E; Cai, Dawen; Swanson, Joel A; Ogilvie, Jennifer P

    2015-02-09

    Förster Resonance Energy Transfer (FRET) based measurements that calculate the stoichiometry of intermolecular interactions in living cells have recently been demonstrated, where the technique utilizes selective one-photon excitation of donor and acceptor fluorophores to isolate the pure FRET signal. Here, we present work towards extending this FRET stoichiometry method to employ two-photon excitation using a pulse-shaping methodology. In pulse-shaping, frequency-dependent phases are applied to a broadband femtosecond laser pulse to tailor the two-photon excitation conditions to preferentially excite donor and acceptor fluorophores. We have also generalized the existing stoichiometry theory to account for additional cross-talk terms that are non-vanishing under two-photon excitation conditions. Using the generalized theory we demonstrate two-photon FRET stoichiometry in live COS-7 cells expressing fluorescent proteins mAmetrine as the donor and tdTomato as the acceptor.

  2. Two-photon spectral fluorescence lifetime and second-harmonic generation imaging of the porcine cornea with a 12-femtosecond laser microscope

    Science.gov (United States)

    Batista, Ana; Breunig, Hans Georg; Uchugonova, Aisada; Morgado, António Miguel; König, Karsten

    2016-03-01

    Five dimensional microscopy with a 12-fs laser scanning microscope based on spectrally resolved two-photon autofluorescence lifetime and second-harmonic generation (SHG) imaging was used to characterize all layers of the porcine cornea. This setup allowed the simultaneous excitation of both metabolic cofactors, NAD(P)H and flavins, and their discrimination based on their spectral emission properties and fluorescence decay characteristics. Furthermore, the architecture of the stromal collagen fibrils was assessed by SHG imaging in both forward and backward directions. Information on the metabolic state and the tissue architecture of the porcine cornea were obtained with subcellular resolution, and high temporal and spectral resolutions.

  3. Two-photon physics

    Energy Technology Data Exchange (ETDEWEB)

    Mark Vanderhaeghen

    2005-10-22

    It is reviewed how Compton scattering sum rules relate low-energy nucleon structure quantities to the nucleon excitation spectrum. In particular, the GDH sum rule and recently proposed extensions of it will be discussed. These extensions are sometimes more calculationally robust, which may be an advantage when estimating the chiral extrapolations of lattice QCD results, such as for anomalous magnetic moments. Subsequently, new developments in our description of the nucleon excitation spectrum will be discussed, in particular a recently developed chiral effective field theory framework for the {Delta}(1232)-resonance region. Within this framework, we discuss results on N and {Delta} masses, the {gamma} N {Delta} transition and the {Delta} magnetic dipole moment.

  4. Two photon physics. Personal recollection

    CERN Document Server

    Ginzburg, Ilya F

    2015-01-01

    The term two--photon processes is used for the reactions in which some system of particles is produced in collision of two photons, either real or virtual. In the study of these processes our main goal was to suggest approach, allowing to extract from the data information on proper two--photon process separating it from mechanism which responsible for the production of photons. Here I present my view for history of two--photon physics. I don't try to give complete review, concentrating mainly on works of our team (which cover essential part of the topic) and some colleagues. My citation is strongly incomplete. I cite here only papers which were essential in our understanding of the problems. The choice of presented details is the result of my discussions with Gleb Kotkin and Valery Serbo. 1. Prehistory. 2. Two photon processes at e^+e^- colliders. 3. Photon colliders. 4. Notes on physical program.

  5. Two-Photon Collective Atomic Recoil Lasing

    Directory of Open Access Journals (Sweden)

    James A. McKelvie

    2015-11-01

    Full Text Available We present a theoretical study of the interaction between light and a cold gasof three-level, ladder configuration atoms close to two-photon resonance. In particular, weinvestigate the existence of collective atomic recoil lasing (CARL instabilities in differentregimes of internal atomic excitation and compare to previous studies of the CARL instabilityinvolving two-level atoms. In the case of two-level atoms, the CARL instability is quenchedat high pump rates with significant atomic excitation by saturation of the (one-photoncoherence, which produces the optical forces responsible for the instability and rapid heatingdue to high spontaneous emission rates. We show that in the two-photon CARL schemestudied here involving three-level atoms, CARL instabilities can survive at high pump rateswhen the atoms have significant excitation, due to the contributions to the optical forces frommultiple coherences and the reduction of spontaneous emission due to transitions betweenthe populated states being dipole forbidden. This two-photon CARL scheme may form thebasis of methods to increase the effective nonlinear optical response of cold atomic gases.

  6. Evaluation of cryo-preserved skin tissues using two-photon microscopy

    Science.gov (United States)

    Riemann, Iris; Beier, Axel; Schwarz, Martin; Dörr, Daniel; Stracke, Frank; Zimmermann, Heiko

    2010-02-01

    If no fresh skin samples can be obtained or used, it is important for research and industries to have models and stored tissue samples as close to the native state as possible at disposal. One way to preserve tissues for a longer timeframe is to use deep freezing cryo-techniques. Unfortunately much damage can be induced during the cooling and the thawing processes like disruption of cells and extra-cellular matrices due to the formation of ice crystals. This could lead to a disturbance of the united cell structure up to the point of a loss of cell viability. Two-photon microscopy is able to gather information about cells and tissue components via excitation of the autofluorescence deep inside the sample with a high resolution in both, frozen and thawed states. It is possible to monitor the samples before and after and, important, observe events during the freezing process like the formation of ice crystals. To determine the state of skin tissues after slow rate freezing and the quick process of vitrification, the samples were examined with two-photon microscopy. To establish an optimized freezing-protocol for skin tissues, morphological changes, changes in autofluorescence of endogenous fluorophores (NADH, keratin, flavins, elastin) or changes in second harmonic generation of collagen fibres could provide information about the quality of the used freezing parameters and protective additives and lead to an optimized freezing-protocol with a new set of parameters to obtain mostly intact tissue samples. Multiphoton microscopy has been established as a useful tool for optical in situ quality control of frozen tissues.

  7. Multiphoton autofluorescence lifetime imaging of induced pluripotent stem cells

    Science.gov (United States)

    Uchugonova, Aisada

    2017-06-01

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

  8. The development of efficient two-photon singlet oxygen sensitizers

    DEFF Research Database (Denmark)

    Nielsen, Christian Benedikt

    The development of efficient two-photon singlet oxygen sensitizers is addressed focusing on organic synthesis. Photophysical measurements were carried out on new lipophilic molecules, where two-photon absorption cross sections and singlet oxygen quantumyields were measured. Design principles...... for making efficient two-photon singlet oxygen sensitizers were then constructed from these results. Charge-transfer in the excited state of the prepared molecules was shown to play a pivotal role in the generationof singlet oxygen. This was established through studies of substituent effects on both...... the singlet oxygen yield and the two-photon absorption cross section, where it was revealed that a careful balancing of the amount of charge transfer present in theexcited state of the sensitizer is necessary to obtain both a high singlet oxygen quantum yield and a high two-photon cross section. An increasing...

  9. Higgs Decay to Two Photons

    OpenAIRE

    Marciano, William J.; Zhang, Cen; Willenbrock, Scott

    2011-01-01

    The amplitude for Higgs decay to two photons is calculated in renormalizable and unitary gauges using dimensional regularization at intermediate steps. The result is finite, gauge independent, and in agreement with previously published results. The large Higgs mass limit is examined using the Goldstone-boson equivalence theorem as a check on the use of dimensional regularization and to explain the absence of decoupling.

  10. Excited-state kinetics of the carotenoid S//1 state in LHC II and two-photon excitation spectra of lutein and beta-carotene in solution Efficient Car S//1 yields Chl electronic energy transfer via hot S//1 states?

    CERN Document Server

    Walla, P J; Linden, Patricia A; Ohta, Kaoru

    2002-01-01

    The excited-state dynamics of the carotenoids (Car) in light- harvesting complex II (LHC II) of Chlamydomonas reinhardtii were studied by transient absorption measurements. The decay of the Car S //1 population ranges from similar to 200 fs to over 7 ps, depending on the excitation and detection wavelengths. In contrast, a 200 fs Car S//1 yields Chlorophyll (Chl) energy transfer component was the dominant time constant for our earlier two-photon fluorescence up- conversion measurements (Walla, P.J. ; et al. J. Phys. Chem. B 2000, 104, 4799-4806). We also present the two-photon excitation (TPE) spectra of lutein and beta-carotene in solution and compare them with the TPE spectrum of LHC II. The TPE-spectrum of LHC II has an onset much further to the blue and a width that is narrower than expected from comparison to the S//1 fluorescence of lutein and beta-carotene in solution. Different environments may affect the shape of the S//1 spectrum significantly. To explain the blue shift of the TPE spectrum and the d...

  11. Two-photon flow cytometer with laser scanning Bessel beams

    Science.gov (United States)

    Wang, Yongdong; Ding, Yu; Ray, Supriyo; Paez, Aurelio; Xiao, Chuan; Li, Chunqiang

    2016-03-01

    Flow cytometry is an important technique in biomedical discovery for cell counting, cell sorting and biomarker detection. In vivo flow cytometers, based on one-photon or two-photon excited fluorescence, have been developed for more than a decade. One drawback of laser beam scanning two-photon flow cytometer is that the two-photon excitation volume is fairly small due to the short Rayleigh range of a focused Gaussian beam. Hence, the sampling volume is much smaller than one-photon flow cytometry, which makes it challenging to count or detect rare circulating cells in vivo. Bessel beams have narrow intensity profiles with an effective spot size (FWHM) as small as several wavelengths, making them comparable to Gaussian beams. More significantly, the theoretical depth of field (propagation distance without diffraction) can be infinite, making it an ideal solution as a light source for scanning beam flow cytometry. The trade-off of using Bessel beams rather than a Gaussian beam is the fact that Bessel beams have small concentric side rings that contribute to background noise. Two-photon excitation can reduce this noise, as the excitation efficiency is proportional to intensity squared. Therefore, we developed a two-photon flow cytometer using scanned Bessel beams to form a light sheet that intersects the micro fluidic channel.

  12. Two-photon pumped lead halide perovskite nanowire lasers

    CERN Document Server

    Gu, Zhiyuan; Sun, Wenzhao; Li, Jinakai; Liu, Shuai; Song, Qinghai; Xiao, Shumin

    2015-01-01

    Solution-processed lead halide perovskites have shown very bright future in both solar cells and microlasers. Very recently, the nonlinearity of perovskites started to attract considerable research attention. Second harmonic generation and two-photon absorption have been successfully demonstrated. However, the nonlinearity based perovskite devices such as micro- & nano- lasers are still absent. Here we demonstrate the two-photon pumped nanolasers from perovskite nanowires. The CH3NH3PbBr3 perovskite nanowires were synthesized with one-step solution self-assembly method and dispersed on glass substrate. Under the optical excitation at 800 nm, two-photon pumped lasing actions with periodic peaks have been successfully observed at around 546 nm. The obtained quality (Q) factors of two-photon pumped nanolasers are around 960, and the corresponding thresholds are about 674?J=cm2. Both the Q factors and thresholds are comparable to conventional whispering gallery modes in two-dimensional polygon microplates. Ou...

  13. Direct Writing of Photonic Structures by Two-Photon Polymerization

    Directory of Open Access Journals (Sweden)

    Li Yan

    2013-11-01

    Full Text Available Single-mode dielectric-loaded surface plasmon-polariton nanowaveguides with strong mode confinement at excitation wavelength of 830 nm and high-Q polymer whispering gallery mode microcavities with surface roughness less than 12 nm have been directly written by two-photon polymerization, which pave the way to fabricate 3D plasmonic photonic structures by direct laser writing.

  14. A two-photon probe for Al(3+) in aqueous solution and its application in bioimaging.

    Science.gov (United States)

    Wang, Haihong; Wang, Bei; Shi, Zhaohua; Tang, Xiaoliang; Dou, Wei; Han, Qingxin; Zhang, Yange; Liu, Weisheng

    2015-03-15

    A salicylimine probe L with a simple structure has been researched more in-depth on fluorescence sensor properties based on two-photon (TP) absorption. L displays excellent selective turn-on fluorescence response for Al(3+) in hexamethylenetetramine-buffered (HMTA) aqueous solution (0.3M, pH=5.8) under one-photon (OP) excitation. With the help of OP fluorescence, TP fluorescence titration, UV-spectra titration and Job's plot, the stoichiometric ratio of L with Al(3+) was determined to be 1:1. The coordination sites and the coordination mechanism of L with Al(3+) were analyzed in detail through (1)H NMR data. Not only with a detection limit of 5.2×10(-9)M in vitro, but also the probe has been successfully used in the live cells and tissues for the imaging of Al(3+) with TP fluorescence microscopy due to the enlarged TP cross section, providing a novel testing method for measuring Al(3+) in solution or cell tissue with low autofluorescence and cytotoxicity.

  15. High contrast two-photon imaging of fingermarks

    Science.gov (United States)

    Stoltzfus, Caleb R.; Rebane, Aleksander

    2016-04-01

    Optically-acquired fingermarks are widely used as evidence across law enforcement agencies as well as in the courts of law. A common technique for visualizing latent fingermarks on nonporous surfaces consists of cyanoacrylate fuming of the fingerprint material, followed by impregnation with a fluorescent dye, which under ultra violet (UV) illumination makes the fingermarks visible and thus accessible for digital recording. However, there exist critical circumstances, when the image quality is compromised due to high background scattering, high auto-fluorescence of the substrate material, or other detrimental photo-physical and photo-chemical effects such as light-induced damage to the sample. Here we present a novel near-infrared (NIR), two-photon induced fluorescence imaging modality, which significantly enhances the quality of the fingermark images, especially when obtained from highly reflective and/or scattering surfaces, while at the same time reducing photo-damage to sensitive forensic samples.

  16. Three-dimensional microfabrication using two-photon polymerization

    Science.gov (United States)

    Cumpston, Brian H.; Ehrlich, Jeffrey E.; Kuebler, Stephen M.; Lipson, Matthew; Marder, Seth R.; McCord-Maughon, D.; Perry, Joseph W.; Roeckel, Harold; Rumi, Maria Cristina

    1998-09-01

    Photopolymerization initiated by the simultaneous absorption of two photons is unique in its ability to produce complex three-dimensional (3D) structures from a single, thick photopolymer film. Strong 3D confinement of the polymerization process is not possible in other polymer microfabrication techniques such as LIGA, rapid prototyping, and conventional photoresist technology. Two-photon polymerization also permits the fabrication of 3D structures and the definition of lithographic features on non-planar surfaces. We have developed a wide array of chromophores which hold great promise for 3D microfabrication, as well as other applications, such as two-photon fluorescence imaging and 3D optical data storage. These materials are based on a donor- (pi) -donor, donor-acceptor-donor, or acceptor-donor-acceptor structural motif. The magnitude of the two-photon absorption cross-section, (delta) , and the position of the two-photon absorption maximum, (lambda) (2)max, can be controlled by varying the length of the conjugated bridge and by varying the strength of the donor/acceptor groups. In this way, chromophores have been developed which exhibit strong two- photon absorption in the range of 500 - 975 nm, in some cases as high as 4400 X 10-50 cm4 s/photon-molecule. In the case of donor-(pi) -donor structures, quantum-chemical calculations show that the large absorption cross-sections arise from the symmetric re-distribution of charge from the donor end-groups to the conjugated bridge, resulting in an electronic excited-state which is more delocalized than the ground state. For many of these molecules, two-photon excitation populates a state which is sufficiently reducing that a charge transfer reaction can occur with acrylate monomers. The efficiency of these processes can be described using Marcus theory. Under suitable conditions, such reactions can induce radical polymerization of acrylate resins. Polymerization rates have been measured, and we show that these two-photon

  17. Enhanced two-photon absorption using true thermal light

    CERN Document Server

    Jechow, Andreas; Kurzke, Henning; Heuer, Axel; Menzel, Ralf

    2013-01-01

    Two-photon excited fluorescence (TPEF) is a standard technique in modern microscopy but still affected by photo-damage of the probe. It was proposed that TPEF can be enhanced by using entangled photons, but has proven to be challenging. Recently it was shown that some features of entangled photons can be mimicked with thermal light, which finds application in ghost imaging, sub-wavelength lithography and metrology. Here, we utilize true thermal light from a super-luminescence diode to demonstrate enhanced TPEF compared to coherent light using two common fluorophores and luminescent quantum dots. We find that the two-photon absorption rate is directly proportional to the measured degree of second-order coherence, as predicted by theory. Our results show that photon bunching can be exploited in two-photon microscopy with the photon statistic providing a new degree of freedom.

  18. Two-Photon Absorption of Metal-Assisted Chromophores.

    Science.gov (United States)

    Li, Xin; Rinkevicius, Zilvinas; Ågren, Hans

    2014-12-09

    Aiming to understand the effect of a metal surface on nonlinear optical properties and the combined effects of surface and solvent environments on such properties, we present a multiscale response theory study, integrated with dynamics of the two-photon absorption of 4-nitro-4'-amino-trans-stilbene physisorbed on noble metal surfaces, considering two such surfaces, Ag(111) and Au(111), and two solvents, cyclohexane and water, as cases for demonstration. A few conclusions of general character could be drawn: While the geometrical change of the chromophore induced by the environment was found to notably alter (diminish) the two-photon absorption cross section in the polar medium, the effects of the metal surface and solvent on the electronic structure of the chromophore surpasses the geometrical effects and leads to a considerably enhanced two-photon absorption cross section in the polar solvent. This enhancement of two-photon absorption arises essentially from the metal charge image induced enlargement of the difference between the dipole moment of the excited state and the ground state. The orientation-dependence of the two-photon absorption is found to connect with the lateral rotation of the chromophore, where the two-photon absorption reaches its maximum when the polarization of the incident light coincides with the long-axis of the chromophore. Our results demonstrate a distinct enhancement of the two-photon absorption by a metal surface and a polar medium and envisage the employment of metal-chromophore composite materials for future development of nonlinear optical materials with desirable properties.

  19. A Two- Photon Femtosecond Laser System for Three-Dimensional Microfabrication and Data Storage

    Institute of Scientific and Technical Information of China (English)

    蒋中伟; 周拥军; 袁大军; 黄文浩; 夏安东

    2003-01-01

    Utilizing the well-focused femtosecond laser with extreme high pulse intensity, we built a two-photon microfabrication and data storage system, which was introduced through several functional parts. Based on this homemade system, several three-dimensional microstructures were fabricated by two-photon polymerization, and three-dimensional data storage of six-layers was achieved by two-photon excitation with a photochromic material.

  20. Two-photon approximation in the theory of the electron recombination in hydrogen

    OpenAIRE

    Solovyev, D.; Labzowsky, L.

    2010-01-01

    A rigorous QED theory of the multiphoton decay of excited states in hydrogen atom is presented. The "two-photon" approximation is formulated which is limited by the one-photon and two-photon transitions including cascades transitions with two-photon links. This may be helpful for the strict description of the recombination process in hydrogen atom and, in principle, for the history of the hydrogen recombination in the early Universe.

  1. Confocal and Two-Photon Microscopy: Foundations, Applications and Advances

    Science.gov (United States)

    Diaspro, Alberto

    2001-11-01

    Confocal and Two-Photon Microscopy Foundations, Applications, and Advances Edited by Alberto Diaspro Confocal and two-photon fluorescence microscopy has provided researchers with unique possibilities of three-dimensional imaging of biological cells and tissues and of other structures such as semiconductor integrated circuits. Confocal and Two-Photon Microscopy: Foundations, Applications, and Advances provides clear, comprehensive coverage of basic foundations, modern applications, and groundbreaking new research developments made in this important area of microscopy. Opening with a foreword by G. J. Brakenhoff, this reference gathers the work of an international group of renowned experts in chapters that are logically divided into balanced sections covering theory, techniques, applications, and advances, featuring: In-depth discussion of applications for biology, medicine, physics, engineering, and chemistry, including industrial applications Guidance on new and emerging imaging technology, developmental trends, and fluorescent molecules Uniform organization and review-style presentation of chapters, with an introduction, historical overview, methodology, practical tips, applications, future directions, chapter summary, and bibliographical references Companion FTP site with full-color photographs The significant experience of pioneers, leaders, and emerging scientists in the field of confocal and two-photon excitation microscopy Confocal and Two-Photon Microscopy: Foundations, Applications, and Advances is invaluable to researchers in the biological sciences, tissue and cellular engineering, biophysics, bioengineering, physics of matter, and medicine, who use these techniques or are involved in developing new commercial instruments.

  2. Holographic Two-Photon Induced Photopolymerization

    Data.gov (United States)

    Federal Laboratory Consortium — Holographic two-photon-induced photopolymerization (HTPIP) offers distinct advantages over conventional one-photon-induced photopolymerization and current techniques...

  3. Two-photon STED spectral determination for a new V-shaped organic fluorescent probe with efficient two-photon absorption.

    Science.gov (United States)

    Belfield, Kevin D; Bondar, Mykhailo V; Morales, Alma R; Padilha, Lazaro A; Przhonska, Olga V; Wang, Xuhua

    2011-10-24

    Two-photon stimulated emission depletion (STED) cross sections were determined over a broad spectral range for a novel two-photon absorbing organic molecule, representing the first such report. The synthesis, comprehensive linear photophysical, two-photon absorption (2PA), and stimulated emission properties of a new fluorene-based compound, (E)-2-{3-[2-(7-(diphenylamino)-9,9-diethyl-9H-fluoren-2-yl)vinyl]-5-methyl-4-oxocyclohexa-2,5-dienylidene} malononitrile (1), are presented. Linear spectral parameters, including excitation anisotropy and fluorescence lifetimes, were obtained over a broad range of organic solvents at room temperature. The degenerate two-photon absorption (2PA) spectrum of 1 was determined with a combination of the direct open-aperture Z-scan and relative two-photon-induced fluorescence methods using 1 kHz femtosecond excitation. The maximum value of the 2PA cross section ~1700 GM was observed in the main, long wavelength, one-photon absorption band. One- and two-photon stimulated emission spectra of 1 were obtained over a broad spectral range using a femtosecond pump-probe technique, resulting in relatively high two-photon stimulated emission depletion cross sections (~1200 GM). A potential application of 1 in bioimaging was demonstrated through one- and two-photon fluorescence microscopy images of HCT 116 cells incubated with micelle-encapsulated dye.

  4. Synthesis of two carbazole-based dyes and application of two-photon initiating polymerization

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    Two carbazole-based polymerization initiators possessing blue fluorescence emission have been synthesized via Wittig reaction in the solid phase at room temperature.Two-photon excited fluorescence(TPEF) spectra for them were investigated under 800 nm fs laser pulse and two-photon absorption cross sections were determined by the Z-scan technique.Then two-photon initiating polymerization(TPIP) microfabrication experiments were successfully carried out.Three-dimensional lattice and artificial defects were gained,indicating that they were viable candidates for the two-photon polymerization initiator in practical application of microfabrication.

  5. Synthesis of two carbazole-based dyes and application of two-photon initiating polymerization

    Institute of Scientific and Technical Information of China (English)

    HU RenTao; LU LiangFei; RUAN BanFeng; WANG Peng; ZHANG MingLiang; ZHOU HongPing; LI ShengLi; WU JieYing; TIAN YuPeng

    2009-01-01

    Two carbazole-based polymerization initiators possessing blue fluorescence emission have been synthesized via Wittig reaction in the solid phase at room temperature.Two-photon excited fluorescence (TPEF) spectra for them were investigated under 800 nm fs laser pulse and two-photon absorption cross sections were determined by the Z-scan technique.Then two-photon initiating polymerization (TPIP) microfabrication experiments were successfully carried out.Three-dimensional lattice and artificial defects were gained,indicating that they were viable candidates for the two-photon polymerization initiator in practical application of microfabrication.

  6. A new approach to dual-color two-photon microscopy with fluorescent proteins

    Directory of Open Access Journals (Sweden)

    Rebane Aleks

    2010-02-01

    Full Text Available Abstract Background Two-photon dual-color imaging of tissues and cells labeled with fluorescent proteins (FPs is challenging because most two-photon microscopes only provide one laser excitation wavelength at a time. At present, methods for two-photon dual-color imaging are limited due to the requirement of large differences in Stokes shifts between the FPs used and their low two-photon absorption (2PA efficiency. Results Here we present a new method of dual-color two-photon microscopy that uses the simultaneous excitation of the lowest-energy electronic transition of a blue fluorescent protein and a higher-energy electronic transition of a red fluorescent protein. Conclusion Our method does not require large differences in Stokes shifts and can be extended to a variety of FP pairs with larger 2PA efficiency and more optimal imaging properties.

  7. Imaging theory and resolution improvement of two-photon confocal microscopy

    Institute of Scientific and Technical Information of China (English)

    唐志列; 杨初平; 裴红津; 梁瑞生; 刘颂豪

    2002-01-01

    The nonlinear effect of two-photon excitation on the imaging property of two-photonconfocal microscopy has been analyzed by the two-photon fluorescence intensity transfer functionderived in this paper. The two-photon fluorescence intensity transfer function in a confocal micros-copy is given. Furthermore the three-dimensional point spread function (3D-PSF) and thethree-dimensional optical transfer function (3D-OTF) of two-photon confocal microscopy are de-rived based on the nonlinear effect of two-photon excitation. The imaging property of two-photonconfocal microscopy is discussed in detail based on 3D-OTF. Finally the spatial resolution limit oftwo-photon confocal microscopy is discussed according to the uncertainty principle.

  8. Two-Photon-Pumped Perovskite Semiconductor Nanocrystal Lasers.

    Science.gov (United States)

    Xu, Yanqing; Chen, Qi; Zhang, Chunfeng; Wang, Rui; Wu, Hua; Zhang, Xiaoyu; Xing, Guichuan; Yu, William W; Wang, Xiaoyong; Zhang, Yu; Xiao, Min

    2016-03-23

    Two-photon-pumped lasers have been regarded as a promising strategy to achieve frequency up-conversion for situations where the condition of phase matching required by conventional approaches cannot be fulfilled. However, their practical applications have been hindered by the lack of materials holding both efficient two-photon absorption and ease of achieving population inversion. Here, we show that this challenge can be tackled by employing colloidal nanocrystals of perovskite semiconductors. We observe highly efficient two-photon absorption (with a cross section of 2.7 × 10(6) GM) in toluene solutions of CsPbBr3 nanocrystals that can excite large optical gain (>500 cm(-1)) in thin films. We have succeeded in demonstrating stable two-photon-pumped lasing at a remarkable low threshold by coupling CsPbBr3 nanocrystals with microtubule resonators. Our findings suggest perovskite nanocrystals can be used as excellent gain medium for high-performance frequency-up-conversion lasers toward practical applications.

  9. Two-photon absorbing porphyrins for oxygen microscopy (Conference Presentation)

    Science.gov (United States)

    Esipova, Tatiana V.; Vinogradov, Sergei A.

    2016-03-01

    The ability to quantify oxygen in vivo in 3D with high spatial and temporal resolution is invaluable for many areas of the biomedical science, including ophthalmology, neuroscience, cancer and stem biology. An optical method based on oxygen-dependent quenching of phosphorescence is being developed, that allows quantitative minimally invasive real-time imaging of partial pressure of oxygen (pO2) in tissue. In the past, dendritically protected phosphorescent oxygen probes with controllable quenching parameters and defined bio-distributions have been developed. More recently our probe strategy has extended to encompass two-photon excitable oxygen probes, which brought about first demonstrations of two-photon phosphorescence lifetime microscopy (2PLM) of oxygen in vivo, providing new valuable information for neuroscience and stem cell biology. However, current two-photon oxygen probes suffer from a number of limitations, such as low brightness and high cost of synthesis, which dramatically reduce imaging performance and limit usability of the method. Here we present an approach to new bright phosphorescent chromophores with internally enhanced two-photon absorption cross-sections, which pave a way to novel proves for 2PLM. In addition to substantial increase in performance, the new probes can be synthesized by much more efficient methods, thereby greatly reducing the cost of the synthesis and making the technique accessible to a broader range of researchers across different fields.

  10. Fano interference in two-photon transport

    Science.gov (United States)

    Xu, Shanshan; Fan, Shanhui

    2016-10-01

    We present a general input-output formalism for the few-photon transport in multiple waveguide channels coupled to a local cavity. Using this formalism, we study the effect of Fano interference in two-photon quantum transport. We show that the physics of Fano interference can manifest as an asymmetric spectral line shape in the frequency dependence of the two-photon correlation function. The two-photon fluorescence spectrum, on the other hand, does not exhibit the physics of Fano interference.

  11. Effect of metalloporphyrins on red autofluorescence from oral bacteria.

    Science.gov (United States)

    Volgenant, Catherine M C; van der Veen, Monique H; de Soet, Johannes J; ten Cate, Jacob M

    2013-06-01

    The aim of this study was to assess the red autofluorescence from bacterial species related to dental caries and periodontitis in the presence of different nutrients in the growth medium. Bacteria were grown anaerobically on tryptic soy agar (TSA) supplemented with nutrients, including magnesium-porphyrins from spinach and iron-porphyrins from heme. The autofluorescence was then assessed at 405 nm excitation. On the TSA without additives, no autofluorescence was observed from any of the species tested. On the TSA containing sheep blood, red autofluorescence was observed only from Parvimonas micra. When the TSA was supplemented with blood, hemin, and vitamin K, red autofluorescence was observed from Actinomyces naeslundii, Bifidobacterium dentium, and Streptococcus mutans. Finally, on the TSA supplemented with spinach extract, red autofluorescence was observed from Aggregatibacter actinomycetemcomitans, A. naeslundii, Enterococcus faecalis, Fusobacterium nucleatum, Lactobacillus salivarius, S. mutans, and Veillonella parvula. We conclude that the bacteria related to dental caries and periodontal disease exhibit red autofluorescence. The autofluorescence characteristics of the tested strains depended on the nutrients present, such as metalloporphyrins, suggesting that the metabolic products of the oral biofilm could be responsible for red autofluorescence. © 2013 Eur J Oral Sci.

  12. Autofluorescence of viable cultured mammalian cells.

    Science.gov (United States)

    Aubin, J E

    1979-01-01

    The autofluorescence other than intrinsic protein emission of viable cultured mammalian cells has been investigated. The fluorescence was found to originate in discrete cytoplasmic vesicle-like regions and to be absent from the nucleus. Excitation and emission spectra of viable cells revealed at least two distinct fluorescent species. Comparison of cell spectra with spectra of known cellular metabolites suggested that most, if not all, of the fluorescence arises from intracellular nicotinamide adenine dinucleotide (NADH) and riboflavin and flavin coenzymes. Various changes in culture conditions did not affect the observed autofluorescence intensity. A multiparameter flow system (MACCS) was used to compare the fluorescence intensities of numerous cultured mammalian cells.

  13. Adiabatic following in two-photon transition

    Energy Technology Data Exchange (ETDEWEB)

    Nayfeh, M.H.; Nayfeh, A.H.

    1977-01-01

    There has been much interest recently in coherent multiphoton transitions in many-level systems. The present work considers the effect of relaxation in the response of a three-level system to a smoothly varying, near-resonant, two-photon field. The relaxation-dependent contributions to the nonlinear refractive index are calculated. It is shown that the coherent interaction of two smoothly varying, near-resonant, two-photon pulses with a three-level system can be described by ''two-photon damped Bloch equations'' which are analogous to those for a one-photon transition in a two-level system except for the presence of a two-photon coupling and a frequency shift. 1 figure. (RWR)

  14. Two-Photon Physics in Hadronic Processes

    Energy Technology Data Exchange (ETDEWEB)

    Carl Carlson; Marc Vanderhaeghen

    2007-11-01

    Two-photon exchange contributions to elastic electron-scattering are reviewed. The apparent discrepancy in the extraction of elastic nucleon form factors between unpolarized Rosenbluth and polarization transfer experiments is discussed, as well as the understanding of this puzzle in terms of two-photon exchange corrections. Calculations of such corrections both within partonic and hadronic frameworks are reviewed. In view of recent spin-dependent electron scattering data, the relation of the two-photon exchange process to the hyperfine splitting in hydrogen is critically examined. The imaginary part of the two-photon exchange amplitude as can be accessed from the beam normal spin asymmetry in elastic electron-nucleon scattering is reviewed. Further extensions and open issues in this field are outlined.

  15. Sideband-Induced Two-Photon Transparency

    Institute of Scientific and Technical Information of China (English)

    CHENG Guang-Ling; HU Xiang-Ming

    2006-01-01

    @@ We show that it is possible to use a single sideband to induce two-photon transparency in a three-level cascade medium. The medium simultaneously absorbs two photons as a one-step process when the middle level is far off one-photon resonance. A resonant sideband coupling on the upper transition and the two-photon one-step process drive the medium into a trapped state, and the dominant component is the ground state. Thus almost all population is trapped in the ground state and the two-photon absorption is dramatically suppressed. We present a numerical calculation for arbitrary values of the atomic and field parameters and also provide an analytic description for the required conditions.

  16. Two-photon induced photoluminescence and singlet oxygen generation from aggregated gold nanoparticles.

    Science.gov (United States)

    Jiang, Cuifeng; Zhao, Tingting; Yuan, Peiyan; Gao, Nengyue; Pan, Yanlin; Guan, Zhenping; Zhou, Na; Xu, Qing-Hua

    2013-06-12

    Metal nanoparticles have potential applications as bioimaging and photosensitizing agents. Aggregation effects are generally believed to be adverse to their biomedical applications. Here we have studied the aggregation effects on two-photon induced photoluminescence and singlet oxygen generation of Au nanospheres and Au nanorods of two different aspect ratios. Aggregated Au nanospheres and short Au nanorods were found to display enhanced two-photon induced photoluminescence and singlet oxygen generation capabilities compared to the unaggregated ones. The two-photon photoluminescence of Au nanospheres and short Au nanorods were enhanced by up to 15.0- and 2.0-fold upon aggregation, and the corresponding two-photon induced singlet oxygen generation capabilities were enhanced by 8.3 and 1.8-fold, respectively. The two-photon induced photoluminescence and singlet oxygen generation of the aggregated long Au nanorods were found to be lower than the unaggregated ones. These results support that the change in their two-photon induced photoluminescence and singlet oxygen generation originate from aggregation modulated two-photon excitation efficiency. This finding is expected to foster more biomedical applications of metal nanoparticles as Au nanoparticles normally exist in an aggregated form in the biological environments. Considering their excellent biocompatibility, high inertness, ready conjugation, and easy preparation, Au nanoparticles are expected to find more applications in two-photon imaging and two-photon photodynamic therapy.

  17. Correlations of two photons at hadron colliders

    OpenAIRE

    Kozlov, G. A.

    2011-01-01

    We study the Bose-Einstein correlations of two photons and their coherent properties that can provide the information about the space-time structure of the emitting source through the Higgs-boson decays into two photons. We argue that such an investigation could probe the Higgs-boson mass. The model is rather sensitive to the temperature of the environment and to the external distortion effect in medium.

  18. Platinum Acetylide Two-Photon Chromophores (Preprint)

    Science.gov (United States)

    2007-04-01

    the higher energy range that lead to its photodegradation . Secondly, because there is a quadratic dependence of two-photon absorption (2PA) on the...to either an electron donating amino- fluorenyl or electron withdrawing benzothiazolyl-fluorene that are themselves known as two-photon absorbing dyes ...groups in place of phenyl groups have shown a doubling of the intrinsic cr2value at 740 nm.40,41In this paper we describe novel platinum dyes that

  19. Two-photon imaging through a multimode fiber

    CERN Document Server

    Morales-Delgado, Edgar E; Moser, Christophe

    2015-01-01

    In this work we demonstrate 3D imaging using two-photon excitation through a 20 cm long multimode optical fiber (MMF) of 350 micrometers diameter. The imaging principle is similar to single photon fluorescence through a MMF, except that a focused femtosecond pulse is delivered and scanned over the sample. In our approach, focusing and scanning through the fiber is accomplished by digital phase conjugation using mode selection by time gating with an ultra-fast reference pulse. The excited two-photon emission is collected through the same fiber. We demonstrate depth sectioning by scanning the focused pulse in a 3D volume over a sample consisting of fluorescent beads suspended in a polymer. The achieved resolution is 1 micrometer laterally and 15 micrometers axially. Scanning is performed over an 80x80 micrometers field of view. To our knowledge, this is the first demonstration of high-resolution three-dimensional imaging using two-photon fluorescence through a multimode fiber.

  20. New developments in two-photon analysis of human skin in vivo

    Science.gov (United States)

    Riemann, I.; Schwarz, M.; Stracke, F.; Ehlers, A.; Dimitrow, E.; Kaatz, M.; König, K.; Le Harzic, R.

    2009-02-01

    Two-photon imaging of human skin using ultra short laser pulses can be used to obtain information about the state of cells and tissues by means of their natural autofluorescence. Using this method, it is possible to determine whether the normal cell pattern is disturbed or the autofluorescence is influenced by internal or external stimuli. Two-photon fluorescence lifetime imaging (FLIM) can further enhance this providing information about physiological processes, fluorophores (like NAD(P)H, collagen, keratin, elastin, flavins, melanin,...) and external applied probes inside cells and tissue parts. For example the part of the cells metabolism and energy level can be determined by analyzing the NADH regarding its free / bound state and its oxidized / reduced state. The combination of two-photon imaging with FLIM may lead to a better understanding and diagnosis of skin reactions and disorders. We also present some results of in vivo simultaneous collagen and elastin measurements in skin dermis. Changes of dermal collagen and elastin content are characteristic for skin aging as well as for pathological skin conditions.

  1. Modulation of attosecond beating in resonant two-photon ionization

    CERN Document Server

    Galán, Álvaro J; Martín, Fernando

    2014-01-01

    We present a theoretical study of the photoelectron attosecond beating at the basis of RABBIT (Reconstruction of Attosecond Beating By Interference of Two-photon transitions) in the presence of autoionizing states. We show that, as a harmonic traverses a resonance, its sidebands exhibit a peaked phase shift as well as a modulation of the beating frequency itself. Furthermore, the beating between two resonant paths persists even when the pump and the probe pulses do not overlap, thus providing a sensitive non-holographic interferometric means to reconstruct coherent metastable wave packets. We characterize these phenomena quantitatively with a general finite-pulse analytical model that accounts for the effect of both intermediate and final resonances on two-photon processes, at a negligible computational cost. The model predictions are in excellent agreement with those of accurate ab initio calculations for the helium atom in the region of the N=2 doubly excited states.

  2. Simultaneous two-photon activation of type-I photodynamic therapy agents.

    Science.gov (United States)

    Fisher, W G; Partridge, W P; Dees, C; Wachter, E A

    1997-08-01

    The excitation and emission properties of several psoralen derivatives are compared using conventional single-photon excitation and simultaneous two-photon excitation (TPE). Two-photon excitation is effected using the output of a mode-locked titanium: sapphire laser, the near infrared output of which is used to promote nonresonant TPE directly. Specifically, the excitation spectra and excited-state properties of 8-methoxypsoralen and 4'-aminomethyl-4,5,8-trimethylpsoralen are shown to be equivalent using both modes of excitation. Further, in vitro feasibility of two-photon photodynamic therapy (PDT) is demonstrated using Salmonella typhimurium. Two-photon excitation may be beneficial in the practice of PDT because it would allow replacement of visible or UV excitation light with highly penetrating, nondamaging near infrared light and could provide a means for improving localization of therapy. Comparison of possible laser excitation sources for PDT reveals the titanium: sapphire laser to be exceptionally well suited for nonlinear excitation of PDT agents in biological systems due to its extremely short pulse width and high repetition rate that together provide efficient PDT activation and greatly reduced potential for biological damage.

  3. Quantitative comparison of the autofluorescence of bacteria and polystyrene microspheres under violet wavelength excitation for verification of fluorescence-based bioaerosol detector results.

    Science.gov (United States)

    Hasegawa, Norio

    2013-01-01

    The autofluorescence intensity of bacteria and fungal spores was quantified by fluorescence microscopy in order to obtain the information for evaluating fluorescence-based bioaerosol detectors and was comparable to that of some types of polystyrene microspheres (PSMs). Although the intensity for microbes was distributed across a wide range over an order of magnitude in gray scale, it was in the intensity range of certain PSMs. Furthermore, some of those bacteria and PSMs were aerosolized in a test chamber and the fluorescence intensity was measured with a bioaerosol detector. Although there was a slight difference in the order of intensity from the results obtained by fluorescence microscopy, the fluorescence-based bioaerosol detector showed the intensity was in a comparable range.

  4. Upconverting nanoparticles: a versatile platform for wide-field two-photon microscopy and multi-modal in vivo imaging.

    Science.gov (United States)

    Park, Yong Il; Lee, Kang Taek; Suh, Yung Doug; Hyeon, Taeghwan

    2015-03-21

    Lanthanide-doped upconverting nanoparticles (UCNPs) have recently attracted enormous attention in the field of biological imaging owing to their unique optical properties: (1) efficient upconversion photoluminescence, which is intense enough to be detected at the single-particle level with a (nonscanning) wide-field microscope setup equipped with a continuous wave (CW) near-infrared (NIR) laser (980 nm), and (2) resistance to photoblinking and photobleaching. Moreover, the use of NIR excitation minimizes adverse photoinduced effects such as cellular photodamage and the autofluorescence background. Finally, the cytotoxicity of UCNPs is much lower than that of other nanoparticle systems. All these advantages can be exploited simultaneously without any conflicts, which enables the establishment of a novel UCNP-based platform for wide-field two-photon microscopy. UCNPs are also useful for multimodal in vivo imaging because simple variations in the composition of the lattice atoms and dopant ions integrated into the particles can be easily implemented, yielding various distinct biomedical activities relevant to magnetic resonance imaging (MRI), computed tomography (CT), and positron emission tomography (PET). These multiple functions embedded in a single type of UCNPs play a crucial role in precise disease diagnosis. The application of UCNPs is extended to therapeutic fields such as photodynamic and photothermal cancer therapies through advanced surface conjugation schemes.

  5. Two-photon microscopes and in vivo multiphoton tomographs--powerful diagnostic tools for tissue engineering and drug delivery.

    Science.gov (United States)

    Schenke-Layland, Katja; Riemann, Iris; Damour, Odile; Stock, Ulrich A; König, Karsten

    2006-09-15

    Near-infrared multiphoton microscopes and in vivo femtosecond laser tomographs are novel powerful diagnostic tools for intra-tissue drug screening and high-resolution structural imaging applicable to many areas of biomedical research. Deep tissue cells and extracellular matrix (ECM) compartments can be visualized in situ with submicron resolution without the need for tissue processing. In particular, the reduced fluorescent coenzyme NAD(P)H, flavoproteins, keratin, melanin, and elastin are detected by two-photon excited autofluorescence, whereas myosin, tubulin and the ECM protein collagen can be imaged additionally by second harmonic generation (SHG). Therefore, these innovative multiphoton technologies have been used to probe architecture and state of a variety of native tissues, as well as of tissue-engineered constructs, giving insights on the interaction between scaffolds and seeded cells in vitro prior implantation. Moreover, non-invasive 4-D multiphoton tomographs are employed in clinical studies to examine the diffusion behavior, the intra-tissue accumulation of topically applied cosmetic and pharmaceutical components, and their interaction with skin cells.

  6. Two-photon absorption and spectroscopy of the lowest two-photon transition in small donor-acceptor-substituted organic molecules

    Science.gov (United States)

    Beels, Marten T.; Biaggio, Ivan; Reekie, Tristan; Chiu, Melanie; Diederich, François

    2015-04-01

    We determine the dispersion of the third-order polarizability of small donor-acceptor substituted organic molecules using wavelength-dependent degenerate four-wave mixing experiments in solutions with varying concentrations. We find that donor-acceptor-substituted molecules that are characterized by extremely efficient off-resonant nonlinearities also have a correspondingly high two-photon absorption cross section. The width and shape of the first two-photon resonance for these noncentrosymmetric molecules follows what is expected from their longest wavelength absorption peak, and the observed two-photon absorption cross sections are record high when compared to the available literature data, the size of the molecule, and the fundamental limit for two-photon absorption to the lowest excited state, which is essentially determined by the number of conjugated electrons and the excited-state energies. The two-photon absorption of the smallest molecule, which only has 16 electrons in its conjugated system, is one order of magnitude larger than for the molecule called AF-50, a reference molecule for two-photon absorption [O.-K. Kim et al., Chem. Mater. 12, 284 (2000), 10.1021/cm990662r].

  7. Medical prototyping using two photon polymerization

    Directory of Open Access Journals (Sweden)

    Roger J Narayan

    2010-12-01

    Full Text Available Two photon polymerization involves nearly simultaneous absorption of ultrashort laser pulses for selective curing of photosensitive material. This process has recently been used to create small-scale medical devices out of several classes of photosensitive materials, such as acrylate-based polymers, organically-modified ceramic materials, zirconium sol-gels, and titanium-containing hybrid materials. In this review, the use of two photon polymerization for fabrication of several types of small-scale medical devices, including microneedles, artificial tissues, microfluidic devices, pumps, sensors, and valves, from computer models is described. Necessary steps in the development of two photon polymerization as a commercially viable medical device manufacturing method are also considered.

  8. Two Photon Couplings of Hybrid Mesons

    CERN Document Server

    Page, P R

    1996-01-01

    A new formalism is developed for the two photon production of hybrid mesons via intermediate hadronic decays. In an adiabatic and non--relativistic context with spin 1 pair creation we obtain the first absolute estimates of unmixed hybrid production strengths to be small (0.03 - 3 eV) in relation to experimental meson widths (0.1 - 5 keV). Within this context, two photon collisions therefore strongly discriminate between hybrid and conventional meson wave function components at BaBar, Cleo II, LEP2 and LHC, filtering out non--gluonic components. Decay widths of unmixed hybrids are tiny. The formalism also induces conventional meson two photon widths roughly in agreement with experiment.

  9. Two-photon neuronal and astrocytic stimulation with azobenzene-based photoswitches.

    Science.gov (United States)

    Izquierdo-Serra, Mercè; Gascón-Moya, Marta; Hirtz, Jan J; Pittolo, Silvia; Poskanzer, Kira E; Ferrer, Èric; Alibés, Ramon; Busqué, Félix; Yuste, Rafael; Hernando, Jordi; Gorostiza, Pau

    2014-06-18

    Synthetic photochromic compounds can be designed to control a variety of proteins and their biochemical functions in living cells, but the high spatiotemporal precision and tissue penetration of two-photon stimulation have never been investigated in these molecules. Here we demonstrate two-photon excitation of azobenzene-based protein switches and versatile strategies to enhance their photochemical responses. This enables new applications to control the activation of neurons and astrocytes with cellular and subcellular resolution.

  10. Two-photon physics at LEP2

    Energy Technology Data Exchange (ETDEWEB)

    Cartwright, Susan; Lehto, Mark [University of Sheffield Department of Physics, Sheffield S3 7RH (United Kingdom); Seymour, Michael H.; Close, Frank; Wright, Alison [Rutherford Appleton Laboratory, Chilton, Didcot, Oxfordshire OX11 0QX (United Kingdom); Affholderbach, Klaus; Cowan, Glen [Universitaet Siegen, Fachbereich Physik, D-57068 Siegen (Germany); Finch, Alex [University of Lancaster, Lancaster LA1 4YB (United Kingdom); Lauber, Jan [University College London, Gower Street, London WC1E 6BT (United Kingdom)

    1998-02-01

    The working group on two-photon physics concentrated on three main subtopics: modelling the hadronic final state of deep inelastic scattering on a photon; unfolding the deep inelastic scattering data to obtain the photon structure function; and resonant production of exclusive final states, particularly of glueball candidates. In all three areas, new results were presented. (author)

  11. Second harmonic generation and two-photon luminescence upconversion in glasses doped with ZnSe nanocrystalline quantum dots

    Energy Technology Data Exchange (ETDEWEB)

    Thantu, Napoleon [Idaho National Engineering and Environmental Laboratory, 2525 Fremont Avenue, Idaho Falls, ID 83415 (United States)]. E-mail: Napoleon.Thantu@ngc.com

    2005-01-01

    We report two-photon excited emission in borosilicate glasses doped with ZnSe nanocrystalline quantum dots. The emission, predominantly near the two-photon energy and detected in the direction of the excitation beam, is in the visible, and the fundamental excitation is the near-infrared output of a tunable femtosecond laser. Depending on the two-photon energy, time- and frequency-resolved measurements at room temperature reveal that the emission largely consists of second harmonic generation (SHG) and two-photon luminescence upconversion, and a much smaller luminescence from redshifted, low-lying trap states and other trap levels residing near the semiconductor band edge. We discuss the SHG origin in terms of bulk-like and surface contributions from the nanocrystals and the two-photon resonant enhancement near the excitonic absorption.

  12. Skin autofluorescence as a biological UVR dosimeter.

    Science.gov (United States)

    Sandby-Møller, J; Thieden, E; Philipsen, P A; Heydenreich, J; Wulf, H C

    2004-02-01

    Collagen is one of the major endogenous skin fluorophores. Alteration in the structure of collagen due to chronic ultraviolet radiation (UVR) exposure may influence the intensity of the autofluorescence. The aim of this study was to investigate the relation between collagen-linked autofluorescence and sun exposure to clarify whether the skin can be used as a biological UVR dosimeter. We conducted an in vivo study with 131 healthy volunteers. Fluorescence was measured from sun-exposed (dorsal forearm, forehead and shoulder) and sun-protected (buttock) skin and corrected for the impact of pigmentation and redness. The excitation wavelengths (Ex) and emission wavelengths (Em) were: Ex330:Em370, Ex330:Em455 and Ex370:Em455 nm. Individual UVR exposure data were collected both retrospectively and prospectively using questionnaires and electronic personal UVR dosimeters for a summer period. Age, but not sex, skin type or smoking habits correlated significantly positively with skin autofluorescence at Ex370:Em455 at all body sites (P<0.001, r(2)=0.08-0.26), and at Ex330:Em455 only at the buttock (P=0.001, r(2)=0.08), whereas age was not correlated with Ex330:Em370. Sun-protected buttock skin had significantly higher autofluorescence than sun-exposed skin (P-values<0.0001). Because of great between-subject differences in autofluorescence at different body sites, and because the autofluorescence at the unexposed buttock represents the baseline value, individual correction of skin autofluorescence measurement with that of the buttock was performed. Different measures of individual chronic cumulative UVR doses correlated significantly negatively with the skin autofluorescence ratio (F(ratio)), but the correlations were poor (r(2)=0.03-0.10). The results indicate that the collagen-linked skin F(ratio) might be best to use as a measure of individual photodamage, a UVR dose effect, and that it is also a better marker of individual cumulative UVR dose than the used UVR exposure

  13. Spectral characteristics of two-photon autofluorescence and second harmonic generation from human skin in vivo

    Science.gov (United States)

    Breunig, Hans G.; König, Karsten

    2011-03-01

    We performed multiphoton imaging of human skin and recorded in combination the complete spectral content of the signals in vivo. The spectra represent the integration of multiphoton signals over the investigated regions of the epidermis and dermis. They are used to study depth-resolved in vivo emission characteristics of main endogenous skin fluorophores like keratin, NAD(P)H, collagen and elastin. The identification of the specific fluorophores is supported by analysis of additional in vivo fluorescence lifetime imaging. Furthermore, as a potential application of spectrally selective imaging the possibility to investigate the penetration of nanoparticles from sunscreen lotion into skin in vivo is discussed.

  14. Two-Photon Activation of p-Hydroxyphenacyl Phototriggers: Toward Spatially Controlled Release of Diethyl Phosphate and ATP.

    Science.gov (United States)

    Houk, Amanda L; Givens, Richard S; Elles, Christopher G

    2016-03-31

    Two-photon activation of the p-hydroxyphenacyl (pHP) photoactivated protecting group is demonstrated for the first time using visible light at 550 nm from a pulsed laser. Broadband two-photon absorption measurements reveal a strong two-photon transition (>10 GM) near 4.5 eV that closely resembles the lowest-energy band at the same total excitation energy in the one-photon absorption spectrum of the pHP chromophore. The polarization dependence of the two-photon absorption band is consistent with excitation to the same S3 ((1)ππ*) excited state for both one- and two-photon activation. Monitoring the progress of the uncaging reaction under nonresonant excitation at 550 nm confirms a quadratic intensity dependence and that two-photon activation of the uncaging reaction is possible using visible light in the range 500-620 nm. Deprotonation of the pHP chromophore under mildly basic conditions shifts the absorption band to lower energy (3.8 eV) in both the one- and two-photon absorption spectra, suggesting that two-photon activation of the pHP chromophore may be possible using light in the range 550-720 nm. The results of these measurements open the possibility of spatially and temporally selective release of biologically active compounds from the pHP protecting group using visible light from a pulsed laser.

  15. Ocular lens blue autofluorescence cannot be used as a measure of individual cumulative UVR exposure.

    Science.gov (United States)

    Sandby-Møller, Jane; Thieden, Elisabeth; Alshede Philipsen, Peter; Schmidt, Grethe; Wulf, Hans Christian

    2004-02-01

    The accumulation of fluorophores in the ocular lens with age might be caused by ultraviolet solar radiation (UVR) exposure, but evidence of a relation between individual cumulative UVR exposure and lens autofluorescence is lacking. Individually determined UVR exposure has never before been related to lens autofluorescence, and the aim of this study was to investigate if ocular lens blue autofluorescence can be used as a biological UVR dosimeter. Ocular lens autofluorescence was quantified in vivo by fluorescence spectroscopy in 145 volunteers (108 healthy subjects, 18 with basal cell carcinoma (BCC) and 19 with cutaneous malignant melanoma (MM)). The excitation wavelength was 350 nm and the fluorescence emission was 450 nm. Individual UVR exposure data were collected both retrospectively and prospectively using questionnaires and electronic personal UVR dosimeters. Lens blue autofluorescence increased significantly with age (P=0.01), and females had significantly higher autofluorescence than males (P=0.024); the two factors explained 10% of the total variation in lens autofluorescence. Neither smoking habits nor use of glasses/contact lenses or sunglasses influenced autofluorescence. No correlations between autofluorescence and UVR exposure measurements were found, and neither was there a difference in autofluorescence between groups with high and low UVR exposure (P-values>0.1), respectively. MM patients had significantly (P=0.019) higher autofluorescence than healthy subjects when age and sex differences were taken into account; no such difference (P=0.097) was detected between BCC patients and healthy subjects. The results indicate that age and gender only play a minor role in the level of lens blue autofluorescence. Exposure to UVR has been suggested to be responsible for a part of the age-related increase in autofluorescence, but this could not be confirmed in this study. The higher level of lens autofluorescence found in MM patients might be due to genetics

  16. Transparency induced by two photon interference in a beam splitter

    Institute of Scientific and Technical Information of China (English)

    Wang Kai-Ge; Yang Guo-Jian

    2004-01-01

    We propose a special two-photon state which is completely transparent in a 50/50 beam splitter. This effect is caused by the destructive two-photon interference and shows the signature of photon entanglement. We find that the symmetry of the two-photon spectrum plays the key role for the properties of two-photon interference.

  17. Two-photon cooling of magnesium atoms

    DEFF Research Database (Denmark)

    Malossi, N.; Damkjær, S.; Hansen, P. L.

    2005-01-01

    A two-photon mechanism for cooling atoms below the Doppler temperature is analyzed. We consider the magnesium ladder system (3s2)S01¿(3s3p)P11 at 285.2nm followed by the (3s3p)P11¿(3s3d)D21 transition at 880.7nm . For the ladder system quantum coherence effects may become important. Combined...... with the basic two-level Doppler cooling process this allows for reduction of the atomic sample temperature by more than a factor of 10 over a broad frequency range. First experimental evidence for the two-photon cooling process is presented and compared to model calculations. Agreement between theory...... and experiment is excellent. In addition, by properly choosing the Rabi frequencies of the two optical transitions a velocity independent atomic dark state is observed....

  18. Anomalous two-photon spectral features in warm rubidium vapor

    Science.gov (United States)

    Perrella, C.; Light, P. S.; Milburn, T. J.; Kielpinski, D.; Stace, T. M.; Luiten, A. N.

    2016-09-01

    We report observation of anomalous fluorescence spectral features in the environs of a two-photon transition in a rubidium vapor when excited with two different wavelength lasers that are both counterpropagating through the vapor. These features are characterized by an unusual trade-off between the detunings of the driving fields. Three different hypothetical processes are presented to explain the observed spectra: a simultaneous three-atom and four-photon collision, a four-photon excitation involving a light field produced via amplified spontaneous emission, and population pumping perturbing the expected steady-state spectra. Numerical modeling of each hypothetical process is presented, supporting the population pumping process as the most plausible mechanism.

  19. Two-photon ionization of colliding atoms

    Energy Technology Data Exchange (ETDEWEB)

    Nayfeh, M.H.

    1977-09-01

    Semiclassical expressions of two-photon ionization of two colliding atoms are derived for a wide range of electromagnetic field intensity and detunings from the isolated atom line. The dependence of the ionization yield on the details of the interaction potential of the system is derived. This process promises an extremely sensitive method for studying line broadening on the far wing, especially when absorption or fluorescence becomes very weak.

  20. Two-photon cooling of magnesium atoms

    DEFF Research Database (Denmark)

    Malossi, N.; Damkjær, S.; Hansen, P. L.;

    2005-01-01

    A two-photon mechanism for cooling atoms below the Doppler temperature is analyzed. We consider the magnesium ladder system (3s2)S01¿(3s3p)P11 at 285.2nm followed by the (3s3p)P11¿(3s3d)D21 transition at 880.7nm . For the ladder system quantum coherence effects may become important. Combined...

  1. Two-photon super bunching of thermal light via multiple two-photon-path interference

    CERN Document Server

    Hong, Peilong; Zhang, Guoquan

    2012-01-01

    We propose a novel scheme to achieve two-photon super bunching of thermal light through multiple two-photon-path interference, in which two mutually first-order incoherent optical channels are introduced by inserting a modified Michelson interferometer into a traditional two-photon HBT interferometer, and the bunching peak-to-background ratio can reach 3 theoretically. Experimentally, the super bunching peak-to-background ratio was measured to be 2.4, much larger than the ratio 1.7 measured with the same thermal source in a traditional HBT interferometer. The peak-to-background ratio of two-photon super bunching of thermal light can be increased up to $2\\times1.5^n$ by inserting cascadingly $n$ pairs of mutually first-order incoherent optical channels into the traditional two-photon HBT interferometer. The two-photon super bunching of thermal light should be of great significance in improving the visibility of classical ghost imaging.

  2. Observation of Nondegenerate Two-Photon Gain in GaAs

    CERN Document Server

    Reichert, Matthew; Salamo, Greg; Hagan, David J; Van Stryland, Eric W

    2016-01-01

    Two-photon lasers require materials with large two-photon gain (2PG) coefficients and low linear and nonlinear losses. Our previous demonstration of large enhancement of two-photon absorption in semiconductors for very different photon energies translates directly into enhancement of 2PG. We experimentally demonstrate nondegenerate 2PG in optically excited bulk GaAs via femtosecond pump-probe measurements. 2PG is isolated from other pump induced effects through the difference between measurements performed with parallel and perpendicular polarizations of pump and probe. An enhancement in the 2PG coefficient of nearly two orders-of-magnitude is reported. The results point a possible way toward two-photon semiconductor lasers.

  3. One- and two-photon scattering from generalized V-type atoms

    OpenAIRE

    Sánchez-Burillo, Eduardo; Martín-Moreno, Luis; Zueco, David; García-Ripoll, Juan José

    2016-01-01

    The one- and two-photon scattering matrix S is obtained analytically for a one-dimensional waveguide and a point-like scatterer with N excited levels (generalized V -type atom). We argue that the two-photon scattering matrix contains sufficient information to distinguish between different level structures which are equivalent for single-photon scattering, such as a V -atom with N = 2 excited levels and two two-level systems. In particular, we show that the scattering with the V -type atom exh...

  4. Manipulation of multiple electromagnetically induced two-photon transparency in a six-level atomic system

    Institute of Scientific and Technical Information of China (English)

    Jia Wen-Zhi; Wang Shun-Jin

    2009-01-01

    In the five-level K-type atomic system, by using another control field to couple the excited level of the coupling transition to the sixth higher excited level, a six-level atomic system is constructed. In this system, the multiple electromagnetically induced two-photon transparency has been investigated. What is more, if choosing the parameters of the control fields properly the triple transparency window will reduce to a double one which means that the multiple electromagnetically induced two-photon transparency can be manipulated in this system. The physical interpretation of these phenomena is given in terms of the dressed states and the dark states.

  5. Two-photon laser fabrication of three-dimensional silver microstructures with submicron scale linewidth

    Energy Technology Data Exchange (ETDEWEB)

    Tsutsumi, Naoto; Nagata, Kazuya; Sakai, Wataru [Kyoto Institute of Technology, Department of Macromolecular Science and Engineering, Graduate School of Science and Technology, Kyoto (Japan)

    2011-05-15

    We show three-dimensional silver microstructures with a submicron scale linewidth fabricated via two-photon photoreduction of silver ions in a poly(N-vinylpyrrolidone) (PVP) matrix. Femtosecond laser at 508 nm directly excites the carbonyl group of PVP via two-photon excitation to reduce silver ions. Lone pair electrons in PVP stabilized silver ions and lower molecular weight of PVP prevented silver clusters growing larger. The effect of molecular weight of PVP on linewidth of silver nanowire is investigated. (orig.)

  6. One and Two Photon Excitation of Radiofrequency Trapped Ca+

    Institute of Scientific and Technical Information of China (English)

    C. Zumsteg; C. Champenois; D. Guyomarc'h; G. Hagel; M. Houssin; M. Knoop

    2009-01-01

    Radiofrequency (rf) trapped ions are versatile candidates for a large panel of applications ranging from quantum information to the creation of cold molecules. Sample size can range from a single to 106 ions, and the internal and external energy states of the atoms can be controlled with high precision. In the experiment, we focus on different protocols related to frequency metrology using rf trapped Ca+.

  7. Adiabatic following in two-photon transition

    Energy Technology Data Exchange (ETDEWEB)

    Nayfeh, M.H.; Nayfeh, A.H.

    1977-03-01

    The coherent interaction of two smoothly varying, near-resonant, two-photon pulses with a three-level system can be described by ''two-photon damped Bloch equations'' which are analogous to those for a one-photon transition in a two-level system except for the presence of a two-photon coupling and a frequency shift. These equations are solved for the cases ..gamma../sub 1/, ..gamma../sub 2/ very-much-less-than ..cap omega.., ..gamma../sub 1/ = ..gamma../sub 2/, and ..gamma../sub 2/k/sup 2/epsilon/sup 4//..cap omega../sup 2/, ..gamma../sub 1/ very-much-less-than ..cap omega.., where ..gamma../sub 1/ and ..gamma../sub 2/ are the atomic energy and phase relaxation widths, respectively, and ..cap omega.. is the Rabi frequency. The leading contribution to the refractive index is intensity dependent, caused by the level shifts inherent in multiphoton processes; it includes a relaxation dependent part which is important at times shorter than ..gamma../sup -1//sub 1/. The second-order contributions depend on the square of the intensity and the time-integrated square of the intensity. The latter contribution, which is relaxation dependent, causes line asymmetry at the long-wavelength wing; it consists of a term proportional to ..gamma../sub 2/-..gamma../sub 1/ and only important at early times and a term proportional to 2..gamma../sub 2/-..gamma../sub 1/.

  8. Denoising two-photon calcium imaging data.

    Science.gov (United States)

    Malik, Wasim Q; Schummers, James; Sur, Mriganka; Brown, Emery N

    2011-01-01

    Two-photon calcium imaging is now an important tool for in vivo imaging of biological systems. By enabling neuronal population imaging with subcellular resolution, this modality offers an approach for gaining a fundamental understanding of brain anatomy and physiology. Proper analysis of calcium imaging data requires denoising, that is separating the signal from complex physiological noise. To analyze two-photon brain imaging data, we present a signal plus colored noise model in which the signal is represented as harmonic regression and the correlated noise is represented as an order autoregressive process. We provide an efficient cyclic descent algorithm to compute approximate maximum likelihood parameter estimates by combing a weighted least-squares procedure with the Burg algorithm. We use Akaike information criterion to guide selection of the harmonic regression and the autoregressive model orders. Our flexible yet parsimonious modeling approach reliably separates stimulus-evoked fluorescence response from background activity and noise, assesses goodness of fit, and estimates confidence intervals and signal-to-noise ratio. This refined separation leads to appreciably enhanced image contrast for individual cells including clear delineation of subcellular details and network activity. The application of our approach to in vivo imaging data recorded in the ferret primary visual cortex demonstrates that our method yields substantially denoised signal estimates. We also provide a general Volterra series framework for deriving this and other signal plus correlated noise models for imaging. This approach to analyzing two-photon calcium imaging data may be readily adapted to other computational biology problems which apply correlated noise models.

  9. Biological oxygen sensing via two-photon absorption by an Ir(III) complex using a femtosecond fiber laser

    Science.gov (United States)

    Moritomo, Hiroki; Fujii, Akinari; Suzuki, Yasutaka; Yoshihara, Toshitada; Tobita, Seiji; Kawamata, Jun

    2016-09-01

    Near-infrared two-photon absorption of the phosphorescent Ir(III) complex (2,4-pentanedionato-κO 2,κO 4)bis[2-(6-phenanthridinyl-κN)benzo[b]thien-3-yl-κC]iridium (BTPHSA) was characterized. It exhibited a 800-1200 nm two-photon absorption band, and thus could be electronically excited by 1030-nm femtosecond Ti:sapphire and Yb-doped fiber lasers. By using BTPHSA, oxygen concentrations in human embryonic kidney 293 (HEK293) cells were imaged. These results demonstrate two-photon oxygen sensing of live tissues via easily operable excitation sources.

  10. Enhanced-locality fiber-optic two-photon-fluorescence live-brain interrogation

    Energy Technology Data Exchange (ETDEWEB)

    Fedotov, I. V.; Doronina-Amitonova, L. V. [International Laser Center, Physics Department, M.V. Lomonosov Moscow State University, Moscow 119992 (Russian Federation); Russian Quantum Center, ul. Novaya 100, Skolkovo, Moscow Region 1430125 (Russian Federation); Kurchatov Institute National Research Center, Moscow (Russian Federation); Sidorov-Biryukov, D. A.; Fedotov, A. B. [International Laser Center, Physics Department, M.V. Lomonosov Moscow State University, Moscow 119992 (Russian Federation); Russian Quantum Center, ul. Novaya 100, Skolkovo, Moscow Region 1430125 (Russian Federation); Anokhin, K. V. [Kurchatov Institute National Research Center, Moscow (Russian Federation); P.K. Anokhin Institute of Normal Physiology, Russian Academy of Medical Sciences, Moscow (Russian Federation); Kilin, S. Ya. [B.I. Stepanov Institute of Physics, National Academy of Sciences of Belarus, Minsk (Belarus); Sakoda, K. [National Institute for Materials Science, 1-1 Namiki, Tsukuba 305-0044 (Japan); Zheltikov, A. M. [International Laser Center, Physics Department, M.V. Lomonosov Moscow State University, Moscow 119992 (Russian Federation); Russian Quantum Center, ul. Novaya 100, Skolkovo, Moscow Region 1430125 (Russian Federation); Department of Physics and Astronomy, Texas A and M University, College Station, Texas 77843 (United States); Center of Photochemistry, Russian Academy of Sciences, ul. Novatorov 7a, Moscow 117421 (Russian Federation)

    2014-02-24

    Two-photon excitation is shown to substantially enhance the locality of fiber-based optical interrogation of strongly scattering biotissues. In our experiments, a high-numerical-aperture, large-core-are fiber probe is used to deliver the 200-fs output of a 100-MHz mode-locked ytterbium fiber laser to samples of live mouse brain, induce two-photon fluorescence of nitrogen–vacancy centers in diamond markers in brain sample. Fiber probes with a high numerical aperture and a large core area are shown to enable locality enhancement in fiber-laser–fiber-probe two-photon brain excitation and interrogation without sacrificing the efficiency of fluorescence response collection.

  11. Polarization properties of optical phase conjugation by two-photon resonant degenerate four-wave mixing

    Science.gov (United States)

    Kauranen, Martti; Gauthier, Daniel J.; Malcuit, Michelle S.; Boyd, Robert W.

    1989-08-01

    We develop a semiclassical theory of the polarization properties of phase conjugation by two-photon resonant degenerate four-wave mixing. The theory includes the effects of saturation by the pump waves. We solve the density-matrix equations of motion in steady state for a nonlinear medium consisting of stationary atoms with a ground and excited state connected by two-photon transitions. As an illustration of the general results, we consider an S0-->S0 two-photon transition, which is known to lead to perfect polarization conjugation in the limit of third-order theory. We show that the fidelity of the polarization-conjugation process is degraded for excessively large pump intensities. The degradation can occur both due to transfer of population to the excited state and due to nonresonant Stark shifts. Theoretical results are compared to those of a recent experiment [Malcuit, Gauthier, and Boyd, Opt. Lett. 13, 663 (1988)].

  12. A spirobifluorene-based two-photon fluorescence probe for mercury ions and its applications in living cells

    Energy Technology Data Exchange (ETDEWEB)

    Xiao, Haibo, E-mail: xiaohb@shnu.edu.cn; Zhang, Yanzhen; Zhang, Wu; Li, Shaozhi; Tan, Jingjing; Han, Zhongying

    2017-05-01

    A novel spirobifluorene derivative SPF-TMS, which containing dithioacetal groups and triphenylamine units, was synthesized. The probing behaviors toward various metal ions were investigated via UV/Vis absorption spectra as well as one-photon fluorescence changes. The results indicated that SPF-TMS exhibits high sensitivity and selectivity for mercury ions. The detection limit was at least 8.6 × 10{sup −8}M, which is excellent comparing with other optical sensors for Hg{sup 2+}. When measured by two-photon excited fluorescence technique in THF at 800 nm, the two-photon cross-section of SPF-TMS is 272 GM. Especially, upon reaction with mercury species, SPF-TMS yielded another two-photon dye SPF-DA. Both SPF-TMS and SPF-DA emit strong two-photon induced fluorescence and can be applied in cell imaging by two-photon microscopy. - Highlights: • We report a spirobifluorene-based molecule as two-photon fluorescent probe with large two-photon cross-section. • The molecule has exclusive selectivity and sensitivity for mercury species. • The molecule has large two-photon emission changes before and after addition of Hg{sup 2+}. • Both the probe and the mercury ion-promoted reaction product can be applied in cell imaging by two-photon microscopy.

  13. Two-Photon and Second Harmonic Microscopy in Clinical and Translational Cancer Research

    Science.gov (United States)

    PERRY, SETH W.; BURKE, RYAN M.; BROWN, EDWARD B.

    2012-01-01

    Application of two-photon microscopy (TPM) to translational and clinical cancer research has burgeoned over the last several years, as several avenues of pre-clinical research have come to fruition. In this review, we focus on two forms of TPM—two-photon excitation fluorescence microscopy, and second harmonic generation microscopy—as they have been used for investigating cancer pathology in ex vivo and in vivo human tissue. We begin with discussion of two-photon theory and instrumentation particularly as applicable to cancer research, followed by an overview of some of the relevant cancer research literature in areas that include two-photon imaging of human tissue biopsies, human skin in vivo, and the rapidly developing technology of two-photon microendoscopy. We believe these and other evolving two-photon methodologies will continue to help translate cancer research from the bench to the bedside, and ultimately bring minimally invasive methods for cancer diagnosis and treatment to therapeutic reality. PMID:22258888

  14. Two-photon resonant, stimulated processes in krypton and xenon

    Energy Technology Data Exchange (ETDEWEB)

    Miller, J.C.

    1988-11-01

    Both on-axis and conical emissions have been observed following two-photon pumping of the 5p states of krypton and the 6p', 7p, 8p, and 4f states of xenon. In the former case, coherent emissions from the 5p states to the 5s are observed, and in the latter case, many p..-->..s, d..-->..p, and f..-->..d cascade emissions are observed. By analogy to the well-studied alkali and alkaline earth examples, the emissions are discussed in terms of amplified spontaneous emission (ASE), stimulated hyper-Raman scattering, and parametric four-wave mixing. The physical processes responsible for the conical emission and for intensity anomalies in the xenon p..-->..s emissions are not understood at present. Interference effects due to coherent cancellation between competing excitation pathways may be occurring. 4 refs., 3 figs.

  15. Nuclear two-photon decay in 0 +→0 + transitions

    Science.gov (United States)

    Kramp, J.; Habs, D.; Kroth, R.; Music, M.; Schirmer, J.; Schwalm, D.; Broude, C.

    1987-11-01

    The two-photon decay of the first excited 0 + state of 16O has been measured using the Heidelberg-Darmstadt crystal ball. A branching ratio of {Γ γγ}/{Γ tot} = (6.6±0.5) · 10 -4 was obtained. As in the cases of 40Ca and 90Zr previously reported by us, the 2γ decay of 16O proceeds via double E1 and M1 transitions of similar strength; the evidence is the observed interference term in the 2γ angular correlation. The ratio of the matrix elements {α E1 }/{χ} for 16O was restricted to the two inverse values (-6.2±1.5) or (-0.16±0.04). An interpretation of 2γ matrix elements observed for 16O, 40Ca and 90Zr in terms of the electric polarizabilities and magnetic susceptibility is given leading to a qualitative understanding of this decay mode.

  16. Investigate the variation in optical redox ratio of epicardial adipose tissue in patients with CAD through auto-fluorescence metabolic molecular image (Conference Presentation)

    Science.gov (United States)

    Guo, Lun-Zhang; Wang, Tzung-Dau; Lin, Jong-Wei; Liu, Tzu-Ming

    2016-04-01

    In recent years, it has been suggested that epicardial adipose tissue (EAT) plays an important role in development of coronary artery disease (CAD) and diabetes mellitus (DM). In this article, we used two-photon fluoresce microscope to measure the fluorescence metabolic image of EAT, which obtained from the patient with/without CAD/DM. We used 740nm and 890nm infrared light to excite the auto-fluorescence of metabolic molecules NADH and FAD respectively. We collected the fluorescence signal at wavelength 450nm to 500nm and 500nm to 550nm to obtain the metabolic image. Through the image, we computed the redox ratio (NADH/FAD) by analyzing the intensity. The preliminary result showed that the redox ratio increase in the patients with CAD. It indicates EAT adipocytes of patient with CAD have decreased cellular metabolic activity. But there were no significant variation of redox ratio in the patients with DM.

  17. Carbon quantum dot-NO photoreleaser nanohybrids for two-photon phototherapy of hypoxic tumors.

    Science.gov (United States)

    Fowley, Colin; McHale, Anthony P; McCaughan, Bridgeen; Fraix, Aurore; Sortino, Salvatore; Callan, John F

    2015-01-04

    We report a conjugate between carbon quantum dots and a NO photoreleaser able to photogenerate the anticancer NO radical via an energy transfer mechanism. This nanohybrid proved toxic to cancer cells in vitro and significantly reduced tumor volume in mice bearing human xenograft BxPC-3 pancreatic tumors upon two-photon excitation with the highly biocompatible 800 nm light.

  18. Probing Electron-Phonon Interaction through Two-Photon Interference in Resonantly Driven Semiconductor Quantum Dots

    DEFF Research Database (Denmark)

    Reigue, Antoine; Iles-Smith, Jake; Lux, Fabian

    2017-01-01

    We investigate the temperature dependence of photon coherence properties through two-photon interference (TPI) measurements from a single quantum dot (QD) under resonant excitation. We show that the loss of indistinguishability is related only to the electron-phonon coupling and is not affected...

  19. Two-photon interference : spatial aspects of two-photon entanglement, diffraction, and scattering

    NARCIS (Netherlands)

    Peeters, Wouter Herman

    2010-01-01

    This dissertation contains scientific research within the realm of quantum optics, which is a branch of physics. An experimental and theoretical study is made of two-photon interference phenomena in various optical systems. Spatially entangled photon pairs are produced via the nonlinear optical proc

  20. Integrating two-photon microscopy and cryo-electron microscopy for studying the interaction of Cafeteria roenbergensis and CroV

    Science.gov (United States)

    Aghvami, Seyedmohammadali

    Cafeteria roenbergensis (Cro) is a marine zooplankton; its voracious appetite plays a significant role in regulating bacteria populations. The giant virus that lives within Cro, known as Cafeteria roenbergensis virus (CroV), has an important effect on the mortality of Cro populations. Although viral infections are extremely abundant in oceans, the complete procedure of the infection is still unknown. We study the infection process of Cro by CroV to find out whether the initial contact is through phagocytosis or CroV penetrating the host cell membrane directly. Cro is a moving at speed in the range of 10-100 um/s, therefore, there are many difficulties and challenges for traditional imaging techniques to study this viral-host interaction. We apply two-photon fluorescence microscopy to image this infection process. The image is taken at video rate (30 frame/s), which makes us able to catch the moment of interaction. We are able to image host and virus simultaneously where CroV is stained by SYBR gold dye and Cro is excited through NADH autofluorescence. For further structural biology study, we will obtain atomic level resolution information of infection. After catching the initial moment of infection, we will freeze the sample instantly and image it with cryo-electron microscope .

  1. Spectral distribution of the 2 → 1 two-photon transition in atoms and few-electron ions

    Indian Academy of Sciences (India)

    Ajay Kumar; S Trotsenko; A V Volotka; D Banaś; H F Beyer; H Bräuning; S Fritzsche; A Gumberidze; S Hagmann; S Hess; C Kozhuharov; R Reuschl; U Spillmann; M Trassinelli; G Weber; Th Stöhlker

    2011-02-01

    The two-photon decay of the 2 state to the ground state in dressed atoms and oneor two-electron ions has been studied for several decades. Relativistic calculations have shown an -dependence of the spectral shape of this two-photon transition in one- or two-electron ions. We have measured the spectral distribution of the 121 0 → 12 1 0 two-photon transition in He-like tin at the ESR storage ring using a new approach for such experiments. In this method, relativistic collisions of initially Li-like projectiles with a gaseous target were used to populate exclusively the first excited state, 12, of He-like tin, which provided a clean two-photon spectrum. The measured two-photon spectral distribution was compared with fully relativistic calculations. The obtained results show very good agreement with the calculations for He-like tin

  2. Two-photon Interference with Non-identical Photons

    CERN Document Server

    Liu, Jianbin; Zheng, Huaibin; Chen, Hui; Li, Fu-Li; Xu, Zhuo

    2014-01-01

    The indistinguishability of non-identical photons is dependent on detection system in quantum physics. If two photons with different wavelengths are indistinguishable for a detection system, there can be two-photon interference when these two photons are incident to two input ports of a Hong-Ou-Mandel interferometer, respectively. The reason why two-photon interference phenomena are different for classical and nonclassical light is not due to interference, but due to the properties of light and detection system. These conclusions are helpful to understand the physics and applications of two-photon interference.

  3. Insights into esophagus tissue architecture using two-photon confocal microscopy

    Science.gov (United States)

    Liu, Nenrong; Wang, Yue; Feng, Shangyuan; Chen, Rong

    2013-08-01

    In this paper, microstructures of human esophageal mucosa were evaluated using the two-photon laser scanning confocal microscopy (TPLSCM), based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG). The distribution of epithelial cells, muscle fibers of muscularis mucosae has been distinctly obtained. Furthermore, esophageal submucosa characteristics with cancer cells invading into were detected. The variation of collagen, elastin and cancer cells is very relevant to the pathology in esophagus, especially early esophageal cancer. Our experimental results indicate that the MPM technique has the much more advantages for label-free imaging, and has the potential application in vivo in the clinical diagnosis and monitoring of early esophageal cancer.

  4. Near IR two photon absorption of cyanines dyes: application to optical power limiting at telecommunication wavelengths

    Science.gov (United States)

    Bouit, Pierre-Antoine; Wetzel, Guillaume; Feneyrou, Patrick; Bretonnière, Yann; Kamada, Kenji; Maury, Olivier; Andraud, Chantal

    2008-02-01

    The design and synthesis of symmetrical and unsymmetrical heptamethine cyanines is reported. These chromophores present significant two-photon cross section in the 1400-1600 nm spectral range. In addition, they display optical power limiting (OPL) properties. OPL curves were interpreted on the basis of two-photon absorption (2PA) followed by excited state absorption (ESA). Finally, these molecules present several relevant properties (nonlinear absorption properties, two-step gram scale synthesis, high solubility, good thermal stability), which could lead to numerous practical applications in material science (solid state optical limiting, signal processing) or in biology (imaging).

  5. In-vivo two-photon imaging of the honey bee antennal lobe

    CERN Document Server

    Haase, Albrecht; Trona, Federica; Anfora, Gianfranco; Vallortigara, Giorgio; Antolini, Renzo; Vinegoni, Claudio

    2010-01-01

    Due to the honey bee's importance as a simple neural model, there is a great need for new functional imaging modalities. Herein we report on the use of two-photon microscopy for in-vivo functional and morphological imaging of the honey bee's olfactory system focusing on its primary centers, the antennal lobes (ALs). Our imaging platform allows for simultaneously obtaining both morphological measurements of the AL and in-vivo calcium recording of neural activities. By applying external odor stimuli to the bee's antennas, we were able to record the characteristic odor response maps. Compared to previous works where conventional fluorescence microscopy is used, our approach offers all the typical advantages of multi-photon imaging, providing substantial enhancement in both spatial and temporal resolutions while minimizing photo-damages and autofluorescence contribution with a four-fold improvement in the functional signal. Moreover, the multi-photon associated extended penetration depth allows for functional ima...

  6. Fully integrated reflection-mode photoacoustic/two-photon microscopy in vivo (Conference Presentation)

    Science.gov (United States)

    Song, Liang; Song, Wei; Zhang, Yang; Zheng, Wei

    2016-03-01

    Using a water-immersion optical objective in conjunction with a miniature 40-MHz ultrasonic transducer, we developed reflection-mode photoacoustic microscopy with a transverse resolution as high as 320 nm. Here, we further integrated two-photon microscopy capability into the system to enable multimodality in vivo biomedical imaging at submicron resolution. As a result, the system is capable of tri-modality label-free imaging of microvasculature, collagen, and cell morphology, based on the contrast of optical absorption, second-harmonic generation, and autofluorescence, respectively. In addition, we demonstrated simultaneous microscopic imaging of neuron and microvasculature in the brain cortex of a living mouse, which may offer new opportunities for studying the mechanisms of neurovascular coupling.

  7. Mitigating thermal mechanical damage potential during two-photon dermal imaging.

    Science.gov (United States)

    Masters, Barry R; So, Peter T C; Buehler, Christof; Barry, Nicholas; Sutin, Jason D; Mantulin, William W; Gratton, Enrico

    2004-01-01

    Two-photon excitation fluorescence microscopy allows in vivo high-resolution imaging of human skin structure and biochemistry with a penetration depth over 100 microm. The major damage mechanism during two-photon skin imaging is associated with the formation of cavitation at the epidermal-dermal junction, which results in thermal mechanical damage of the tissue. In this report, we verify that this damage mechanism is of thermal origin and is associated with one-photon absorption of infrared excitation light by melanin granules present in the epidermal-dermal junction. The thermal mechanical damage threshold for selected Caucasian skin specimens from a skin bank as a function of laser pulse energy and repetition rate has been determined. The experimentally established thermal mechanical damage threshold is consistent with a simple heat diffusion model for skin under femtosecond pulse laser illumination. Minimizing thermal mechanical damage is vital for the potential use of two-photon imaging in noninvasive optical biopsy of human skin in vivo. We describe a technique to mitigate specimen thermal mechanical damage based on the use of a laser pulse picker that reduces the laser repetition rate by selecting a fraction of pulses from a laser pulse train. Since the laser pulse picker decreases laser average power while maintaining laser pulse peak power, thermal mechanical damage can be minimized while two-photon fluorescence excitation efficiency is maximized.

  8. RuBi-Glutamate: Two-Photon and Visible-Light Photoactivation of Neurons and Dendritic spines.

    Science.gov (United States)

    Fino, Elodie; Araya, Roberto; Peterka, Darcy S; Salierno, Marcelo; Etchenique, Roberto; Yuste, Rafael

    2009-01-01

    We describe neurobiological applications of RuBi-Glutamate, a novel caged-glutamate compound based on ruthenium photochemistry. RuBi-Glutamate can be excited with visible wavelengths and releases glutamate after one- or two-photon excitation. It has high quantum efficiency and can be used at low concentrations, partly avoiding the blockade of GABAergic transmission present with other caged compounds. Two-photon uncaging of RuBi-Glutamate has a high spatial resolution and generates excitatory responses in individual dendritic spines with physiological kinetics. With laser beam multiplexing, two-photon RuBi-Glutamate uncaging can also be used to depolarize and fire pyramidal neurons with single-cell resolution. RuBi-Glutamate therefore enables the photoactivation of neuronal dendrites and circuits with visible or two-photon light sources, achieving single cell, or even single spine, precision.

  9. Visualization of two-photon Rabi oscillations in evanescently coupled optical waveguides

    Energy Technology Data Exchange (ETDEWEB)

    Ornigotti, M; Valle, G Della; Fernandez, T Toney; Laporta, P; Longhi, S [Dipartimento di Fisica and Istituto di Fotonica e Nanotecnologie del CNR, Politecnico di Milano, Piazza L. da Vinci 32, I-20133 Milano (Italy); Coppa, A; Foglietti, V [Istituto di Fotonica e Nanotecnologie del CNR, sezione di Roma, Via Cineto Romano 42, 00156 Roma (Italy)], E-mail: longhi@fisi.polimi.it

    2008-04-28

    An optical analogue of two-photon Rabi oscillations, occurring in a three-level atomic or molecular system coherently driven by two detuned laser fields, is theoretically proposed and experimentally demonstrated using three evanescently coupled optical waveguides realized on an active glass substrate. The optical analogue stems from the formal analogy between spatial propagation of light waves in the three-waveguide structure and the coherent temporal evolution of populations in a three-level atomic medium driven by two laser fields under two-photon resonance. In our optical experiment, two-photon Rabi oscillations are thus visualized as a slow spatial oscillatory exchange of light power between the two outer waveguides of the structure with a small excitation of the central waveguide.

  10. Three-dimensional protein networks assembled by two-photon activation.

    Science.gov (United States)

    Gatterdam, Volker; Ramadass, Radhan; Stoess, Tatjana; Fichte, Manuela A H; Wachtveitl, Josef; Heckel, Alexander; Tampé, Robert

    2014-05-26

    Spatial and temporal control over chemical and biological processes plays a key role in life and material sciences. Here we synthesized a two-photon-activatable glutathione (GSH) to trigger the interaction with glutathione S-transferase (GST) by light at superior spatiotemporal resolution. The compound shows fast and well-confined photoconversion into the bioactive GSH, which is free to interact with GST-tagged proteins. The GSH/GST interaction can be phototriggered, changing its affinity over several orders of magnitude into the nanomolar range. Multiplexed three-dimensional (3D) protein networks are simultaneously generated in situ through two-photon fs-pulsed laser-scanning excitation. The two-photon activation facilitates the three-dimensional assembly of protein structures in real time at hitherto unseen resolution in time and space, thus opening up new applications far beyond the presented examples.

  11. Two-Photon Absorption Properties of Mn-Doped ZnS Quantum Dots

    Institute of Scientific and Technical Information of China (English)

    ZHENG Jia-Jin; ZHANG Gui-Lan; GUO Yang-Xue; WANG Xiao-Yan; CHEN Wen-Ju; ZHANG Xiao-Song; HUA Yu-Lin

    2006-01-01

    @@ We investigate the two-photon absorption and nonlinear refractive index properties of a quantum dot material based on ZnS nanocrystals doped with Mn isoelectronic impurities, using the Z-scan technique with 532nm picosecond laser pulses. The Mn-doped ZnS quantum dots have an average two-photon absorption cross section as high as 13600 Goeppert-Mayer units, which turn it into a very promising material for fluorescent label and imaging in biological samples. In addition, we also found that the two-photon absorption coeflicient initially increases and then decreases with increasing pulse irradiance, which demonstrates the presence of the higherorder nonlinearity under the strong excitation.

  12. Scanless functional imaging of hippocampal networks using patterned two-photon illumination through GRIN lenses

    KAUST Repository

    Moretti, Claudio

    2016-09-12

    Patterned illumination through the phase modulation of light is increasingly recognized as a powerful tool to investigate biological tissues in combination with two-photon excitation and light-sensitive molecules. However, to date two-photon patterned illumination has only been coupled to traditional microscope objectives, thus limiting the applicability of these methods to superficial biological structures. Here, we show that phase modulation can be used to efficiently project complex two-photon light patterns, including arrays of points and large shapes, in the focal plane of graded index (GRIN) lenses. Moreover, using this approach in combination with the genetically encoded calcium indicator GCaMP6, we validate our system performing scanless functional imaging in rodent hippocampal networks in vivo ~1.2 mm below the brain surface. Our results open the way to the application of patterned illumination approaches to deep regions of highly scattering biological tissues, such as the mammalian brain.

  13. Two-Photon Absorption in Conjugated Energetic Molecules.

    Science.gov (United States)

    Bjorgaard, Josiah A; Sifain, Andrew E; Nelson, Tammie; Myers, Thomas W; Veauthier, Jacqueline M; Chavez, David E; Scharff, R Jason; Tretiak, Sergei

    2016-07-07

    Time-dependent density functional theory (TD-DFT) was used to investigate the relationship between molecular structure and the one- and two-photon absorption (OPA and TPA, respectively) properties of novel and recently synthesized conjugated energetic molecules (CEMs). The molecular structures of CEMs can be strategically altered to influence the heat of formation and oxygen balance, two factors that can contribute to the sensitivity and strength of an explosive material. OPA and TPA are sensitive to changes in molecular structure as well, influencing the optical range of excitation. We found calculated vertical excitation energies to be in good agreement with experiment for most molecules. Peak TPA intensities were found to be significant and on the order of 10(2) GM. Natural transition orbitals for essential electronic states defining TPA peaks of relatively large intensity were used to examine the character of relevant transitions. Modification of molecular substituents, such as additional oxygen or other functional groups, produces significant changes in electronic structure, OPA, and TPA and improves oxygen balance. The results show that certain molecules are apt to undergo nonlinear absorption, opening the possibility for controlled, direct optical initiation of CEMs through photochemical pathways.

  14. Synergistic Two-Photon Absorption Enhancement in Photosynthetic Light Harvesting

    Science.gov (United States)

    Chen, Kuo-Mei; Chen, Yu-Wei; Gao, Ting-Fong

    2012-06-01

    The grand scale fixation of solar energies into chemical substances by photosynthetic reactions of light-harvesting organisms provides Earth's other life forms a thriving environment. Scientific explorations in the past decades have unraveled the fundamental photophysical and photochemical processes in photosynthesis. Higher plants, green algae, and light-harvesting bacteria utilize organized pigment-protein complexes to harvest solar power efficiently and the resultant electronic excitations are funneled into a reaction center, where the first charge separation process takes place. Here we show experimental evidences that green algae (Chlorella vulgaris) in vivo display a synergistic two-photon absorption enhancement in their photosynthetic light harvesting. Their absorption coefficients at various wavelengths display dramatic dependence on the photon flux. This newly found phenomenon is attributed to a coherence-electronic-energy-transfer-mediated (CEETRAM) photon absorption process of light-harvesting pigment-protein complexes of green algae. Under the ambient light level, algae and higher plants can utilize this quantum mechanical mechanism to create two entangled electronic excitations adjacently in their light-harvesting networks. Concerted multiple electron transfer reactions in the reaction centers and oxygen evolving complexes can be implemented efficiently by the coherent motion of two entangled excitons from antennae to the charge separation reaction sites. To fabricate nanostructured, synthetic light-harvesting apparatus, the paramount role of the CEETRAM photon absorption mechanism should be seriously considered in the strategic guidelines.

  15. Mitochondrial Dynamics Tracking with Two-Photon Phosphorescent Terpyridyl Iridium(III) Complexes

    Science.gov (United States)

    Huang, Huaiyi; Zhang, Pingyu; Qiu, Kangqiang; Huang, Juanjuan; Chen, Yu; Ji, Liangnian; Chao, Hui

    2016-02-01

    Mitochondrial dynamics, including fission and fusion, control the morphology and function of mitochondria, and disruption of mitochondrial dynamics leads to Parkinson’s disease, Alzheimer’s disease, metabolic diseases, and cancers. Currently, many types of commercial mitochondria probes are available, but high excitation energy and low photo-stability render them unsuitable for tracking mitochondrial dynamics in living cells. Therefore, mitochondrial targeting agents that exhibit superior anti-photo-bleaching ability, deep tissue penetration and intrinsically high three-dimensional resolutions are urgently needed. Two-photon-excited compounds that use low-energy near-infrared excitation lasers have emerged as non-invasive tools for cell imaging. In this work, terpyridyl cyclometalated Ir(III) complexes (Ir1-Ir3) are demonstrated as one- and two-photon phosphorescent probes for real-time imaging and tracking of mitochondrial morphology changes in living cells.

  16. Efficient two-photon sensitized luminescence of europium (Ⅲ) complex based on hypersensitive transitions

    Institute of Scientific and Technical Information of China (English)

    Meng Shi; Hua Li; Mei Pan; Fufang Su; Lili Ma; Peigao Han; Hezhou Wang

    2011-01-01

    Red frequency-upconversion fluorescence emission is observed in europium(Ⅲ) complex with encapsulating polybenzimidazole tripodal ligands, pumped with 930- and 1070-nm picosecond laser pulses. The luminescence of transition 5D0 →7F2 (612 nm) is induced by two-photon absorption of hypersensitive transitions 7F0 →5D2 (465 nm) and 7F1 →5D1 (535 nm). Analysis results suggest that the two-photon excitation strength of these hypersensitive transitions is increased dramatically owing to the C3 symmetry of the coordination field.%@@ Red frequency-upconversion fluorescence emission is observed in europium(Ⅲ) complex with encapsulating polybenzimidazole tripodal ligands, pumped with 930- and 1070-nm picosecond laser pulses.The luminescence of transition 5D0 →7F2 (612 nm) is induced by two-photon absorption of hypersensitive transitions 7F0 →5D2 (465 nm) and 7F1 →5D1 (535 nm).Analysis results suggest that the two-photon excitation strength of these hypersensitive transitions is increased dramatically owing to the Ca symmetry of the coordination field.

  17. Steady state and time-resolved autofluorescence studies of human colonic tissues

    Institute of Scientific and Technical Information of China (English)

    Buhong Li; Zhenxi Zhang; Shusen Xie

    2006-01-01

    Steady state and time-resolved autofluorescence spectroscopies are employed to study the autofluorescence characteristics of human colonic tissues in vitro. The excitation wavelength varies from 260 to 540 nm, and the corresponding fluorescence emission spectra are acquired from 280 to 800 nm. Significant difference in fluorescence intensity of excitation-emission matrices (EEMs) is observed between normal and tumor colonic tissues. Compared with normal colonic tissue, low nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and flavin adenine dinucleotide (FAD), and high amino acids and protoporphyrin Ⅸ (PpⅨ) fluorescences characterize high-grade malignant tissue. Moreover, the autofluorescence lifetimes of normal and carcinomatous colonic tissues at 635 nm under 397-nm excitation are about 4.32±0.12 and 18.45±0.05 ns, respectively. The high accumulation of endogenous PpⅨ in colonic cancers is demonstrated in both steady state and time-resolved autofluorescence spectroscopies.

  18. Several Organic Salts with High Two-Photon Active

    Institute of Scientific and Technical Information of China (English)

    TIAN, Yu-Peng; JIANG, Min-Hua; WANG, He-Zhou; FANG, Qi

    2001-01-01

    Several organic salts with D-A molecular structure and different counterion have been prepared and experimentally investigated. The two-photon induced frequency-upconverted spectra and two-photon pumped lasing are measured for the organic salt solutions in various solvents. The results indicate that counterions have influence on their stability and lasing property.

  19. Two-photon holographic optogenetics of neural circuits (Conference Presentation)

    Science.gov (United States)

    Yang, Weijian; Carrillo-Reid, Luis; Peterka, Darcy S.; Yuste, Rafael

    2016-03-01

    Optical manipulation of in vivo neural circuits with cellular resolution could be important for understanding cortical function. Despite recent progress, simultaneous optogenetic activation with cellular precision has either been limited to 2D planes, or a very small numbers of neurons over a limited volume. Here we demonstrate a novel paradigm for simultaneous 3D activation using a low repetition rate pulse-amplified fiber laser system and a spatial light modulator (SLM) to project 3D holographic excitation patterns on the cortex of mice in vivo for targeted volumetric 3D photoactivation. This method is compatible with two-photon imaging, and enables the simultaneous activation of multiple cells in 3D, using red-shifted opsins, such as C1V1 or ReaChR, while simultaneously imaging GFP-based sensors such as GCaMP6. This all-optical imaging and 3D manipulation approach achieves simultaneous reading and writing of cortical activity, and should be a powerful tool for the study of neuronal circuits.

  20. Two-photon spectroscopy of trapped HD$^+$ ions in the Lamb-Dicke regime

    CERN Document Server

    Tran, Vu Quang; Douillet, Albane; Koelemeij, Jeroen C J; Hilico, Laurent

    2013-01-01

    We study the feasibility of nearly-degenerate two-photon rovibrational spectroscopy in ensembles of trapped, sympathetically cooled hydrogen molecular ions using a resonance-enhanced multiphoton dissociation (REMPD) scheme. Taking advantage of quasi-coincidences in the rovibrational spectrum, the excitation lasers are tuned close to an intermediate level to resonantly enhance two-photon absorption. Realistic simulations of the REMPD signal are obtained using a four-level model that takes into account saturation effects, ion trajectories, laser frequency noise and redistribution of population by blackbody radiation. We show that the use of counterpropagating laser beams enables optical excitation in an effective Lamb-Dicke regime. Sub-Doppler lines having widths in the 100 Hz range can be observed with good signal-to-noise ratio for an optimal choice of laser detunings. Our results indicate the feasibility of molecular spectroscopy at the $10^{-14}$ accuracy level for improved tests of molecular QED, a new det...

  1. Terahertz-visible two-photon rotational spectroscopy of cold OD-

    CERN Document Server

    Lee, Seunghyun; Lakhmanskaya, Olga; Spieler, Steffen; Endres, Eric S; Geistlinger, Katharina; Kumar, Sunil S; Wester, Roland

    2016-01-01

    We present a method to measure rotational transitions of molecular anions in the terahertz domain by sequential two-photon absorption. Ion excitation by bound-bound terahertz absorption is probed by absorption in the visible on a bound-free transition. The visible frequency is tuned to a state-selective photodetachment transition of the excited anions. This provides a terahertz action spectrum for just few hundred molecular ions. To demonstrate this we measure the two lowest rotational transitions, J=1<-0 and J =2<-1 of OD- anions in a cryogenic 22-pole trap. We obtain rotational transition frequencies of 598596.08(19) MHz for J=1<-0 and 1196791.57(27) MHz for J=2<-1 of OD-, in good agreement with their only previous measurement. This two-photon scheme opens up terahertz rovibrational spectroscopy for a range of molecular anions, in particular for polyatomic and cluster anions.

  2. Enhancement of Two-photon Absorption by Ce3+ Sensitization in Organic Dyes

    Institute of Scientific and Technical Information of China (English)

    LI Jian-fu; SUN Cheng-lin; ZHOU Hai-ling; XU Li-hua; YANG Qing-xin; JIANG Zhan-kui

    2007-01-01

    The two-photon absorption (TPA) and TPA-induced frequency upconversion emission properties of the dyes4-[P-(dicyanoethylamino) crystal]-N-methypyrdinium iodide and the complex of 4-[ P-(dicyanoethylamino) crystal]-N-methypyrdinium iodide and Ce( NO3 )3 were experimentally studied. It was found that the TPA cross section for the dye sensitized by Ce3+ is two factors larger than that of the dye without being sensitized. A three-level system model of the dye molecules was used to analyze the enhancement of TPA by the sensitizer Ce3+, which indicated that the sensitizer results in the increase of the transition dipole moment from the one-photon allowed excited state(1Bu)to the two-photon allowed excited state(2Ag).

  3. Dynamical modeling of pulsed two-photon interference

    Science.gov (United States)

    Fischer, Kevin A.; Müller, Kai; Lagoudakis, Konstantinos G.; Vučković, Jelena

    2016-11-01

    Single-photon sources are at the heart of quantum-optical networks, with their uniquely quantum emission and phenomenon of two-photon interference allowing for the generation and transfer of nonclassical states. Although a few analytical methods have been briefly investigated for describing pulsed single-photon sources, these methods apply only to either perfectly ideal or at least extremely idealized sources. Here, we present the first complete picture of pulsed single-photon sources by elaborating how to numerically and fully characterize non-ideal single-photon sources operating in a pulsed regime. In order to achieve this result, we make the connection between quantum Monte-Carlo simulations, experimental characterizations, and an extended form of the quantum regression theorem. We elaborate on how an ideal pulsed single-photon source is connected to its photocount distribution and its measured degree of second- and first-order optical coherence. By doing so, we provide a description of the relationship between instantaneous source correlations and the typical experimental interferometers (Hanbury-Brown and Twiss, Hong-Ou-Mandel, and Mach-Zehnder) used to characterize such sources. Then, we use these techniques to explore several prototypical quantum systems and their non-ideal behaviors. As an example numerical result, we show that for the most popular single-photon source—a resonantly excited two-level system—its error probability is directly related to its excitation pulse length. We believe that the intuition gained from these representative systems and characters can be used to interpret future results with more complicated source Hamiltonians and behaviors. Finally, we have thoroughly documented our simulation methods with contributions to the Quantum Optics Toolbox in Python in order to make our work easily accessible to other scientists and engineers.

  4. Synthesis of Two-Photon Materials and Two-Photon Liquid Crystals

    Science.gov (United States)

    Subramaniam, Girija

    2001-01-01

    The duration of the grant was interrupted by two major accidents that the PI met with-- an auto accident in Pasadena, CA during her second summer at JPL which took almost eight months for recovery and a second accident during Fall 2000 that left her in crutches for the entire semester. Further, the time released agreed by the University was not given in a timely fashion. The candidate has been given post-grant expire time off. In spite of all these problems, the PI synthesized a number of new two-photon materials and studied the structure-activity correlation to arrive at the best-optimized structure. The PI's design proved to be one of the best in the sense that these materials has a hitherto unreported two-photon absorption cross section. Many materials based on PI's design was later made by the NASA colleague. This is Phase 1. Phase II of this grant is to orate liquid crystalline nature into this potentially useful materials and is currently in progress. Recent observations of nano- and pico-second response time of homeotropically aligned liquid crystals suggest their inherent potentials to act as laser hardening materials, i.e., as protective devices against short laser pulses. The objective of the current project is to exploit this potential by the synthesis of liquid crystals with high optical nonlinearity and optimizing their performance. The PI is trying structural variations to bring in liquid crystalline nature without losing the high two-photon cross section. Both Phase I and Phase II led to many invited presentations and publications in reputed journals like 'Science' and 'Molecular Crystals'. The list of presentations and reprints are enclosed. Another important and satisfying outcome of this grant is the opportunity that this grant offered to the budding undergraduate scientists to get involved in a visible research of international importance. All the students had a chance to learn a lot during research, had the opportunity to present their work at

  5. Extreme nonlinearities in InAs/InP nanowire gain media: the two-photon induced laser

    DEFF Research Database (Denmark)

    Capua, Amir; Kami, Ouri; Eisenstein, Gadi;

    2012-01-01

    We demonstrate a novel laser oscillation scheme in an InAs / InP wire-like quantum dash gain medium. A short optical pulse excites carriers by two photon absorption which relax to the energy levels providing gain thereby enabling laser oscillations. The nonlinear dynamic interaction is analyzed...... and quantified using multi-color pump-probe measurements and shows a highly efficient nonlinear two photon excitation process which is larger by more than an order of magnitude compared to common quantum well and bulk gain media. The dynamic response of the nonlinearly induced laser line is characterized...

  6. Autofluorescence of pigmented skin lesions using a pulsed UV laser with synchronized detection: clinical results

    DEFF Research Database (Denmark)

    Cheng, Haynes Pak Hay; Svenmarker, Pontus; Tidemand-Lichtenberg, Peter;

    2010-01-01

    We report preliminary clinical results of autofluorescence imaging of malignant and benign skin lesions, using pulsed 355 nm laser excitation with synchronized detection. The novel synchronized detection system allows high signal-to-noise ratio to be achieved in the resulting autofluorescence...... signal, which may in turn produce high contrast images that improve diagnosis, even in the presence of ambient room light. The synchronized set-up utilizes a compact, diode pumped, pulsed UV laser at 355 nm which is coupled to a CCD camera and a liquid crystal tunable filter. The excitation and image...... capture is sampled at 5 kHz and the resulting autofluorescence is captured with the liquid crystal filter cycling through seven wavelengths between 420 nm and 580 nm. The clinical study targets pigmented skin lesions and evaluates the prospects of using autofluorescence as a possible means...

  7. Two-photon processes in highly charged ions

    Energy Technology Data Exchange (ETDEWEB)

    Jahrsetz, Thorsten

    2015-03-05

    Two-photon processes are atomic processes in which an atom interacts simultaneously with two photons. Such processes describe a wide range of phenomena, such as two-photon decay and elastic or inelastic scattering of photons. In recent years two-photon processes involving highly charged heavy ions have become an active area of research. Such studies do not only consider the total transition or scattering rates but also their angular and polarization dependence. To support such examinations in this thesis I present a theoretical framework to describe these properties in all two-photon processes with bound initial and final states and involving heavy H-like or He-like ions. I demonstrate how this framework can be used in some detailed studies of different two-photon processes. Specifically a detailed analysis of two-photon decay of H-like and He-like ions in strong external electromagnetic fields shows the importance of considering the effect of such fields for the physics of such systems. Furthermore I studied the elastic Rayleigh as well as inelastic Raman scattering by heavy H-like ions. I found a number of previously unobserved phenomena in the angular and polarization dependence of the scattering cross-sections that do not only allow to study interesting details of the electronic structure of the ion but might also be useful for the measurement of weak physical effects in such systems.

  8. Two-photon interference of temporally separated photons

    Science.gov (United States)

    Kim, Heonoh; Lee, Sang Min; Moon, Han Seb

    2016-10-01

    We present experimental demonstrations of two-photon interference involving temporally separated photons within two types of interferometers: a Mach-Zehnder interferometer and a polarization-based Michelson interferometer. The two-photon states are probabilistically prepared in a symmetrically superposed state within the two interferometer arms by introducing a large time delay between two input photons; this state is composed of two temporally separated photons, which are in two different or the same spatial modes. We then observe two-photon interference fringes involving both the Hong-Ou-Mandel interference effect and the interference of path-entangled two-photon states simultaneously in a single interferometric setup. The observed two-photon interference fringes provide simultaneous observation of the interferometric properties of the single-photon and two-photon wavepackets. The observations can also facilitate a more comprehensive understanding of the origins of the interference phenomena arising from spatially bunched/anti-bunched two-photon states comprised of two temporally separated photons within the interferometer arms.

  9. Quantum homodyne tomography of a two-photon Fock state

    CERN Document Server

    Ourjoumtsev, A; Grangier, P; Ourjoumtsev, Alexei; Tualle-Brouri, Rosa; Grangier, Philippe

    2006-01-01

    We present a continuous-variable experimental analysis of a two-photon Fock state of free-propagating light. This state is obtained from a pulsed non-degenerate parametric amplifier, which produces two intensity-correlated twin beams. Counting two photons in one beam projects the other beam in the desired two-photon Fock state, which is analyzed by using a pulsed homodyne detection. The Wigner function of the measured state is clearly negative. We developed a detailed analytic model which allows a fast and efficient analysis of the experimental results.

  10. Quantum homodyne tomography of a two-photon Fock state.

    Science.gov (United States)

    Ourjoumtsev, Alexei; Tualle-Brouri, Rosa; Grangier, Philippe

    2006-06-02

    We present a continuous-variable experimental analysis of a two-photon Fock state of free-propagating light. This state is obtained from a pulsed nondegenerate parametric amplifier, which produces two intensity-correlated twin beams. Counting two photons in one beam projects the other beam in the desired two-photon Fock state, which is analyzed by using a pulsed homodyne detection. The Wigner function of the measured state is clearly negative. We developed a detailed analytic model which allows a fast and efficient analysis of the experimental results.

  11. Scattering of two photons from two distant qubits: exact solution

    Energy Technology Data Exchange (ETDEWEB)

    Laakso, Matti; Pletyukhov, Mikhail [Institute for Theory of Statistical Physics, RWTH Aachen, 52056 Aachen (Germany)

    2015-07-01

    We consider the inelastic scattering of two photons from two qubits separated by an arbitrary distance and coupled to a one-dimensional transmission line. We present an exact, analytical solution to the problem, and use it to explore a particular configuration of qubits which is transparent to single-photon scattering, thus highlighting non-Markovian effects of inelastic two-photon scattering: Strong two-photon interference and momentum dependent photon (anti)bunching. This latter effect can be seen as an inelastic generalization of the Hong-Ou-Mandel effect.

  12. Intravital two-photon imaging: a versatile tool for dissecting the immune system.

    Science.gov (United States)

    Ishii, Taeko; Ishii, Masaru

    2011-03-01

    During the past decade, multi-photon or 'two-photon' excitation microscopy has launched a new era in the field of biological imaging. The near-infrared excitation laser for two-photon microscopy can penetrate thicker specimens, enabling the visualisation of living cell behaviour deep within tissues and organs without thin sectioning. The minimised photobleaching and toxicity enables the visualisation of live and intact specimens for extended periods. In this brief review, recent findings in intravital two-photon imaging for the physiology and pathology of the immune system are discussed. The immune system configures highly dynamic networks, where many cell types actively travel throughout the body and interact with each other in specific areas. Hence, real-time intravital imaging may be a powerful tool for dissecting the mechanisms of this dynamic system. The most unique characteristic of the immune system is its highly dynamic nature. A variety of cell types, such as lymphocytes, macrophages and dendritic cells (DCs), are continuously circulating throughout the body, migrating through the peripheral tissues and interacting with each other in their respective niches. Conventional methodologies in immunology, such as flow cytometry, cell or tissue culture, biochemistry and histology, have brought tremendous achievement within this field, although the dynamics of immune cells in an entire animal remain less clear. Technological progress of fluorescence microscopy has enabled us to visualise the intact biological phenomenon that has been uninvestigated. Among the advancements, the recent emergence and prevalence of two-photon, excitation-based, laser microscopy has revolutionised the research field, such that the dynamic behaviour of cells deep inside living organs can be visualised and analysed.

  13. RuBi-Glutamate: Two-photon and visible-light photoactivation of neurons and dendritic spines

    Directory of Open Access Journals (Sweden)

    Elodie Fino

    2009-05-01

    Full Text Available We describe neurobiological applications of RuBi-Glutamate, a novel caged-glutamate compound based on ruthenium photochemistry. RuBi-Glutamate can be excited with visible wavelengths and releases glutamate after one- or two-photon excitation. It has high quantum efficiency and can be used at low concentrations, partly avoiding the blockade of GABAergic transmission present with other caged compounds. Two-photon uncaging of RuBi-glutamate has a high spatial resolution and generates excitatory responses in individual dendritic spines with physiological kinetics. With laser beam multiplexing, RuBi-Glutamate uncaging can also be used to depolarize and fire pyramidal neurons with single-cell resolution. RuBi-Glutamate therefore enables the photo-activation of neuronal dendrites and circuits with visible or two-photon light sources, achieving single spine, or single cell, precision.

  14. Steady state anisotropy two-photon microscopy resolves multiple, spectrally similar fluorophores, enabling in vivo multilabel imaging.

    Science.gov (United States)

    Dubach, J Matthew; Vinegoni, Claudio; Weissleder, Ralph

    2014-08-01

    The use of spectrally distinguishable fluorescent dyes enables imaging of multiple targets. However, in two-photon microscopy, the number of fluorescent labels with distinct emission spectra that can be effectively excited and resolved is constrained by the confined tuning range of the excitation laser and the broad and overlapping nature of fluorophore two-photon absorption spectra. This limitation effectively reduces the number of available imaging channels. Here, we demonstrate that two-photon steady state anisotropy imaging (2PSSA) offers the capability to resolve otherwise unresolvable fluorescent tracers both in live cells and in mouse tumor models. This approach expands the number of biological targets that can be imaged simultaneously, increasing the total amount of information that can be obtained through imaging.

  15. NLO Electroweak Corrections to Higgs Decay to Two Photons

    OpenAIRE

    Actis, Stefano

    2009-01-01

    The recent calculation of the next-to-leading order electroweak corrections to the decay of the Standard Model Higgs boson to two photons in the framework of the complex-mass scheme is briefly summarized.

  16. Standard Model Higgs decay for two Photons in CMS

    CERN Multimedia

    Daniel Denegri

    2000-01-01

    Simulated two-photon mass distribution for SM Higgs and expected background in the CMS PbW04 crystal calorimeter for an integrated luminosity of 10 . 5 pb-1, with detailed simulation of calorimeter response.

  17. Synthesis of a Series of Novel Organic Compounds with Two-photon Absorption and Two-photon pumped Lasing

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    A series of novel organic compounds named as CSPI, DPASPI, PSPI DEASPI and HEASPI respectively, with large two-photon absorption has been synthesized and their structures have been determined by 1HNMR and elemental analysis. The highest two-photon pumped (TPP) output /input efficiency is as high as 13.4% for PSPI in DMF with d0 = 0.03 mol/L and the effective two-photon absorption cross section is 8.8′10-48 cm4×s/photon for DPASPI in DMF with d0= 0.05mol/L.

  18. Mass distribution for the two-photon channel

    CERN Multimedia

    ATLAS, collaboration

    2012-01-01

    Mass distribution for the two-photon channel. The strongest evidence for this new particle comes from analysis of events containing two photons. The smooth dotted line traces the measured background from known processes. The solid line traces a statistical fit to the signal plus background. The new particle appears as the excess around 126.5 GeV. The full analysis concludes that the probability of such a peak is three chances in a million.

  19. Comparison of calcium imaging in dorsal root ganglion neurons by using laser scanning confocal and two-photon microscopy

    Science.gov (United States)

    Huang, Yimei; Yang, Hongqin; Chen, Jiangxu; Shen, Xiuqiu; Zheng, Liqin; Wang, Yuhua; Xie, Shusen

    2012-03-01

    As one of the most important second messengers, calcium in nerve cells plays a critical role in neuronal processes, including excitability, neurotransmitter release, synaptic plasticity. Modulation of the calcium concentration is an important means of regulating diverse neuronal functions. To evaluate the role of calcium, quantitative measurement of cytosolic free calcium concentrations is necessary. There are several optical techniques that are available for measurement of calcium in live cells. Laser scanning confocal microscopy and two-photon microscopy are two prevalent techniques for their advantage in spatial resolution. In this paper, calcium in dorsal root ganglion neurons was imaged by laser scanning confocal microscopy and two-photon microscopy with Fluo-3, a calcium specific fluorescence probe. Both of spatial resolution and photobleaching, two common limitations of optical image modality, were compared between laser scanning confocal microscopy and two-photon microscopy, respectively. Three dimension images showed that laser scanning confocal microscopy and two-photon microscopy had not only similar lateral resolution but also parallel vertical resolution. However, Laser scanning confocal microscopy had an advantage over the two-photon microcopy in photobleaching. These results indicated that laser scanning confocal microscopy was more suitable than two-photon microscopy to be applied in imaging calcium in dorsal root ganglion neurons with Fluo-3.

  20. Sensing for intracellular thiols by water-insoluble two-photon fluorescent probe incorporating nanogel

    Energy Technology Data Exchange (ETDEWEB)

    Guo, Xudong; Zhang, Xin; Wang, Shuangqing; Li, Shayu [Beijing National Laboratory for Molecular Sciences, Key laboratory of Photochemistry, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190 (China); Hu, Rui, E-mail: hurui@iccas.ac.cn [Beijing National Laboratory for Molecular Sciences, Key laboratory of Photochemistry, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190 (China); Li, Yi, E-mail: yili@mail.ipc.ac.cn [Key Laboratory of Photochemical Conversion and Optoelectronic Materials, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, Beijing 100190 (China); Yang, Guoqiang, E-mail: gqyang@iccas.ac.cn [Beijing National Laboratory for Molecular Sciences, Key laboratory of Photochemistry, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190 (China)

    2015-04-15

    Highlights: • A novel “turn-on” two-photon fluorescent probe based on a π-conjugated triarylboron luminogen was designed and synthesized. • Fast, selective and sensitive detection of biothiols in 100% aqueous solution by simply loaded on a nanogel. • Single-photon and two-photon fluorescent bioimaging of biothiols in NIH/3T3 fibroblasts. - Abstract: A novel “turn-on” two-photon fluorescent probe containing a π-conjugated triarylboron luminogen and a maleimide moiety DMDP-M based on the photo-induced electron transfer (PET) mechanism for biothiol detection was designed and synthesized. By simply loading the hydrophobic DMDP-M on a cross-linked Pluronic{sup ®} F127 nanogel (CL-F127), a probing system DMDP-M/CL-F127 was established, which shows quick response, high selectivity and sensitivity to cysteine (Cys), homocysteine (Hcy) and glutathione (GSH) in aqueous phase. The DMDP-M/CL-F127 system presented the fastest response to Cys with a rate constant of 0.56 min{sup −1}, and the detection limit to Cys was calculated to be as low as 0.18 μM. The DMDP-M/CL-F127 system has been successfully applied to the fluorescence imaging of biothiols in NIH/3T3 fibroblasts either with single-photon or two-photon excitation because of its high biocompatibility and cell-membrane permeability. The present work provides a general, simple and efficient strategy for the application of hydrophobic molecules to sensing biothiols in aqueous phase, and a novel sensing system for intracellular biothiols fitted for both single-photon and two-photon fluorescence imaging.

  1. Phonon-Assisted Two-Photon Interference from Remote Quantum Emitters.

    Science.gov (United States)

    Reindl, Marcus; Jöns, Klaus D; Huber, Daniel; Schimpf, Christian; Huo, Yongheng; Zwiller, Val; Rastelli, Armando; Trotta, Rinaldo

    2017-07-12

    Photonic quantum technologies are on the verge of finding applications in everyday life with quantum cryptography and quantum simulators on the horizon. Extensive research has been carried out to identify suitable quantum emitters and single epitaxial quantum dots have emerged as near-optimal sources of bright, on-demand, highly indistinguishable single photons and entangled photon-pairs. In order to build up quantum networks, it is essential to interface remote quantum emitters. However, this is still an outstanding challenge, as the quantum states of dissimilar "artificial atoms" have to be prepared on-demand with high fidelity and the generated photons have to be made indistinguishable in all possible degrees of freedom. Here, we overcome this major obstacle and show an unprecedented two-photon interference (visibility of 51 ± 5%) from remote strain-tunable GaAs quantum dots emitting on-demand photon-pairs. We achieve this result by exploiting for the first time the full potential of a novel phonon-assisted two-photon excitation scheme, which allows for the generation of highly indistinguishable (visibility of 71 ± 9%) entangled photon-pairs (fidelity of 90 ± 2%), enables push-button biexciton state preparation (fidelity of 80 ± 2%) and outperforms conventional resonant two-photon excitation schemes in terms of robustness against environmental decoherence. Our results mark an important milestone for the practical realization of quantum repeaters and complex multiphoton entanglement experiments involving dissimilar artificial atoms.

  2. Reference Values of Skin Autofluorescence

    NARCIS (Netherlands)

    Koetsier, M.; Lutgers, H. L.; de Jonge, C.; Links, T. P.; Smit, A. J.; Graaff, R.

    2010-01-01

    Background: Skin autofluorescence (AF) as measured with the AGE Reader (DiagnOptics Technologies, Groningen, The Netherlands) is a noninvasive prognostic marker in diabetes mellitus and other diseases with increased cardiovascular risk. This study provides reference values of healthy Caucasian contr

  3. In Vivo Monitoring of Multiple Circulating Cell Populations Using Two-photon Flow Cytometry.

    Science.gov (United States)

    Tkaczyk, Eric R; Zhong, Cheng Frank; Ye, Jing Yong; Myc, Andrzej; Thomas, Thommey; Cao, Zhengyi; Duran-Struuck, Raimon; Luker, Kathryn E; Luker, Gary D; Norris, Theodore B; Baker, James R

    2008-02-15

    To detect and quantify multiple distinct populations of cells circulating simultaneously in the blood of living animals, we developed a novel optical system for two-channel, two-photon flow cytometry in vivo. We used this system to investigate the circulation dynamics in live animals of breast cancer cells with low (MCF-7) and high (MDA-MB-435) metastatic potential, showing for the first time that two different populations of circulating cells can be quantified simultaneously in the vasculature of a single live mouse. We also non-invasively monitored a population of labeled, circulating red blood cells for more than two weeks, demonstrating that this technique can also quantify the dynamics of abundant cells in the vascular system for prolonged periods of time. These data are the first in vivo application of multichannel flow cytometry utilizing two-photon excitation, which will greatly enhance our capability to study circulating cells in cancer and other disease processes.

  4. Two-Photon or Higher-Order Absorbing Optical Materials for Generation of Reactive Species

    Science.gov (United States)

    Cumpston, Brian (Inventor); Lipson, Matthew (Inventor); Marder, Seth R. (Inventor); Perry, Joseph W. (Inventor)

    2013-01-01

    Disclosed are highly efficient multiphoton absorbing compounds and methods of their use. The compounds generally include a bridge of pi-conjugated bonds connecting electron donating groups or electron accepting groups. The bridge may be substituted with a variety of substituents as well. Solubility, lipophilicity, absorption maxima and other characteristics of the compounds may be tailored by changing the electron donating groups or electron accepting groups, the substituents attached to or the length of the pi-conjugated bridge. Numerous photophysical and photochemical methods are enabled by converting these compounds to electronically excited states upon simultaneous absorption of at least two photons of radiation. The compounds have large two-photon or higher-order absorptivities such that upon absorption, one or more Lewis acidic species, Lewis basic species, radical species or ionic species are formed.

  5. Slow reflection and two-photon generation of microcavity exciton-polaritons

    CERN Document Server

    Steger, Mark; Snoke, David W; Pfeiffer, Loren; West, Ken

    2014-01-01

    We resonantly inject polaritons into a microcavity and track them in time and space as they feel a force due to the cavity gradient. This is an example of "slow reflection," as the polaritons, which can be viewed as renormalized photons, slow down to zero velocity and then move back in the opposite direction. These measurements accurately measure the lifetime of the polaritons in our samples, which is 180 $\\pm$ 10 ps, corresponding to a cavity leakage time of 135 ps and a cavity $Q$ of 320,000. Such long-lived polaritons propagate millimeters in these wedge-shaped microcavities. Additionally, we generate polaritons by two-photon excitation directly into the polariton states, allowing the possibility of modulation of the two-photon absorption by a polariton condensate.

  6. Dependence of the two-photon photoluminescence yield of gold nanostructures on the laser pulse duration

    Science.gov (United States)

    Biagioni, P.; Celebrano, M.; Savoini, M.; Grancini, G.; Brida, D.; Mátéfi-Tempfli, S.; Mátéfi-Tempfli, M.; Duò, L.; Hecht, B.; Cerullo, G.; Finazzi, M.

    2009-07-01

    Two-photon photoluminescence (TPPL) from gold nanostructures is becoming one of the most relevant tools for plasmon-assisted biological imaging and photothermal therapy as well as for the investigation of plasmonic devices. Here we study the yield of TPPL as a function of the temporal width δ of the excitation laser pulses for a fixed average power. In the δ>1ps regime, the TPPL yield decreases as δ is increased, while for shorter pulse widths it becomes independent of δ and, consequently, of the laser-pulse peak power. This peculiar dynamics is understood and modeled by considering that two-photon absorption in Au is a two-step process governed by the lifetime of the metastable state populated by the first photon absorption.

  7. Wide-field two-photon microscopy: features and advantages for biomedical applications

    Science.gov (United States)

    Wachsmann-Hogiu, S.; Hwang, J. Y.; Lindsley, E.; Farkas, D. L.

    2007-02-01

    We describe a simple fluorescence microscope based on wide-field two-photon excitation. While still taking advantage of some inherent properties of non-linear (two-photon) microscopy, such as increased penetration depth through tissue and reduced phototoxicity, this approach provides video frame rate imaging, can be easily coupled to fluorescence spectral and lifetime detection modules, and makes efficient use of the high average power currently available from ultrashort pulsed lasers. For a standard histopathology specimen, we were able to identify different structures based on spectral and fluorescence lifetime detection and analysis. We examined the use of 200fs and 2ps pulses from Spectra Physics MaiTai and Tsunami lasers, respectively, with average power ranging from 50mW to 500mW.

  8. Additive controlled synthesis of gold nanorods (GNRs) for two-photon luminescence imaging of cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Zhu Jing; Roy, Indrajit; Hu Rui; Ding Hong; Zhao Lingling; He, Guang S; Prasad, Paras N [Institute for Lasers, Photonics and Biophotonics, University at Buffalo, State University of New York, Buffalo, NY 14260-4200 (United States); Yong, Ken-Tye [School of Electrical and Electronic Engineering, Nanyang Technological University, Singapore 639798 (Singapore); Swihart, Mark T [Department of Chemical and Biological Engineering, University at Buffalo, State University of New York, Buffalo, NY 14260-4200 (United States); Cui Yiping, E-mail: ktyong@ntu.edu.sg, E-mail: cyp@seu.edu.cn, E-mail: pnprasad@buffalo.edu [Advanced Photonics Center, School of Electronic Science and Engineering, Southeast University, Nanjing 210096 (China)

    2010-07-16

    Gold nanorods (GNRs) with a longitudinal surface plasmon resonance peak that is tunable from 600 to 1100 nm have been fabricated in a cetyl trimethylammoniumbromide (CTAB) micellar medium using hydrochloric acid and silver nitrate as additives to control their shape and size. By manipulating the concentrations of silver nitrate and hydrochloric acid, the aspect ratio of the GNRs was reliably and reproducibly tuned from 2.5 to 8. The GNRs were first coated with polyelectrolyte multilayers and then bioconjugated to transferrin (Tf) to target pancreatic cancer cells. Two-photon imaging excited from the bioconjugated GNRs demonstrated receptor-mediated uptake of the bioconjugates into Panc-1 cells, overexpressing the transferrin receptor (TfR). The bioconjugated GNR formulation exhibited very low toxicity, suggesting that it is biocompatible and potentially suitable for targeted two-photon bioimaging.

  9. Additive controlled synthesis of gold nanorods (GNRs) for two-photon luminescence imaging of cancer cells.

    Science.gov (United States)

    Zhu, Jing; Yong, Ken-Tye; Roy, Indrajit; Hu, Rui; Ding, Hong; Zhao, Lingling; Swihart, Mark T; He, Guang S; Cui, Yiping; Prasad, Paras N

    2010-07-16

    Gold nanorods (GNRs) with a longitudinal surface plasmon resonance peak that is tunable from 600 to 1100 nm have been fabricated in a cetyl trimethylammoniumbromide (CTAB) micellar medium using hydrochloric acid and silver nitrate as additives to control their shape and size. By manipulating the concentrations of silver nitrate and hydrochloric acid, the aspect ratio of the GNRs was reliably and reproducibly tuned from 2.5 to 8. The GNRs were first coated with polyelectrolyte multilayers and then bioconjugated to transferrin (Tf) to target pancreatic cancer cells. Two-photon imaging excited from the bioconjugated GNRs demonstrated receptor-mediated uptake of the bioconjugates into Panc-1 cells, overexpressing the transferrin receptor (TfR). The bioconjugated GNR formulation exhibited very low toxicity, suggesting that it is biocompatible and potentially suitable for targeted two-photon bioimaging.

  10. Ag@Aggregation-induced emission dye core/shell nanostructures with enhanced one- and two-photon fluorescence

    Science.gov (United States)

    Wang, Cheng; Li, Yang; Xu, Qiujin; Luo, Liang

    2017-10-01

    Combining plasmonic nanostructures with two-photon fluorescence materials is a promising way to significantly enhance two-photon fluorescence. Ag@1,4-bis(2-cyano-2-phenylethenyl) benzene (BCPEB) core/shell nanostructures were fabricated by simply incubating the isolated Ag nanoparticles with BCPEB microrods in ethanol. BCPEB was chosen as the fluorescent organic molecule owing to the aggregation-induced-emission (AIE) nature which would reduce the emission loss as being practically applied in solid phase. By utilizing the match of the extinction spectrum of Ag nanoparticles and BCPEB's absorption band, the target Ag@BCPEB core/shell nanostructures showed an enhanced one-photon (12×) fluorescence, integrating with SERS signal as well. Moreover, the resultant second harmonic generation of Ag nanoparticles under two-photon excitation also well matched with the absorption band of BCPEB, and significant enhanced two-photon (17×) fluorescence was obtained. The confocal images of NIH-3T3 cells with these nanostructures under one- and two-photon excitation showed good contrast and brightness for bio-imaging.

  11. Two-photon spectroscopic behaviors and photodynamic effect on the BEL-7402 cancer cells of the new chlorophyll photosensitizer

    Institute of Scientific and Technical Information of China (English)

    ZHAO PeiDe; ZHANG GuiLan; CHEN WenJu; CHEN Ping; TANG GuoQing; LIU JinWei; LIN Lie; GUO Peng; YU Qing; YAO JianZhong; MA DongMing

    2008-01-01

    The spectroscopic properties of a new chlorophyll derivate photosensitizer (CDP) are studied under the excitation wavelengths at 800 and 400 nm using femtosecond pulses from a Ti: sapphire laser. The damaging effect of CDP on the BEL-7402 cancer cells is also investigated upon two-photon illumination at 800 nm. The normalized fluorescence spectra of CDP in tetrahydrofuran (THF) show that two-photon and one-photon spectra have the same distributions and the same emission bands (675 nm). The life-times of two- and one-photon induced fluorescence of this molecule are of the order of 5.0 ns. By comparing the data it is shown that there is some difference between the two lifetimes, but the differ-ence is less than one nanosecond. The two-photon absorption cross section of the molecule is also measured at 800 nm and estimated as about σ'2≈31.5×10-50 cm4·s·photon-1. The results of two-photon photodynamic therapy (TPPDT) tests show that CDP can kill all of the tested cancer cells according to the usual Eosine assessment. Our results indicate that the two-photon-induced photophysical, photochemical and photosensitizing processes of CDP may be basically similar to those of one-photon excitation. These behaviors of the sample suggest that one may find other possible methods to estimate some photosensitizers' effects in details such as their distribution in cells and the reactive targets of the sub-cellular parts of some tumor cells via two-photon excitation techniques.

  12. Fast, High Fidelity Quantum Dot Spin Initialization without a Strong Magnetic Field by Two-Photon Processes

    CERN Document Server

    Majumdar, Arka; Faraon, Andrei; Vuckovic, Jelena

    2009-01-01

    We describe a proposal for fast electron spin initialization in a negatively charged quantum dot coupled to a microcavity without the need for a strong magnetic field. We employ two-photon excitation to access trion states that are spin forbidden by one-photon excitation. Our simulation shows a maximum initialization speed of 1.3 GHz and maximum fidelity of 99.7% with realistic system parameters.

  13. Suppression and Enhancement in Parametric Two-Photon Resonant Nondegenerate Four-Wave Mixing via Quantum Interference

    Institute of Scientific and Technical Information of China (English)

    SUN Jiang; MI Xin; YU Zu-He; JIANG Qian; ZUO Zhan-Chun; WANG Yan-Bang; WU Ling-An; FU Pan-Ming

    2004-01-01

    @@ Quantum interference may lead to suppression and enhancement of the two-photon resonant nondegenerate fourwave mixing signal in a cascade four-level system. Such phenomena are demonstrated in Ba through inducing atomic coherence between the ground state 6s2 and the doubly excited autoionizing Rydberg state 6pnd. This method can be used as a new spectroscopic tool for measuring the transition dipole moment between two highly excited atomic states.

  14. Hyperspectral deep ultraviolet autofluorescence of muscle fibers is affected by postmortem changes.

    Science.gov (United States)

    Chagnot, Caroline; Vénien, Annie; Jamme, Frédéric; Réfrégiers, Matthieu; Desvaux, Mickaël; Astruc, Thierry

    2015-05-20

    After slaughter, muscle cells undergo biochemical and physicochemical changes that may affect their autofluorescence characteristics. The autofluorescent response of different rat extensor digitorum longus (EDL) and soleus muscle fiber types was investigated by deep ultraviolet (UV) synchrotron microspectroscopy immediately after animal sacrifice and after 24 h of storage in a moist chamber at 20 °C. The glycogen content decreased from 23 to 18 μmol/g of fresh muscle in 24 h postmortem. Following a 275 nm excitation wavelength, the spectral muscle fiber autofluorescence response showed discrimination depending upon postmortem time (t0 versus t24 h) on both muscles at 346 and 302 nm and, to a lesser extent, at 408 and 325 nm. Taken individually, all fiber types were discriminated but with variable accuracy, with type IIA showing better separation of t0/t24 h than other fiber types. These results suggest the usefulness of the autofluorescent response of muscle cells for rapid meat-aging characterization.

  15. Autofluorescence guided diagnostic evaluation of suspicious oral mucosal lesions: opportunities, limitations, and pitfalls

    Science.gov (United States)

    Vigneswaran, Nadarajah

    2011-03-01

    Wide-filed autofluorescence examination is currently considered as a standard of care for screening and diagnostic evaluation of early neoplastic changes of the skin, cervix, lung, bladder, gastrointestinal tract and oral cavity. Naturally occurring fluorophores within the tissue absorb UV and visible light and can re-emit some of this light at longer wavelengths in the form of fluorescence. This non-invasive tissue autofluorescence imaging is used in optical diagnostics, especially in the early detection of cancer. Usually, malignant transformation is associated with thickening of the epithelium, enhanced cellular density due to increased nuclear cytoplasmic ratio which may attenuate the excitation leading to a decrease in collagen autofluorescence. Hence, dysplastic and cancerous tissues often exhibit decreased blue-green autofluorescence and appear darker compared to uninvolved mucosa. Currently, there are three commercially available devices to examine tissue autofluorescence in the oral cavity. In this study we used the oral cancer screening device IdentafiTM 3000 to examine the tissue reflectance and autofluorescence of PML and confounding lesions of the oral cavity. Wide-field autofluorescence imaging enables rapid inspection of large mucosal surfaces, to aid in recognition of suspicious lesions and may also help in discriminate the PML (class 1) from some of the confounding lesions (class II). However, the presence of inflammation or pigments is also associated with loss of stromal autofluorescence, and may give rise to false-positive results with widefield fluorescence imaging. Clinicians who use these autofluorescence based oral cancer screening devices should be aware about the benign oral mucosal lesions that may give false positivity so that unnecessary patient's anxiety and the need for scalpel biopsy can be eliminated.

  16. Ultra-thin rigid endoscope: Two-photon imaging through a graded-index multi-mode fiber

    CERN Document Server

    Sivankutty, Siddharth; Cossart, Rosa; Bouwmans, Géraud; Monneret, Serge; Rigneault, Hervé

    2015-01-01

    Rigid endoscopes like graded-index (GRIN) lenses are known tools in biological imaging, but it is conceptually difficult to miniaturize them. In this letter, we demonstrate an ultra-thin rigid endoscope with a diameter of only 125 microns. In addition, we identify a domain where two-photon endoscopic imaging with fs-pulse excitation is possible. We validate the ultra-thin rigid endoscope consisting of a few cm of graded-index multi-mode fiber by using it to acquire optically sectioned two-photon fluorescence endoscopic images of three-dimensional samples.

  17. Nonlinear quantitative photoacoustic tomography with two-photon absorption

    CERN Document Server

    Ren, Kui

    2016-01-01

    Two-photon photoacoustic tomography (TP-PAT) is a non-invasive optical molecular imaging modality that aims at inferring two-photon absorption property of heterogeneous media from photoacoustic measurements. In this work, we analyze an inverse problem in quantitative TP-PAT where we intend to reconstruct optical coefficients in a semilinear elliptic PDE, the mathematical model for the propagation of near infra-red photons in tissue-like optical media with two-photon absorption, from the internal absorbed energy data. We derive uniqueness and stability results on the reconstructions of single and multiple optical coefficients, and present some numerical reconstruction results based on synthetic data to complement the theoretical analysis.

  18. Two-photon interference between disparate sources for quantum networking

    Science.gov (United States)

    McMillan, A. R.; Labonté, L.; Clark, A. S.; Bell, B.; Alibart, O.; Martin, A.; Wadsworth, W. J.; Tanzilli, S.; Rarity, J. G.

    2013-06-01

    Quantum networks involve entanglement sharing between multiple users. Ideally, any two users would be able to connect regardless of the type of photon source they employ, provided they fulfill the requirements for two-photon interference. From a theoretical perspective, photons coming from different origins can interfere with a perfect visibility, provided they are made indistinguishable in all degrees of freedom. Previous experimental demonstrations of such a scenario have been limited to photon wavelengths below 900 nm, unsuitable for long distance communication, and suffered from low interference visibility. We report two-photon interference using two disparate heralded single photon sources, which involve different nonlinear effects, operating in the telecom wavelength range. The measured visibility of the two-photon interference is 80 +/- 4%, which paves the way to hybrid universal quantum networks.

  19. Portable LED-induced autofluorescence spectroscopy for oral cancer diagnosis

    Science.gov (United States)

    Yan, Yung-Jhe; Huang, Ting-Wei; Cheng, Nai-Lun; Hsieh, Yao-Fang; Tsai, Ming-Hsui; Chiou, Jin-Chern; Duann, Jeng-Ren; Lin, Yung-Jiun; Yang, Chin-Siang; Ou-Yang, Mang

    2017-04-01

    Oral cancer is a serious and growing problem in many developing and developed countries. To improve the cancer screening procedure, we developed a portable light-emitting-diode (LED)-induced autofluorescence (LIAF) imager that contains two wavelength LED excitation light sources and multiple filters to capture ex vivo oral tissue autofluorescence images. Compared with conventional means of oral cancer diagnosis, the LIAF imager is a handier, faster, and more highly reliable solution. The compact design with a tiny probe allows clinicians to easily observe autofluorescence images of hidden areas located in concave deep oral cavities. The ex vivo trials conducted in Taiwan present the design and prototype of the portable LIAF imager used for analyzing 31 patients with 221 measurement points. Using the normalized factor of normal tissues under the excitation source with 365 nm of the central wavelength and without the bandpass filter, the results revealed that the sensitivity was larger than 84%, the specificity was not smaller than over 76%, the accuracy was about 80%, and the area under curve of the receiver operating characteristic (ROC) was achieved at about 87%, respectively. The fact shows the LIAF spectroscopy has the possibilities of ex vivo diagnosis and noninvasive examinations for oral cancer.

  20. Two-photon interference with non-identical photons

    Science.gov (United States)

    Liu, Jianbin; Zhou, Yu; Zheng, Huaibin; Chen, Hui; Li, Fu-li; Xu, Zhuo

    2015-11-01

    Two-photon interference with non-identical photons is studied based on the superposition principle in Feynman's path integral theory. The second-order temporal interference pattern is observed by superposing laser and pseudothermal light beams with different spectra. The reason why there is two-photon interference for photons of different spectra is that non-identical photons can be indistinguishable for the detection system when Heisenberg's uncertainty principle is taken into account. These studies are helpful to understand the second-order interference of light in the language of photons.

  1. Two-Photon Total Annihilation of Molecular Positronium

    CERN Document Server

    Pérez-Ríos, Jesús; Greene, Chris H

    2014-01-01

    The rate for complete two-photon annihilation of molecular positronium Ps$_{2}$ is reported. This decay channel involves a four-body collision among the fermions forming Ps$_{2}$, and two photons of 1.022 MeV, each, as the final state. The quantum electrodynamics result for the rate of this process is found to be $\\Gamma_{Ps_{2} \\rightarrow \\gamma\\gamma}$ = 9.0 $\\times 10^{-12}$ s$^{-1}$. This decay channel completes the most comprehensive decay chart for Ps$_{2}$ up to date.

  2. Two-photon Compton process in pulsed intense laser fields

    CERN Document Server

    Seipt, D

    2012-01-01

    Based on strong-field QED in the Furry picture we use the Dirac-Volkov propagator to derive a compact expression for the differential emission probability of the two-photon Compton process in a pulsed intense laser field. The relation of real and virtual intermediate states is discussed, and the natural regularization of the on-shell contributions due to the finite laser pulse is highlighted. The inclusive two-photon spectrum is two orders of magnitude stronger than expected from a perturbative estimate.

  3. Precision two-photon spectroscopy of alkali elements

    Indian Academy of Sciences (India)

    P V Kiran Kumar; M V Suryanarayana

    2014-08-01

    In this paper, we have briefly reviewed the work on two-photon spectroscopy of alkali elements and its applications. The technique of Doppler-free two-photon spectroscopy is briefly summarized. A review of various techniques adopted for measuring absolute frequencies of the atomic transitions and precision measurements of isotope shifts and hyperfine structures (HFS) is presented. Some of the recent works on precision measurements of HFS constants of 6 ${}^2S_{1/2}$ level of ${}^{39}$K and ${}^{41}$K, 9 ${}^2S_{1/2}$ level and 7 ${}^2D_{3/2}$ level of 133Cs are also discussed.

  4. A fluorescent benzothiazole probe with efficient two-photon absorption

    Science.gov (United States)

    Echevarria, Lorenzo; Moreno, Iván; Camacho, José; Salazar, Mary Carmen; Hernández, Antonio

    2012-11-01

    In this work, we report the two-photon absorption of 2-[4-(dimethylamino)phenyl]-1,3-benzothiazole-6-carbonitrile (DBC) in DMSO solution pumping at 779 nm with a 10 ns pulse laser-Nd:YAG system. The obtained two-photon absorption cross-section in DBC (407 ± 18 GM) is considerably high. Because DBC is a novel compound and have high values of fluorescence quantum yield, this result is expected to have an impact in biomolecules detection, diagnosis and treatment of cancer. Similar structures have previously been reported to show remarkable antitumour effects.

  5. Modulation of attosecond beating by resonant two-photon transition

    CERN Document Server

    Galán, Álvaro Jiménez; Martín, Fernando

    2015-01-01

    We present an analytical model that characterizes two-photon transitions in the presence of autoionising states. We applied this model to interpret resonant RABITT spectra, and show that, as a harmonic traverses a resonance, the phase of the sideband beating significantly varies with photon energy. This phase variation is generally very different from the $\\pi$ jump observed in previous works, in which the direct path contribution was negligible. We illustrate the possible phase profiles arising in resonant two-photon transitions with an intuitive geometrical representation.

  6. NIR autofluorescence and OCT imaging of biotissues

    Science.gov (United States)

    Chorvat, Dusan, Jr.; Smolka, Jozef; Mateasik, Anton; Hrin, Lubos

    2003-10-01

    Optical coherence tomography (OCT) is one of the most promising recently developed methods for non-invasive in vivo characterization of biological highly scattering tissues. However, one of the drawbacks of the pure OCT imaging is that it is not sensitive to changes in metabolism. This may impact derived information and consecutive diagnostics, because pathological changes of tissue structure are accompanied with changes in metabolic activity or functional state in these areas even if there is not yet strongly evident structural change. Therefore, it is desirable to combine early detection of tissue malformations by OCT with other techniques, capable to detect and evaluate their functional state. One of suitable candidates for such non-invasive optical functional imaging is detection of laser induced autofluorescence, which could provide information about rate of biological and chemical processes in living cells. As an example, the cells in proliferative state (with increased metabolic activity or during mitosis) show more intensive NIR fluorescence than the cells that are not proliferative, because of increased concentration of free porphyrins. In presented study we used OCT and laser induced NIR autofluorescence imaging for detection and evaluation of changes in areas of naevus and injuries in group of volunteers. The fusion of information on structural and functional state of biotissues provided by the two mentioned complementary methods may enhance the diagnostics power of their prospective clinical use. Firstly the fluorescence of area of naevus and injuries, excited by 630 nm was taken by CCD camera and then was naevus scanned by OCT. The findings of both methods were compared and correlated. In the case of naevus the obtained results were reviewed with histological treatment of the same area.

  7. Two-photon light-sheet nanoscopy by fluorescence fluctuation correlation analysis

    Science.gov (United States)

    Chen, Xuanze; Zong, Weijian; Li, Rongqin; Zeng, Zhiping; Zhao, Jia; Xi, Peng; Chen, Liangyi; Sun, Yujie

    2016-05-01

    Advances in light-sheet microscopy have enabled the fast three-dimensional (3D) imaging of live cells and bulk specimens with low photodamage and phototoxicity. Combining light-sheet illumination with super-resolution imaging is expected to resolve subcellular structures. Actually, such kind of super-resolution light-sheet microscopy was recently demonstrated using a single-molecule localization algorithm. However, the imaging depth and temporal resolution of this method are limited owing to the requirements of precise single molecule localization and reconstruction. In this work, we present two-photon super-resolution light-sheet imaging via stochastic optical fluctuation imaging (2PLS-SOFI), which acquires high spatiotemporal resolution and excellent optical sectioning ability. 2PLS-SOFI is based on non-linear excitation of fluctuation/blinking probes using our recently developed fast two-photon three-axis digital scanned light-sheet microscope (2P3A-DSLM), which enables both deep penetration and thin sheet of light. Overall, 2PLS-SOFI demonstrates up to 3-fold spatial resolution enhancement compared with conventional two-photon light-sheet (2PLS) microscopy and about 40-fold temporal resolution enhancement compared with individual molecule localization-selective plane illumination microscopy (IML-SPIM). Therefore, 2PLS-SOFI is promising for 3D long-term, deep-tissue imaging with high spatiotemporal resolution.

  8. Two-photon fluorescent sensor for K+ imaging in live cells (Conference Presentation)

    Science.gov (United States)

    Sui, Binglin; Yue, Xiling; Kim, Bosung; Belfield, Kevin D.

    2016-03-01

    It is difficult to overstate the physiological importance of potassium for life as its indispensable roles in a variety of biological processes are widely known. As a result, efficient methods for determining physiological levels of potassium are of paramount importance. Despite this, relatively few K+ fluorescence sensors have been reported, with only one being commercially available. A new two-photon excited fluorescent K+ sensor is reported. The sensor is comprised of three moieties, a highly selective K+ chelator as the K+ recognition unit, a boron-dipyrromethene (BODIPY) derivative modified with phenylethynyl groups as the fluorophore, and two polyethylene glycol chains to afford water solubility. The sensor displays very high selectivity (physiological metal cations. Upon binding K+, the sensor switches from non-fluorescent to highly fluorescent, emitting red to near-IR (NIR) fluorescence. The sensor exhibited a good two-photon absorption cross section, 500 GM at 940 nm. Moreover, it is not sensitive to pH in the physiological pH range. Time-dependent cell imaging studies via both one- and two-photon fluorescence microscopy demonstrate that the sensor is suitable for dynamic K+ sensing in living cells.

  9. Cell flow analysis with a two-photon fluorescence fiber probe

    Science.gov (United States)

    Chang, Yu-Chung; Ye, Jing Yong; Thomas, Thommey P.; Baker, James R., Jr.; Norris, Theodore B.

    2010-11-01

    We report the use of a sensitive double-clad fiber (DCF) probe for in situ cell flow velocity measurements and cell analysis by means of two-photon excited fluorescence correlation spectroscopy (FCS). We have demonstrated the feasibility to use this fiber probe for in vivo two-photon flow cytometry previously. However, because of the viscosity of blood and the non-uniform flow nature in vivo, it is problematic to use the detected cell numbers to estimate the sampled blood volume. To precisely calibrate the sampled blood volume, it is necessary to conduct real time flow velocity measurement. We propose to use FCS technique to measure the flow velocity. The ability to measure the flow velocities of labeled cells in whole blood has been demonstrated. Our two-photon fluorescence fiber probe has the ability to monitor multiple fluorescent biomarkers simultaneously. We demonstrate that we can distinguish differently labeled cells by their distinct features on the correlation curves. The ability to conduct in situ cell flow analysis using the fiber probe may be useful in disease diagnosis or further comprehension of the circulation system.

  10. Calculation of the spatial resolution in two-photon absorption spectroscopy applied to plasma diagnosis

    Energy Technology Data Exchange (ETDEWEB)

    Garcia-Lechuga, M. [Departamento de Física Teórica, Atómica y Óptica, Universidad de Valladolid, 47011-Valladolid (Spain); Laser Processing Group, Instituto de Óptica “Daza de Valdés,” CSIC, 28006-Madrid (Spain); Fuentes, L. M. [Departamento de Física Aplicada, Universidad de Valladolid, 47011-Valladolid (Spain); Grützmacher, K.; Pérez, C., E-mail: concha@opt.uva.es; Rosa, M. I. de la [Departamento de Física Teórica, Atómica y Óptica, Universidad de Valladolid, 47011-Valladolid (Spain)

    2014-10-07

    We report a detailed characterization of the spatial resolution provided by two-photon absorption spectroscopy suited for plasma diagnosis via the 1S-2S transition of atomic hydrogen for optogalvanic detection and laser induced fluorescence (LIF). A precise knowledge of the spatial resolution is crucial for a correct interpretation of measurements, if the plasma parameters to be analysed undergo strong spatial variations. The present study is based on a novel approach which provides a reliable and realistic determination of the spatial resolution. Measured irradiance distribution of laser beam waists in the overlap volume, provided by a high resolution UV camera, are employed to resolve coupled rate equations accounting for two-photon excitation, fluorescence decay and ionization. The resulting three-dimensional yield distributions reveal in detail the spatial resolution for optogalvanic and LIF detection and related saturation due to depletion. Two-photon absorption profiles broader than the Fourier transform-limited laser bandwidth are also incorporated in the calculations. The approach allows an accurate analysis of the spatial resolution present in recent and future measurements.

  11. Two-photon microscopy for imaging germinal centers and T follicular helper cells.

    Science.gov (United States)

    Clatworthy, Menna R

    2015-01-01

    One of the principle features of immune cells is their dynamic nature. Lymphocytes circulate in the blood between secondary lymphoid organs and tissues in an effort to maximize the likelihood of a rapid and appropriate immune response to invading pathogens and tissue damage. Conventional experimental techniques such as histology and flow cytometry have greatly increased our understanding of immune cells, but in the last decade, two-photon microscopy has revolutionized our ability to interrogate the dynamic behavior of immune cells, a facet so critical to their function. Two-photon microscopy relies on the excitation of fluorophores by simultaneous application of two photons of longer wavelength light. This allows a greater depth of imaging with minimal photodamage. Thus, living tissues can be imaged, including immune cells in lymph nodes. This technique has been used to interrogate the events occurring in a germinal center response and the interactions between cells in the germinal center, including T follicular helper cells (Tfh), germinal center B cells, and follicular dendritic cells (FDC). Herein, a method is described by which the interactions between Tfh and B cells within a germinal center in a popliteal lymph node can be imaged in a live mouse.

  12. Two-photon fluorescence and fluorescence imaging of two styryl heterocyclic dyes combined with DNA.

    Science.gov (United States)

    Gao, Chao; Liu, Shu-yao; Zhang, Xian; Liu, Ying-kai; Qiao, Cong-de; Liu, Zhao-e

    2016-03-01

    Two new styryl heterocyclic two-photon (TP) materials, 4-[4-(N-methyl)styrene]-imidazo [4,5-f][1,10] phenanthroline-benzene iodated salt (probe-1) and 4,4-[4-(N-methyl)styrene]-benzene iodated salt (probe-2) were successfully synthesized and studied as potential fluorescent probes of DNA detection. The linear and nonlinear photophysical properties of two compounds in different solvents were investigated. The absorption, one- and two-photon fluorescent spectra of the free dye and dye-DNA complex were also examined to evaluate their photophysical properties. The binding constants of dye-DNA were obtained according to Scatchard equation with good values. The results showed that two probes could be used as fluorescent DNA probes by two-photon excitation, and TP fluorescent properties of probe-1 are superior to that of probe-2. The fluorescent method date indicated that the mechanisms of dye-DNA complex interaction may be groove binding for probe-1 and electrostatic interaction for probe-2, respectively. The MTT assay experiments showed two probes are low toxicity. Moreover, the TP fluorescence imaging of DNA detection in living cells at 800 nm indicated that the ability to locate in cell nuclei of probe-1 is better than that of probe-2.

  13. Two-photon photodissociation of H 2O via the B˜ state

    Science.gov (United States)

    Underwood, J.; Wittig, C.

    2004-03-01

    The H 2O B˜← X˜ system has been excited by using 266 nm two-photon absorption, and H-atom products have been probed by using high- n Rydberg time-of-flight spectroscopy. The B˜/ X˜ conical intersection results in the efficient transfer of flux from B˜ to X˜, with dissociation occurring mainly on X˜ . Though the OH(X 2Π) product is highly rotationally excited, this bias is less than that observed at higher energies. The OH(A 2Σ +)/OH(X 2Π) branching ratio is <0.1%, which is significantly smaller than branching ratios obtained at higher excitation energies, and two orders of magnitude less than predicted by theory.

  14. Two-photon absorption-induced drug delivery from polymers for medical applications

    Science.gov (United States)

    Kim, Hee-Cheol; Kreiling, Stefan; Haertner, Sebastian; Hesse, Lutz; Greiner, Andreas; Hampp, Norbert A.

    2004-06-01

    Novel polymeric materials carrying a drug depot have been developed which are suitable for fabrication of photochemically modulated drug delivery devices. In order to avoid uncontrolled drug release the drug is covalently attached to the polymer backbone using a photo-active linker. Controlled drug release from the polymer can be accomplished either via single-photon excitation or by two-photon absorption (TPA). In particular the second possibility is of interest for applications where exposure to day light or UV light may not be omitted. One example are polymeric intraocular lenses (IOL), which are implanted instead of the opaque natural lens during cataract surgery. Secondary cataract formation is quite often observed after implantation of polymeric IOLs. In this study the well known cell toxic agent 5-fluorouracil (5FU) attached to a methylmethacrylate-based polymer was investigated as an IOL which can upon photochemical excitation release 5FU in order to treat or to prevent secondary cataract formation. The photochemical cleavage of the linker molecule was analyzed with single- and two-photon excitation. UV/VIS spectroscopy and HPLC analysis confirmed the release of 5FU form the polymer backbone. The diffusion of the drug precursor out from the polymer as well as the hydrolysis of the drug precursor which leads to 5FU formation were investigated in vitro.

  15. Atom-atom entanglement generated at early times by two-photon emission

    Energy Technology Data Exchange (ETDEWEB)

    Leon, Juan; Sabin, Carlos [Instituto de Fisica Fundamental, CSIC, Serrano 113-bis, 28006 Madrid (Spain)], E-mail: leon@imaff.cfmac.csic.es, E-mail: csl@imaff.cfmac.csic.es

    2009-07-15

    We analyze entanglement generation between a pair of neutral two-level atoms that are initially excited in a common electromagnetic vacuum. The nonlocal correlations that appear due to the interaction with the field can become entanglement when the field state is known. We distinguish two different situations: in the first, the field remains in the vacuum state and in the second, two photons are present in the final state. In both cases, we study the dependence of the entanglement on time and interatomic distance, at ranges related with locality issues.

  16. Laser fabrication of Au nanorod aggregates microstructures assisted by two-photon polymerization.

    Science.gov (United States)

    Masui, Kyoko; Shoji, Satoru; Asaba, Kenji; Rodgers, Thomas C; Jin, Feng; Duan, Xuan-Ming; Kawata, Satoshi

    2011-11-07

    We demonstrate fabrication of Au nanorod aggregates microstructures by means of a femtosecond near-infrared laser. The laser light was tightly focused into colloidal Au nanorods dispersed in photopolymerizable metyl-methacrylate (MMA) compound to induce two-photon polymerization (TPP). TPP of MMA glued the nanorods together to form solid microstrucures of aggregates. The laser light excited a local surface plasmon, resulting in confinement of TPP in the vicinity of nanorods. Concurrenly occurring optical accumulation of nanorods created a unique mechanism for the formation of nanorod aggregates into desired microstructures. This technique would be a clue for a novel micro/nanofabrication method for plasmonic materials and devices.

  17. Two-photon photodynamic properties of TBO-AuNR-in-shell nanoparticles (Conference Presentation)

    Science.gov (United States)

    Wu, Cheng-Han; Yeh, Chen-Sheng; Cheng, Fong-Yu; Tsai, Zen-Uong; Liu, Tzu-Ming

    2016-03-01

    Photodynamic therapy (PDT) is a light-activated chemotherapeutic treatment that utilizes singlet oxygen and reactive oxygen species induced oxidative reactions to react with surrounding biological substrates, which either kills or irreversibly damages malignant cells. We used multiphoton nonlinear optical microscopy to observe the photo-dynamic effects of TBO-AuNR-in-shell NPs. Excited by femtosecond Cr:forsterite laser operating at 1230nm, singlet oxygen were generated through a plasmon-enhanced two-photon nonlinear optical process. For cells took up NPs, this photodynamic effect can kill the cell. From nonlinear optical microscopy images, we found they shrunk after 3 minutes of illumination.

  18. Morphology dependent two photon absorption in plasmonic structures and plasmonic-organic hybrids

    Science.gov (United States)

    Gambhir, Kaweri; Ray, Bhumika; Mehrotra, Ranjana; Sharma, Parag

    2017-05-01

    Two photon absorption coefficients of two distinct plasmonic structures, namely, gold nanoflowers (GNF) and gold nanopebbles (GNP) have been investigated and compared with conventional gold nanospheres (GNS). All three different nanoshapes were synthesized by changing the reaction solvent under the same experimental procedure. Further, hybrids of these plasmonic structures were prepared with an organic dye Eosin yellow (EY), to investigate the morphology effect of plasmonic structures on plasmonic-organic hybrids in terms of their linear extinction spectra and two photon absorption coefficients. The NLO investigations were conducted using 20 ps laser pulses of wavelength 532 nm as an excitation source in single beam Z-scan setup. UV/visible spectroscopy was employed for monitoring plasmon resonances and changes in linear extinction spectra. The experimental outcomes revealed two photon absorption coefficients of EY increased 120%, 32% and 39%, while 69%, 60% and 53% enhancement in the peaks of linear extinction maxima of EY has been observed, when hybridized with GNF, GNS and GNP, respectively. This boost in the optical coefficients may be attributed to dimerization of EY molecules on the surface of nanoparticles. Keeping the toxicity of EY in view, we propose that the two photon absorption coefficients of this dye and control thereof, by the addition of plasmonic structures would be helpful not only in understanding the interactions between plasmons and fluorophore, but also pave an efficient way, to reduce the operative concentration of this hazardous dye in a wide range of applications and thereby, mitigating the environmental degradation caused by its highly concentrated effluents.

  19. Two-photon spectroscopic behaviors and photodynamic effect on the BEL-7402 cancer cells of the new chlorophyll photosensitizer

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The spectroscopic properties of a new chlorophyll derivate photosensitizer(CDP) are studied under the excitation wavelengths at 800 and 400 nm using femtosecond pulses from a Ti:sapphire laser.The damaging effect of CDP on the BEL-7402 cancer cells is also investigated upon two-photon illumination at 800 nm.The normalized fluorescence spectra of CDP in tetrahydrofuran(THF) show that two-photon and one-photon spectra have the same distributions and the same emission bands(675 nm).The life-times of two-and one-photon induced fluorescence of this molecule are of the order of 5.0 ns.By comparing the data it is shown that there is some difference between the two lifetimes,but the differ-ence is less than one nanosecond.The two-photon absorption cross section of the molecule is also measured at 800 nm and estimated as about σ′2 ≈ 31.5×10-50 cm4·s·photon-1.The results of two-photon photodynamic therapy(TPPDT) tests show that CDP can kill all of the tested cancer cells according to the usual Eosine assessment.Our results indicate that the two-photon-induced photophysical,photo-chemical and photosensitizing processes of CDP may be basically similar to those of one-photon ex-citation.These behaviors of the sample suggest that one may find other possible methods to estimate some photosensitizers’ effects in details such as their distribution in cells and the reactive targets of the sub-cellular parts of some tumor cells via two-photon excitation techniques.

  20. Electron correlations and two-photon states in polycyclic aromatic hydrocarbon molecules: A peculiar role of geometry

    Energy Technology Data Exchange (ETDEWEB)

    Aryanpour, Karan [Department of Physics, University of Arizona, Tucson, Arizona 85721 (United States); Shukla, Alok [Department of Physics, Indian Institute of Technology, Powai, Mumbai 400076 (India); Mazumdar, Sumit [Department of Physics, University of Arizona, Tucson, Arizona 85721 (United States); College of Optical Sciences, University of Arizona, Tucson, Arizona 85721 (United States)

    2014-03-14

    We present numerical studies of one- and two-photon excited states ordering in a number of polycyclic aromatic hydrocarbon molecules: coronene, hexa-peri-hexabenzocoronene, and circumcoronene, all possessing D{sub 6h} point group symmetry versus ovalene with D{sub 2h} symmetry, within the Pariser-Parr-Pople model of interacting π-electrons. The calculated energies of the two-photon states as well as their relative two-photon absorption cross-sections within the interacting model are qualitatively different from single-particle descriptions. More remarkably, a peculiar role of molecular geometry is found. The consequence of electron correlations is far stronger for ovalene, where the lowest spin-singlet two-photon state is a quantum superposition of pairs of lowest spin triplet states, as in the linear polyenes. The same is not true for D{sub 6h} group hydrocarbons. Our work indicates significant covalent character, in valence bond language, of the ground state, the lowest spin triplet state and a few of the lowest two-photon states in D{sub 2h} ovalene but not in those with D{sub 6h} symmetry.

  1. Electron correlations and two-photon states in polycyclic aromatic hydrocarbon molecules: A peculiar role of geometry

    Science.gov (United States)

    Aryanpour, Karan; Shukla, Alok; Mazumdar, Sumit

    2014-03-01

    We present numerical studies of one- and two-photon excited states ordering in a number of polycyclic aromatic hydrocarbon molecules: coronene, hexa-peri-hexabenzocoronene, and circumcoronene, all possessing D6h point group symmetry versus ovalene with D2h symmetry, within the Pariser-Parr-Pople model of interacting π-electrons. The calculated energies of the two-photon states as well as their relative two-photon absorption cross-sections within the interacting model are qualitatively different from single-particle descriptions. More remarkably, a peculiar role of molecular geometry is found. The consequence of electron correlations is far stronger for ovalene, where the lowest spin-singlet two-photon state is a quantum superposition of pairs of lowest spin triplet states, as in the linear polyenes. The same is not true for D6h group hydrocarbons. Our work indicates significant covalent character, in valence bond language, of the ground state, the lowest spin triplet state and a few of the lowest two-photon states in D2h ovalene but not in those with D6h symmetry.

  2. Organic nanostructure-based probes for two-photon imaging of mitochondria and microbes with emission between 430 nm and 640 nm.

    Science.gov (United States)

    Yang, Xinglong; Wang, Nuoxin; Zhang, Lingmin; Dai, Luru; Shao, Huawu; Jiang, Xingyu

    2017-04-06

    Multi-photon excitation and versatile fluorescent probes are in high need for biological imaging, since one probe can satisfy many needs as a biosensor. Herein we synthesize a series of two-photon excited probes based on tetraphenylethene (TPE) structures (TPE-Acr, TPE-Py, and TPE-Quino), which can image both mammalian cells and bacteria based on aggregation-induced emission (AIE) without washing them. Because of cationic moieties, the fluorescent molecules can aggregate into nanoscale fluorescent organic nanoscale dots to image mitochondria and bacteria with tunable emissions using both one-photon and two-photon excitation. Our research demonstrates that these AIE-dots expand the functions of luminescent organic dots to construct efficient fluorescent sensors applicable to both one-photon and two-photon excitation for bio-imaging of bacteria and mammalian cells.

  3. Internal conversions in Higgs decays to two photons

    OpenAIRE

    Firan, Ana; Stroynowski, Ryszard

    2007-01-01

    We evaluate the partial widths for internal conversions in the Higgs decays to two photons. For the Higgs masses of interest at LHC in the range of 100-150 GeV, the conversions to pairs of fermions represent significant fraction of Higgs decays.

  4. The 2 1Ag state of isolated cis-trans-1,3,5,7-octatetraene: two-color resonance enhanced two-photon ionization studies

    NARCIS (Netherlands)

    B.E. Kohler; T. Shaler; W.J. Buma

    1992-01-01

    Vibrationally resolved 1 1Ag2 1Ag excitation spectra and decay times for cis,trans-1,3,5,7-octatetraene seeded in a supersonic He expansion have been measured by two-color resonance enhanced two-photon ionization spectroscopy. The excitation energy of the 1 1Ag2 1Ag 0-0 band (29 035 cm-1 ) is ~6500

  5. Spectral characterization and unmixing of intrinsic contrast in intact normal and diseased gastric tissues using hyperspectral two-photon microscopy.

    Directory of Open Access Journals (Sweden)

    Lauren E Grosberg

    Full Text Available BACKGROUND: Living tissues contain a range of intrinsic fluorophores and sources of second harmonic generation which provide contrast that can be exploited for fresh tissue imaging. Microscopic imaging of fresh tissue samples can circumvent the cost and time associated with conventional histology. Further, intrinsic contrast can provide rich information about a tissue's composition, structure and function, and opens the potential for in-vivo imaging without the need for contrast agents. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we used hyperspectral two-photon microscopy to explore the characteristics of both normal and diseased gastrointestinal (GI tissues, relying only on their endogenous fluorescence and second harmonic generation to provide contrast. We obtained hyperspectral data at subcellular resolution by acquiring images over a range of two-photon excitation wavelengths, and found excitation spectral signatures of specific tissue types based on our ability to clearly visualize morphology. We present the two-photon excitation spectral properties of four major tissue types that are present throughout the GI tract: epithelium, lamina propria, collagen, and lymphatic tissue. Using these four excitation signatures as basis spectra, linear unmixing strategies were applied to hyperspectral data sets of both normal and neoplastic tissue acquired in the colon and small intestine. Our results show that hyperspectral unmixing with excitation spectra allows segmentation, showing promise for blind identification of tissue types within a field of view, analogous to specific staining in conventional histology. The intrinsic spectral signatures of these tissue types provide information relating to their biochemical composition. CONCLUSIONS/SIGNIFICANCE: These results suggest hyperspectral two-photon microscopy could provide an alternative to conventional histology either for in-situ imaging, or intraoperative 'instant histology' of fresh tissue

  6. Red autofluorescence of dental plaque bacteria

    NARCIS (Netherlands)

    van der Veen, M. H.; Thomas, R. Z.; Huysmans, M. C. D. N. J. M.; de Soet, J. J.

    2006-01-01

    Red autofluorescence of plaque and its relation to fluorescence of a single species in the biofilm was studied. Fluorescence images of non-disclosed and disclosed plaque of 28 first-year students were captured. The plaque samples were assessed by culture methods and studied for red autofluorescence.

  7. Fluorescence enhancement of asCP595 is due to consecutive absorbance of two photons

    Science.gov (United States)

    Savitsky, Alexander P.; Agranat, Michail B.; Lukyanov, Konstantin A.; Schuttrigkeit, Tanja; von Feilitzsch, Till; Kompa, Christian; Michel-Beyerle, Maria-Elisabeth

    2004-06-01

    Colored proteins are widely used as gene markers in biotechnology. Chromophores result from autocatalytic posttranslational reactions involving several amino acids. The protein asCP595 was isolated for the first time from the coral as a weakly fluorescent chromoprotein with a fluorescence maximum at 595 nm. Strong illumination in the blue wing of the low energy absorption band results in a superlinear increase of the fluorescence yield and shifts its fluorescence spectrum by about 10 nm to the red. Time resolved fluorescence measurements using excitation pulses with 10 ps duration revealed a multiexponential decay pattern with time constants in the range from 20 ps to 2.1 ns. The ratio of amplitudes related to the different time constants depends on the intensity of illumination favoring the ns component at high intensities. Transient absorption measurements using ultrashort excitation pulses (150 fs, 1 kHz repetition rate) did not reveal excited states with nanosecond lifetimes as observed in fluorescence upon excitation using 10 ps pulses. This observation leads to the notion that within 10 ps a second photon is absorbed by a state not yet populated within 150 fs. As a consequence we propose two different excited singlet states operative in asCP595, one with low fluorescence quantum yield peaking at 595 nm and one with high fluorescence quantum yield peaking at 605 nm which is populated via the consecutive absorption of two photons at high excitation intensities.

  8. A simple approach for measuring FRET in fluorescent biosensors using two-photon microscopy.

    Science.gov (United States)

    Day, Richard N; Tao, Wen; Dunn, Kenneth W

    2016-11-01

    Genetically encoded fluorescent protein (FP)-based biosensor probes are useful tools for monitoring cellular events in living cells and tissues. Because these probes were developed for one-photon excitation approaches, their broad two-photon excitation (2PE) and poorly understood photobleaching characteristics have made their implementation in studies using two-photon laser-scanning microscopy (TPLSM) challenging. Here we describe a protocol that simplifies the use of Förster resonance energy transfer (FRET)-based biosensors in TPLSM. First, the TPLSM system is evaluated and optimized using FRET standards expressed in living cells, which enables the determination of spectral bleed-through (SBT) and the confirmation of FRET measurements from the known standards. Next, we describe how to apply the approach experimentally using a modified version of the A kinase activity reporter (AKAR) protein kinase A (PKA) biosensor as an example-first in cells in culture and then in hepatocytes in the liver of living mice. The microscopic imaging can be accomplished in a day in laboratories that routinely use TPLSM.

  9. On the origin of large two-photon activity of DANS molecule.

    Science.gov (United States)

    Alam, Md Mehboob; Chattopadhyaya, Mausumi; Chakrabarti, Swapan

    2012-11-15

    In this work, using the quadratic response theory and two-state model approach, we have explained the origin of high two-photon activity and the corresponding solvent dependence of 4,4'-dimethyl-amino-nitro-stilbene (DANS) molecule. For this purpose, we have made two structural modifications in the DANS molecule (1) at the donor-acceptor part and (2) at the unsaturated bridge between the two rings and calculated the one- and two-photon (OP and TP) absorption parameters of all the systems in gas phase and in three different solvents, viz., MeCN, THF, and toluene. We found that the removal of donor-acceptor groups from the original DANS molecule vanishes the transition moment between the ground and excited states and also the corresponding dipole moment difference, and the saturation of the π-conjugation bridge between the two rings keeping the donor-acceptor groups intact causes a large decrease in the ground to excited state transition moment. These changes, in turn, decrease the overall TP activity of the molecules as compared to DANS. On the basis of our analysis, we have concluded that neither the donor-acceptor pair nor the π-conjugation bridge between the two, rather their cooperative involvement leads to a large overlap between the ground and virtual and also the virtual and charge-transfer states, which are eventually responsible for the very large TP activity of DANS.

  10. Two-photon absorption of Zn(II) octupolar molecules.

    Science.gov (United States)

    Mazzucato, Simone; Fortunati, Ilaria; Scolaro, Sara; Zerbetto, Michele; Ferrante, Camilla; Signorini, Raffaella; Pedron, Danilo; Bozio, Renato; Locatelli, Danika; Righetto, Stefania; Roberto, Dominique; Ugo, Renato; Abbotto, Alessandro; Archetti, Graziano; Beverina, Luca; Ghezzi, Sergio

    2007-06-21

    In this work we present an investigation of the non-linear optical (NLO) properties of two octupolar chromophores: [Zn(4,4'-bis(dibutylaminostyryl)-[2,2']-bipyridine)(3)](2+) and [Zn(4,4'-bis((E)-2-(N-(TEG)pyrrol-2-yl)vinyl)-[2,2']-bipyridine)(3)](2+) with Zn(ii) as the coordination center, using two-photon emission technique (TPE) in fs-pulse temporal regime. Compared to the free ligands, our results do not show a net increase in the two-photon absorption (TPA) cross-section for the octupolar complexes, once normalized to the ligand unit. This is in partial disagreement with a previous theoretical study investigating the first molecule where a significant increase of the TPA cross-section was predicted (X. J. Liu, et al., J. Chem. Phys., 2004, 120, 11 493).

  11. Synthesizing arbitrary two-photon polarization mixed states

    CERN Document Server

    Wei, T C; Branning, D; Goldbart, P M; James, D F V; Jeffrey, E; Kwiat, P G; Mukhopadhyay, S; Peters, N A; Wei, Tzu-Chieh; Altepeter, Joseph B.; Branning, David; Goldbart, Paul M.; Jeffrey, Evan; Kwiat, Paul G.; Mukhopadhyay, Swagatam; Peters, Nicholas A.

    2005-01-01

    Two methods for creating arbitrary two-photon polarization pure states are introduced. Based on these, four schemes for creating two-photon polarization mixed states are proposed and analyzed. The first two schemes can synthesize completely arbitrary two-qubit mixed states, i.e., control all 15 free parameters: Scheme I requires several sets of crystals, while Scheme II requires only a single set, but relies on decohering the pump beam. Additionally, we describe two further schemes which are much easier to implement. Although the total capability of these is still being studied, we show that they can synthesize all two-qubit Werner states, maximally entangled mixed states, Collins-Gisin states, and arbitrary Bell-diagonal states.

  12. Direct frequency comb two-photon laser cooling and trapping

    Science.gov (United States)

    Jayich, Andrew; Long, Xueping; Campbell, Wesley C.

    2016-05-01

    Generating and manipulating high energy photons for spectroscopy on electric dipole transitions of atoms and molecules with deeply bound valence electrons is difficult. Further, laser cooling of such species is even more challenging for lack of laser power. A possible solution is to drive two-photon transitions. This may alleviate the photon energy problem and open the door to cold, trapped samples of highly desirable species with tightly bound electrons. We perform a proof of principle experiment with rubidium by driving a two-photon transition with an optical frequency comb. We perform optical cooling and extend this technique to trapping, where we are able to make a magneto-optical trap in one dimension. This work is supported by the National Science Foundation CAREER program.

  13. High-order dispersion effects in two-photon interference

    CERN Document Server

    Mazzotta, Z; Cipriani, D; Olivares, S; Paris, M G A

    2016-01-01

    Two-photon interference and Hong-Ou-Mandel (HOM) effect are relevant tools for quantum metrology and quantum information processing. In optical coherence tomography, HOM effect is exploited to achieve high-resolution measurements with the width of the HOM dip being the main parameter. On the other hand, applications like dense coding require high-visibility performances. Here we address high-order dispersion effects in two-photon interference and study, theoretically and experimentally, the dependence of the visibility and the width of the HOM dip on both the pump spectrum and the downconverted photon spectrum. In particular, a spatial light modulator is exploited to experimentally introduce and manipulate a custom phase function to simulate the high-order dispersion effects.

  14. Two-photon interference from two blinking quantum emitters

    Science.gov (United States)

    Jöns, Klaus D.; Stensson, Katarina; Reindl, Marcus; Swillo, Marcin; Huo, Yongheng; Zwiller, Val; Rastelli, Armando; Trotta, Rinaldo; Björk, Gunnar

    2017-08-01

    We investigate the effect of blinking on the two-photon interference measurement from two independent quantum emitters. We find that blinking significantly alters the statistics in the Hong-Ou-Mandel second-order intensity correlation function g(2 )(τ ) and the outcome of two-photon interference measurements performed with independent quantum emitters. We theoretically demonstrate that the presence of blinking can be experimentally recognized by a deviation from the gD(2 )(0 ) =0.5 value when distinguishable photons from two emitters impinge on a beam splitter. Our findings explain the significant differences between linear losses and blinking for correlation measurements between independent sources and are experimentally verified using a parametric down-conversion photon-pair source. We show that blinking imposes a mandatory cross-check measurement to correctly estimate the degree of indistinguishability of photons emitted by independent quantum emitters.

  15. Two-photon interaction between trapped ions and cavity fields

    CERN Document Server

    Semião, F L

    2006-01-01

    In this paper, we generalize the ordinary two-photon Jaynes-Cummings model (TPJCM) by considering the atom (or ion) to be trapped in a simple harmonic well. A typical setup would be an optical cavity containing a single ion in a Paul trap. Due to the inclusion of atomic vibrational motion, the atom-field coupling becomes highly nonlinear what brings out quite different behaviors for the system dynamics when compared to the ordinary TPJCM. In particular, we derive an effective two-photon Hamiltonian with dependence on the number operator of the ion's center-of-mass motion. This dependence occurs both in the cavity induced Stark-shifs and in the ion-field coupling, and its role in the dynamics is illustrated by showing the time evolution of the probability of occupation of the electronic levels for simple initial preparations of the state of the system.

  16. Two-photon-induced cycloreversion reaction of chalcone photodimers

    Science.gov (United States)

    Träger, J.; Härtner, S.; Heinzer, J.; Kim, H.-C.; Hampp, N.

    2008-04-01

    The photocleavage reaction of chalcone photodimers has been studied using a two-photon process. For this purpose, a novel chalcone dimer has been synthesized as a low molecular weight model substance for polymer bound chalcones and its photochemistry triggered by two-photon-absorption (2PA) has been investigated using a pulsed frequency-doubled Nd:YAG-laser. The 2PA-induced cycloreversion reaction selectively leads to the cleavage of the chalcone photodimers resulting in the formation of monomeric chalcone molecules. Hence, as an application chalcones can be used as a photosensitive linker which can be cleaved beyond an UV-absorbing barrier. The 2PA cross section of the chalcone photodimer was determined to be of 1.1 × 10 -49 cm 4 s photon -1 (11 GM).

  17. Two-photon or higher-order absorbing optical materials and methods of use

    Science.gov (United States)

    Marder, Seth (Inventor); Perry, Joseph (Inventor)

    2012-01-01

    Compositions capable of simultaneous two-photon absorption and higher order absorptivities are provided. Compounds having a donor-pi-donor or acceptor-pi-acceptor structure are of particular interest, where the donor is an electron donating group, acceptor is an electron accepting group, and pi is a pi bridge linking the donor and/or acceptor groups. The pi bridge may additionally be substituted with electron donating or withdrawing groups to alter the absorptive wavelength of the structure. Also disclosed are methods of generating an excited state of such compounds through optical stimulation with light using simultaneous absorption of photons of energies individually insufficient to achieve an excited state of the compound, but capable of doing so upon simultaneous absorption of two or more such photons. Applications employing such methods are also provided, including controlled polymerization achieved through focusing of the light source(s) used.

  18. Two-photon imaging of lymphoma cells targeted by gold nanoparticles

    Institute of Scientific and Technical Information of China (English)

    Xiaochao Qu; Jing Wang; Cuiping Yao; Zhenxi Zhang

    2008-01-01

    Gold nanoparticles (NPs) have highly efficient multi-photon-induced luminescence. In this paper, we record the two-photon images of gold NPs, lymphoma cell line Karpas 299, and Karpas 299 incubated with 30-nm-diameter gold NPs and ACT-1 antibody conjugates (Au30-ACT-1 conjugates) by using a multi-photon microscopy system. Due to the specific conjugation of ACT-1 antibody and celt membrane receptor CD25, gold NPs are only bound to the surface of cell membrane of Karpas 299. The luminescence intensity of gold NPs is higher than that of cells at 750-nm laser excitation. By comparing the images of Karpas 299 cells incubated with and without gold NPs, it is found that by means of gold NPs, we can get clear cell images with lower excitation power. Their excellent optical and chemical properties make gold NPs an attractive contrast agent for cellular imaging.

  19. Autofluorescence of human cells in vitro as a biomarker of their metabolic activity

    Science.gov (United States)

    Dobrzyńska, Monika; Stepińska, Małgorzata; Lewandowski, Rafał; Gietka, Andrzej; Łapiński, Mariusz P.; Trafny, ElŻbieta A.

    2016-12-01

    Autofluorescence (AF) is the natural emission of light by intrinsic fluorophores. Oxidized mitochondrial flavins, lipofuscin and reduced nicotinamideadenine dinucleotide phosphate (NAD(P)H) are the main sources of the autofluorescence in cells upon excitation with visible light. The aim of the study was to investigate changes in the metabolism of four cell lines by monitoring their autofluorescence with a microplate reader. Autofluorescence intensities of cells were collected at two wavelengths for the excitation and fluorescence emission: for endogenous NAD(P)H at 366/450 nm, for the oxidized flavoproteins and lipofuscin at 460/540 nm. Human mesenchymal stem cells (hMSC), epithelial cells from mammary gland (MCF 10A), breast ductal carcinoma (T-47D) prostate carcinoma (DU-145) were observed daily for 16 days. The level of NAD(P)H autofluorescence did not differ among the cell lines investigated. The significant increase in oxidized flavoproteins fluorescence intensity was recorded for hMSC and ranged from 140 to 175% of control. During 28 days differentiation process, the NAD(P)H, FAD and lipofuscin fluorescence intensities were recorded every day, and the redox ratio was then calculated. The redox ratio gradually decreased during the last eight days of osteogenesis and adipogenesis. Therefore, in our opinion the NAD(P)H, FAD, and lipofuscin fluorescence emission at the wavelengths selected are the optimal parameters to be collected during the differentiation process in order to monitor the metabolism of hMSC undergoing structural and morphological changes.

  20. Margins of oral leukoplakia: autofluorescence and histopathology.

    Science.gov (United States)

    Elvers, D; Braunschweig, T; Hilgers, R-D; Ghassemi, A; Möhlhenrich, S C; Hölzle, F; Gerressen, M; Modabber, A

    2015-02-01

    Autofluorescence devices are widely used to examine oral lesions. The aim of this study was to see whether there were any signs of dysplasia, parakeratosis, or mucosal inflammation in the borders of homogeneous oral leukoplakia using autofluorescence, and we also compared clinically visible extensions with those detected by autofluorescence. Twenty patients with 26 homogeneous areas of oral leukoplakia were included in the study. After the clinically visible extensions of the lesion had been marked, we took a photograph through the autofluorescence device, which showed both borders in one picture. We then used photo-editing software to measure the size of the area of leukoplakia together with the area with loss of autofluorescence. We took 3 punch biopsy specimens: one from the leukoplakia, one 2.5mm from its marked borders, and one from healthy mucosa. Seventy-eight biopsy specimens were examined by an experienced pathologist, and 95% CI calculated to assess the amount of parakeratosis. Spearman's rank correlation was used to assess the association with mucosal inflammation. Ten areas of leukoplakia were surrounded by normal green autofluorescence, and 16 were consistent with loss of autofluorescence with a mean size of 66%, which exceeded the clinically visible size of the area of leukoplakia. We calculated that there was a strong association between these entities and their surrounding areas, with loss of autofluorescence for parakeratosis. Some leukoplakias showed clinically invisible extensions during histopathological examination and autofluorescence. The technique described enables clinicians to measure the extent of these lesions beyond their visible margins. We found no dysplasia, which emphasises that autofluorescence detects non-dysplastic lesions caused by mucosal inflammation and parakeratosis.

  1. Two-photon production of charged pion and kaon pairs

    CERN Document Server

    Dominick, J; Sanghera, S; Shelkov, V; Skwarnicki, T; Stroynowski, R; Volobuev, I P; Wei, G; Zadorozhny, P; Artuso, M; Goldberg, M; He, D; Horwitz, N; Kennett, R; Mountain, R; Moneti, G C; Muheim, F; Mukhin, Y; Playfer, S; Rozen, Y; Stone, S; Thulasidas, M; Vasseur, G; Zhu, G; Bartelt, J; Csorna, S E; Egyed, Z; Jain, V; Kinoshita, K; Edwards, K W; Ogg, M; Britton, D I; Hyatt, E R F; MacFarlane, D B; Patel, P M; Akerib, D S; Barish, B C; Chadha, M; Chan, S; Cowen, D F; Eigen, G; Miller, J S; O'Grady, C; Urheim, J; Weinstein, A J; Acosta, D; Athanas, M; Masek, G E; Paar, H P; Sivertz, M; Gronberg, J B; Kutschke, R; Menary, S R; Morrison, R J; Nakanishi, S; Nelson, H N; Nelson, T K; Qiao, C; Richman, J D; Ryd, A; Tajima, H; Sperka, D; Witherell, M S; Procario, M; Balest, R; Cho, K; Daoudi, M; Ford, W T; Johnson, D R; Lingel, K; Lohner, M; Rankin, P; Smith, J G; Alexander, J P; Bebek, C; Berkelman, K; Bloom, K; Browder, T E; Cassel, David G; Cho, H A; Coffman, D M; Drell, P S; Ehrlich, R; Gaidarev, P B; Galik, R S; García-Sciveres, M; Geiser, B; Gittelman, B; Gray, S W; Hartill, D L; Heltsley, B K; Jones, C D; Jones, S L; Kandaswamy, J; Katayama, N; Kim, P C; Kreinick, D L; Ludwig, G S; Masui, J; Mevissen, J; Mistry, N B; Ng, C R; Nordberg, E; Patterson, J R; Peterson, D; Riley, D; Salman, S; Sapper, M; Würthwein, F; Avery, P; Freyberger, A P; Rodríguez, J; Stephens, R; Yang, S; Yelton, J; Cinabro, D; Henderson, S; Liu, T; Saulnier, M; Wilson, R; Yamamoto, H; Bergfeld, T; Eisenstein, B I; Gollin, G; Ong, B; Palmer, M; Selen, M; Thaler, J J; Sadoff, A J; Ammar, R; Ball, S; Baringer, P; Bean, A; Besson, D; Coppage, D; Copty, N K; Davis, R; Hancock, N; Kelly, M; Kwak, N; Lam, H; Kubota, Y; Lattery, M; Nelson, J K; Patton, S; Perticone, D; Poling, R A; Savinov, V; Schrenk, S; Wang, R; Alam, M S; Kim, I J; Nemati, B; O'Neill, J J; Severini, H; Sun, C R; Zoeller, M M; Crawford, G; Daubenmier, C M; Fulton, R; Fujino, D; Gan, K K; Honscheid, K; Kagan, H; Kass, R; Lee, J; Malchow, R L; Skovpen, Y; Sung, M; White, C; Butler, F; Fu, X; Kalbfleisch, G R; Ross, W R; Skubic, P L; Snow, J; Wang, P L; Wood, M; Brown, D N; Fast, J; McIlwain, R L; Miao, T; Miller, D H; Modesitt, M; Payne, D; Shibata, E I; Shipsey, I P J; Wang Pei Ning; Battle, M; Ernst, J; Kwon, Y; Roberts, S; Thorndike, E H; Wang, C H

    1994-01-01

    A measurement of the cross section for the combined two-photon production of charged pion and kaon pairs is performed using 1.2~\\rm fb^{-1} of data collected by the CLEO II detector at the Cornell Electron Storage Ring. The cross section is measured at invariant masses of the two-photon system between 1.5 and 5.0~GeV/c^2, and at scattering angles more than 53^\\circ away from the \\gamma\\gamma collision axis in the \\gamma\\gamma center-of-mass frame. The large background of leptonic events is suppressed by utilizing the CsI calorimeter in conjunction with the muon chamber system. The reported cross section is compared with leading order QCD models as well as previous experiments. In particular, although the functional dependence of the measured cross section disagrees with leading order QCD at small values of the two-photon invariant mass, the data show a transition to perturbative behavior at an invariant mass of approximately 2.5~GeV/c^2. hardcopies with figures can be obtained by writing to to: Pam Morehouse ...

  2. Two photon exchange in elastic electron-nucleon scattering

    Energy Technology Data Exchange (ETDEWEB)

    Peter Blunden; Wolodymyr Melnitchouk; John Tjon

    2005-06-01

    A detailed study of two-photon exchange in unpolarized and polarized elastic electron-nucleon scattering is presented, taking particular account of nucleon finite size effects. Contributions from nucleon elastic intermediate states are found to have a strong angular dependence, which leads to a partial resolution of the discrepancy between the Rosenbluth and polarization transfer measurements of the proton electric to magnetic form factor ratio. The two-photon exchange contribution to the longitudinal polarization transfer ratio P{sub L} is small, whereas the contribution to the transverse polarization transfer ratio P{sub T} is enhanced at backward angles by several percent, increasing with Q{sup 2}. This gives rise to a several percent enhancement of the polarization transfer ratio P{sub T}/P{sub l} at large Q{sup 2} and backward angles. We compare the two-photon exchange effects with data on the ratio of e{sup +p} to e{sup -p} cross sections, which is predicted to be enhanced at backward angles. Finally, we evaluate the corrections to the form factors of the neutron, and estimate the elastic intermediate state contribution to the {sup 3}He form factors.

  3. Recent two-photon physics results from ARGUS

    Science.gov (United States)

    Živko Representing Argus Collaboration, Tomi

    1995-07-01

    Two photon production of π+π+π0π-π-, K+K-π+π-, K+K-π+π0π-, π+π0π-, and π+π- has been studied using the ARGUS detector at the e+e- storage ring DORIS II at DESY. A partial wave analysis was performed on the five-pion and three-pion final states. In the reaction γγ→ωρ0 is showed that the partial-wave with spin and parity (JP,Jz)=(2+,±2) dominates. The cross section and angular distributions of the reaction γγ→φρ0→K+K-π+π- were measured for the first time. The production of the vector-meson pair φω is observed in the two-photon reaction γγ→K+K-π+π0π-. The two-photon width of the tensor meson a2(1320) was measured in the decay channel π+π0π-. An upper limit, significantly lower than indicated by previous experiments was set on the radiative width of the π2(1670) meson. An upper limit was set on the radiative width of the f0(975)in the decay channel π+π-.

  4. Two-Photon Absorption in Organometallic Bromide Perovskites

    KAUST Repository

    Walters, Grant

    2015-07-21

    Organometallic trihalide perovskites are solution processed semiconductors that have made great strides in third generation thin film light harvesting and light emitting optoelectronic devices. Recently it has been demonstrated that large, high purity single crystals of these perovskites can be synthesized from the solution phase. These crystals’ large dimensions, clean bandgap, and solid-state order, have provided us with a suitable medium to observe and quantify two-photon absorption in perovskites. When CH3NH3PbBr3 single crystals are pumped with intense 800 nm light, we observe band-to-band photoluminescence at 572 nm, indicative of two-photon absorption. We report the nonlinear absorption coefficient of CH3NH3PbBr3 perovskites to be 8.6 cm GW-1 at 800 nm, comparable to epitaxial single crystal semiconductors of similar bandgap. We have leveraged this nonlinear process to electrically autocorrelate a 100 fs pulsed laser using a two-photon perovskite photodetector. This work demonstrates the viability of organometallic trihalide perovskites as a convenient and low-cost nonlinear absorber for applications in ultrafast photonics.

  5. Exploring control parameters of two photon processes in solutions

    Indian Academy of Sciences (India)

    Debabrata Goswami; Amit Nag

    2012-01-01

    Two-photon microscopy depends extensively on the two-photon absorption cross-sections of biologically relevant chromophores. High repetition rate (HRR) lasers are essential in multiphoton microscopy for generating satisfactory signal to noise at low average powers. However, HRR lasers generate thermal distortions in samples even with the slightest single photon absorption. We use an optical chopper with HRR lasers to intermittently `blank’ irradiation and effectively minimize thermal effects to result in a femtosecond z-scan setup that precisely measures the two-photon absorption (TPA) cross-sections of chromophores. Though several experimental factors impact such TPA measurements, a systematic effort to modulate and influence TPA characteristics is yet to evolve. Here, we present the effect of several control parameters on the TPA process that are independent of chromophore characteristics for femtosecond laser pulse based measurements; and demonstrate how the femtosecond laser pulse repetition rate, chromophore environment and incident laser polarization can become effective control parameters for such nonlinear optical properties.

  6. Two-Photon Absorption in Organometallic Bromide Perovskites.

    Science.gov (United States)

    Walters, Grant; Sutherland, Brandon R; Hoogland, Sjoerd; Shi, Dong; Comin, Riccardo; Sellan, Daniel P; Bakr, Osman M; Sargent, Edward H

    2015-09-22

    Organometallic trihalide perovskites are solution-processed semiconductors that have made great strides in third-generation thin film light-harvesting and light-emitting optoelectronic devices. Recently, it has been demonstrated that large, high-purity single crystals of these perovskites can be synthesized from the solution phase. These crystals' large dimensions, clean bandgap, and solid-state order have provided us with a suitable medium to observe and quantify two-photon absorption in perovskites. When CH3NH3PbBr3 single crystals are pumped with intense 800 nm light, we observe band-to-band photoluminescence at 572 nm, indicative of two-photon absorption. We report the nonlinear absorption coefficient of CH3NH3PbBr3 perovskites to be 8.6 cm GW(-1) at 800 nm, comparable to epitaxial single-crystal semiconductors of similar bandgap. We have leveraged this nonlinear process to electrically autocorrelate a 100 fs pulsed laser using a two-photon perovskite photodetector. This work demonstrates the viability of organometallic trihalide perovskites as a convenient and low-cost nonlinear absorber for applications in ultrafast photonics.

  7. Ultrafast relaxation kinetics of the dark S{sub 1} state in all-trans-{beta}-carotene explored by one- and two-photon pump-probe spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Kosumi, Daisuke, E-mail: kosumi@sci.osaka-cu.ac.jp [Department of Physics, Graduate School of Science, Tohoku University, 6-3 Aramaki-Aza-Aoba, Sendai 980-8578 (Japan); Abe, Kenta; Karasawa, Hiroshi [Department of Physics, Graduate School of Science, Tohoku University, 6-3 Aramaki-Aza-Aoba, Sendai 980-8578 (Japan); Fujiwara, Masazumi [Department of Physics, Graduate School of Science, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585 (Japan); Cogdell, Richard J. [Glasgow Biomedical Research Center, University of Glasgow, 120 University Place, Glasgow G12 8TA, Scotland (United Kingdom); Hashimoto, Hideki [Department of Physics, Graduate School of Science, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585 (Japan); JST/CREST, 4-1-8 Hon-chou, Kawaguchi, Saitama 332-0012 (Japan); Yoshizawa, Masayuki [Department of Physics, Graduate School of Science, Tohoku University, 6-3 Aramaki-Aza-Aoba, Sendai 980-8578 (Japan); JST/CREST, 4-1-8 Hon-chou, Kawaguchi, Saitama 332-0012 (Japan)

    2010-07-19

    Femtosecond one- and two-photon pump-probe dispersive spectroscopic measurements have been applied to the investigation of the vibrational relaxation kinetics of the dark S{sub 1} (2{sup 1}A{sub g}{sup -}) state in {beta}-carotene, combining a higher sensitive detection system with tunable visible and infrared excitation pulses. The two-photon excitation measurements enable the preferential detection of the dark S{sub 1} state. The tunable infrared excitation pulses allowed selective excitation to a different vibrational level of S{sub 1}. The S{sub 1} dynamics at early delay times depend strongly on excitation energy. A dependence of the initial S{sub 1} dynamics on excitation energy is discussed in term of the vibrational relaxation of S{sub 1}.

  8. Polarization control efficiency manipulation in resonance-mediated two-photon absorption by femtosecond spectral frequency modulation

    Science.gov (United States)

    Yao, Yunhua; Cheng, Wenjing; Zheng, Ye; Xu, Cheng; Liu, Pei; Jia, Tianqing; Qiu, Jianrong; Sun, Zhenrong; Zhang, Shian

    2017-04-01

    The femtosecond laser polarization modulation is considered as a very simple and efficient method to control the multi-photon absorption process. In this work, we theoretically and experimentally show that the polarization control efficiency in the resonance-mediated two-photon absorption can be artificially manipulated by modulating the femtosecond spectral frequency components. We theoretically demonstrate that the on- and near-resonant parts in the resonance-mediated two-photon absorption process depend on the different femtosecond spectral frequency components, and therefore their contributions in the whole excitation process can be controlled by properly designing the femtosecond spectral frequency components. The near-resonant two-photon absorption is correlated with the femtosecond laser polarization while the on-resonant two-photon absorption is independent of it, and thus the polarization control efficiency in the resonance-mediated two-photon absorption can be manipulated by the femtosecond spectral frequency modulation. We experimentally verify these theoretical results by performing the laser polarization control experiment in the Dy3+-doped glass sample under the modulated femtosecond spectral frequency components, and the experimental results show that the polarization control efficiency can be increased when the central spectral frequency components are cut off, while it is decreased when both the low and high spectral frequency components are cut off, which is in good agreement with the theoretical predictions. Our works can provide a feasible pathway to understand and control the resonance-mediated multi-photon absorption process under the femtosecond laser field excitation, and also may open a new opportunity to the related application areas.

  9. (Un)determined finite regularization dependent quantum corrections: the Higgs decay into two photons and the two photon scattering examples

    CERN Document Server

    Cherchiglia, A L; Nemes, M C; Sampaio, Marcos

    2012-01-01

    We investigate the appearance of arbitrary, regularization dependent parameters introduced by divergent integrals in two a priori finite but superficially divergent amplitudes: the Higgs decay into two photons and the two photon scattering. We use a general parametrization of ultraviolet divergences which explicitates such ambiguities. Thus we separate in a consistent way using Implicit Regularization the divergent, finite and regularization dependent parts of the amplitudes which in turn are written as surface terms. We find that, although finite, these amplitudes are ambiguous before the imposition of physical conditions namely momentum routing invariance in the loops of Feynman diagrams. In the examples we study momentum routing invariance turns out to be equivalent to gauge invariance. We also discuss the results obtained by different regularizations and show how they can be reproduced within our framework allowing for a clear view on the origin of regularization ambiguities.

  10. 540--900 nm photodissociation of 300 K NCNO: One- and two-photon processes

    Energy Technology Data Exchange (ETDEWEB)

    Nadler, I.; Pfab, J.; Reisler, H.; Wittig, C.

    1984-07-15

    The laser photodissociation of 300 K NCNO throughout the region 540--900 nm is reported, and both 1- and 2-photon processes are discussed. By monitoring CN fragments produced via the 1-photon process, we show that with photolysis wavelengths >592 nm, dissociation occurs predominantly by exciting NCNO ''hot bands.'' At shorter photolysis wavelengths, dissociation from the ground vibrational state of NCNO is observed as well, but the contributions from hot bands are still manifest in high CN rotational levels which are energetically inaccessible from the ground state (D/sub 0/ = 48.8 kcal mol/sup -1/). Energy distributions in the CN fragments were determined for excess energies up to 1800 cm/sup -1/, and are in agreement with phase space theory calculations and a vibrational predissociation mechanism. In addition, throughout the region 620--900 nm, stepwise two-photon photodissociation proceeds using the A /sup 1/A'' state as a gateway, and results in rotationally and vibrationally ''hot'' CN fragments. The hot CN fragment yield vs photolysis wavelength shows peaks which correspond exactly to peaks in the NCNO absorption spectrum, allowing us to obtain high resolution spectra of the A /sup 1/A''reverse arrow X /sup 1/A' absorption system. The one- and two-photon processes are in competition, and the latter disappears at wavelengths where one-photon photodissociation of NCNO via its ground vibrational level sets in. The nature of the electronic states involved in the one- and two-photon processes is also discussed.

  11. Relativistic (Z alpha)^2-Corrections and Leading Quantum Electrodynamic Corrections to the Two-Photon Decay Rate of Ionic States

    CERN Document Server

    Wundt, B J; 10.1103/PhysRevA.80.022505

    2009-01-01

    We calculate the relativistic corrections of relative order (Z alpha)^2$ to the two-photon decay rate of higher excited S and D states in ionic atomic systems, and we also evaluate the leading radiative corrections of relative order alpha (Z alpha)^2 ln[(Z alpha)^(-2)]. We thus complete the theory of the two-photon decay rates up to relative order alpha^3 ln(alpha). An approach inspired by nonrelativistic quantum electrodynamics is used. We find that the corrections of relative order (Z alpha)^2 to the two-photon decay are given by the zitterbewegung, the spin-orbit coupling and by relativistic corrections to the electron mass, and by quadrupole interactions. We show that all corrections are separately gauge-invariant with respect to a "hybrid" transformation from velocity to length gauge, where the gauge transformation of the wave function is neglected. The corrections are evaluated for the two-photon decay from 2S, 3S, 3D, and 4S states in one-electron (hydrogenlike) systems, with 1S and 2S final states.

  12. Stepwise Two-Photon-Gated Photochemical Reaction in Photochromic [2.2]Paracyclophane-Bridged Bis(imidazole dimer).

    Science.gov (United States)

    Mutoh, Katsuya; Nakagawa, Yuki; Sakamoto, Akira; Kobayashi, Yoichi; Abe, Jiro

    2015-05-06

    Stepwise two-photon processes not only have great potential for efficient light harvesting but also can provide valuable insights into novel photochemical sciences. Here we have designed a [2.2]paracyclophane-bridged bis(imidazole dimer), a molecule that is composed of two photochromic units and absorbs two photons in a stepwise manner. The absorption of the first photon leads to the formation of a short-lived biradical species (half-life = 88 ms at 298 K), while the absorption of the additional photon by the biradical species triggers a subsequent photochromic reaction to afford a long-lived quinoid species. The short-lived biradical species and the long-lived quinoid species display significantly different absorption spectra and rates of the thermal back-reaction. The stepwise two-photon excitation process in this photochromic system can be initiated even by incoherent continuous-wave light irradiation, indicating that this two-photon reaction is highly efficient. Our molecule based on the bridged bis(imidazole dimer) unit should be a good candidate for multiphoton-gated optical materials.

  13. Two-photon absorption-induced photoacoustic imaging of Rhodamine B dyed polyethylene spheres using a femtosecond laser.

    Science.gov (United States)

    Langer, Gregor; Bouchal, Klaus-Dieter; Grün, Hubert; Burgholzer, Peter; Berer, Thomas

    2013-09-23

    In the present paper we demonstrate the possibility to image dyed solids, i.e. Rhodamine B dyed polyethylene spheres, by means of two-photon absorption-induced photoacoustic scanning microscopy. A two-photon luminescence image is recorded simultaneously with the photoacoustic image and we show that location and size of the photoacoustic and luminescence image match. In the experiments photoacoustic signals and luminescence signals are generated by pulses from a femtosecond laser. Photoacoustic signals are acquired with a hydrophone; luminescence signals with a spectrometer or an avalanche photo diode. In addition we derive the expected dependencies between excitation intensity and photoacoustic signal for single-photon absorption, two-photon absorption and for the combination of both. In order to verify our setup and evaluation method the theoretical predictions are compared with experimental results for liquid and solid specimens, i.e. a carbon fiber, Rhodamine B solution, silicon, and Rhodamine B dyed microspheres. The results suggest that the photoacoustic signals from the Rhodamine B dyed microspheres do indeed stem from two-photon absorption.

  14. Resolution of overlapping skin auto-fluorescence for development of non-invasive applications

    Science.gov (United States)

    Su, Yu-Zheng; Lin, Li-Wu; Chen, Chuen-Yau; Hung, Min-Wei; Huang, Kuo-Cheng

    2010-08-01

    Skin auto-fluorescence spectra can provide useful biological information, but the obtained spectrum is overlapped and is difficult to distinguish each contributed component. We applied the genetic algorithm to decompose the overlapping spectrum. First, we simulate the overlapping spectral to confirm our feasible algorithm. The skin auto-fluorescence spectra were obtained from the normal human skin with 337 nm excitation light source. The nicotinamide adenine dinucleotid (NADH) and flavin adenine dinucleotide (FAD) are accurately decomposed and demonstrated. The developed algorithm can be widely applied to achieve qualitative and quantitative analysis for overlapping spectra.

  15. Generation of molecular diffuse-band stimulated radiation through multiple excitation mechanism in potassium molecule-atom system

    Institute of Scientific and Technical Information of China (English)

    贾锁堂; 秦莉娟; 钱祖良; 王祖赓; 王钢; 周国生

    1997-01-01

    Population reduction effect generated in two-photon resonant excitation process of potassium atoms in a potassium molecule-atom system is theoretically investigated The obtained result is compared with the relevant experimental ones in the process of two-photon resonant excitation of potassium molecules and in the process of two-photon mixed excitation of potassium molecule-atoms.

  16. Two-photon imaging and analysis of neural network dynamics

    Science.gov (United States)

    Lütcke, Henry; Helmchen, Fritjof

    2011-08-01

    The glow of a starry night sky, the smell of a freshly brewed cup of coffee or the sound of ocean waves breaking on the beach are representations of the physical world that have been created by the dynamic interactions of thousands of neurons in our brains. How the brain mediates perceptions, creates thoughts, stores memories and initiates actions remains one of the most profound puzzles in biology, if not all of science. A key to a mechanistic understanding of how the nervous system works is the ability to measure and analyze the dynamics of neuronal networks in the living organism in the context of sensory stimulation and behavior. Dynamic brain properties have been fairly well characterized on the microscopic level of individual neurons and on the macroscopic level of whole brain areas largely with the help of various electrophysiological techniques. However, our understanding of the mesoscopic level comprising local populations of hundreds to thousands of neurons (so-called 'microcircuits') remains comparably poor. Predominantly, this has been due to the technical difficulties involved in recording from large networks of neurons with single-cell spatial resolution and near-millisecond temporal resolution in the brain of living animals. In recent years, two-photon microscopy has emerged as a technique which meets many of these requirements and thus has become the method of choice for the interrogation of local neural circuits. Here, we review the state-of-research in the field of two-photon imaging of neuronal populations, covering the topics of microscope technology, suitable fluorescent indicator dyes, staining techniques, and in particular analysis techniques for extracting relevant information from the fluorescence data. We expect that functional analysis of neural networks using two-photon imaging will help to decipher fundamental operational principles of neural microcircuits.

  17. Two-photon imaging and analysis of neural network dynamics

    Energy Technology Data Exchange (ETDEWEB)

    Luetcke, Henry; Helmchen, Fritjof [Brain Research Institute, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich (Switzerland)

    2011-08-15

    The glow of a starry night sky, the smell of a freshly brewed cup of coffee or the sound of ocean waves breaking on the beach are representations of the physical world that have been created by the dynamic interactions of thousands of neurons in our brains. How the brain mediates perceptions, creates thoughts, stores memories and initiates actions remains one of the most profound puzzles in biology, if not all of science. A key to a mechanistic understanding of how the nervous system works is the ability to measure and analyze the dynamics of neuronal networks in the living organism in the context of sensory stimulation and behavior. Dynamic brain properties have been fairly well characterized on the microscopic level of individual neurons and on the macroscopic level of whole brain areas largely with the help of various electrophysiological techniques. However, our understanding of the mesoscopic level comprising local populations of hundreds to thousands of neurons (so-called 'microcircuits') remains comparably poor. Predominantly, this has been due to the technical difficulties involved in recording from large networks of neurons with single-cell spatial resolution and near-millisecond temporal resolution in the brain of living animals. In recent years, two-photon microscopy has emerged as a technique which meets many of these requirements and thus has become the method of choice for the interrogation of local neural circuits. Here, we review the state-of-research in the field of two-photon imaging of neuronal populations, covering the topics of microscope technology, suitable fluorescent indicator dyes, staining techniques, and in particular analysis techniques for extracting relevant information from the fluorescence data. We expect that functional analysis of neural networks using two-photon imaging will help to decipher fundamental operational principles of neural microcircuits.

  18. Two-photon quantum interference in plasmonics: theory and applications.

    Science.gov (United States)

    Gupta, S Dutta; Agarwal, G S

    2014-01-15

    We report perfect two-photon quantum interference with near-unity visibility in a resonant tunneling plasmonic structure in folded Kretschmann geometry. This is despite absorption-induced loss of unitarity in plasmonic systems. The effect is traced to perfect destructive interference between the squares of amplitude reflection and transmission coefficients. We further highlight yet another remarkable potential of coincidence measurements as a probe with better resolution as compared to standard spectroscopic techniques. The finer features show up in both angle resolved and frequency resolved studies.

  19. Chromophore design for large two-photon absorption

    Science.gov (United States)

    Dudley, Christopher

    2014-11-01

    Conjugated oligothiophene chromophores are compared and studied for designing large linear and nonlinear absorption cross-sections. Optical properties of chromophores synthesized by the Naval Research Laboratory are modeled to construct a design factor of merit to predict and understand two-photon absorption (TPA) designs. Computer modeling to optimize parameters to produce photo active chromophores is conducted. Geometry, π-center (electron relay) and the electron donor or acceptor groups attached to the π-centers are considered for importance in TPA. This work could serve equally well as guide for quick back of the envelop research or industrial design verifications as well as an outline for introducing computation methods to students.

  20. New two-photon based nanoscopic modalities and optogenetics

    DEFF Research Database (Denmark)

    Glückstad, Jesper

    -matter interaction on these scales involves the combination of optimal light-sculpting [4] with the use of optimized shapes in micro-robotics structures [5]. Microfabrication processes such as two-photon photo-polymerization offer three-dimensional resolutions for creating custom-designed monolithic microstructures...... that can be equipped with optical trapping handles for convenient mechanical control using only optical forces [6]. These microstructures illustrated above can be effectively handled with simultaneous top- and side-view on our BioPhotonics Workstation to undertake six-degree-of-freedom optical actuation...

  1. Two-photon polymerization of immune cell scaffolds

    DEFF Research Database (Denmark)

    Olsen, Mark Holm

    and easy to use chip integrated migration platform. Free-form constructs with three-dimensional (3D) microporosity were fabricated by two-photon polymerization inside the closed microchannel of an injection molded commercially available polymer chip for analysis of directed cell migration. Acrylate...... also present a poly (ethylene glycol) diacrylate (PEGDA) based strategy to fabricate soft 3D hydrogel scaffolds. Our experiments with the hydrogel confirm we can control the mechanical properties and introduce biochemical cues on the surface that are recognized by fibroblast cells. Finally we present...

  2. The Nelson Model with Less Than Two Photons

    CERN Document Server

    Galtbayar, A; Yajima, K

    2002-01-01

    We study the spectral and scattering theory of the Nelson model for an atom interacting with a photon field in the subspace with less than two photons. For the free electron-photon system, the spectral property of the reduced Hamiltonian in the center of mass coordinates and the large time dynamics are determined. If the electron is under the influence of the nucleus via spatially decaying potentials, we locate the essential spectrum, prove the absence of singular continuous spectrum and the existence of the ground state, and construct wave operators giving the asymptotic dynamics.

  3. Two-photon tomography using on-chip quantum walks

    CERN Document Server

    Titchener, James; Sukhorukov, Andrey

    2016-01-01

    We present a conceptual approach to quantum tomography based on first expanding a quantum state across extra degrees of freedom and then exploiting the introduced sparsity to perform reconstruction. We formulate its application to photonic circuits, and show that measured spatial photon correlations at the output of a specially tailored discrete-continuous quantum-walk can enable full reconstruction of any two-photon spatially entangled and mixed state at the input. This approach does not require any tunable elements, so is well suited for integration with on-chip superconducting photon detectors.

  4. Two Photon Decays of Charmonia from Lattice QCD

    Energy Technology Data Exchange (ETDEWEB)

    Jozef Dudek; Robert Edwards

    2006-07-12

    We make the first calculation in lattice QCD of two-photon decays of mesons. Working in the charmonium sector, using the LSZ reduction to relate a photon to a sum of hadronic vector eigenstates, we compute form-factors in both the space-like and time-like domains for the transitions {eta}{sub c} {yields} {gamma}*{gamma}* and {chi}{sub c0} {yields} {gamma}*{gamma}*. At the on-shell point we find approximate agreement with experimental world-average values.

  5. Quantum teleportation of one- and two-photon superposition states

    Institute of Scientific and Technical Information of China (English)

    李英; 张天才; 张俊香; 谢常德

    2003-01-01

    Quantum teleportation of one- and two-photon superposition states based on EPR entanglement of continuouswave two-mode squeezed state is discussed. The fidelities of teleportation are deduced for two different input quantum states. The dependence of the fidelity on the parameters of EPR entanglement and the gain of the classical channels are shown numerically. Comparing with the teleportation of Fock state and coherent state, it is pointed out that for given EPR entanglement and classical gain, the higher the nonclassicality of the input state, the lower the accessible fidelity of teleportation.

  6. Spectral Features of FM Spectroscopy of Two-Photon Interactions

    Institute of Scientific and Technical Information of China (English)

    夏慧荣; JohnL.Hall

    1994-01-01

    The spectral features of FM two-photon resonant interaction processes have been calculated for five different frequency modulation versions of counter-propagating incident fields. It is found that the proposed new modulation version (case b in the text) provides novel spectral features for a completely canceled absorption and a sharp dispersion shape at the fundamental beat note. Moreover, its absorption feature appears at the second harmonic of the RF modulation frequency generated by the joint modes via six interaction pathways without mutual phase shift. Such features persist even when the effects of the second-order sidebands of the incident fields are taken into account. Application potentials are emphasized.

  7. Inclusive $D*^{+-}$ Production in Two-Photon Collisions at LEP

    CERN Document Server

    Achard, P; Aguilar-Benítez, M; Alcaraz, J; Alemanni, G; Allaby, James V; Aloisio, A; Alviggi, M G; Anderhub, H; Andreev, V P; Anselmo, F; Arefev, A; Azemoon, T; Aziz, T; Bagnaia, P; Bajo, A; Baksay, G; Baksay, L; Baldew, S V; Banerjee, S; Banerjee, Sw; Barczyk, A; Barillère, R; Bartalini, P; Basile, M; Batalova, N; Battiston, R; Bay, A; Becattini, F; Becker, U; Behner, F; Bellucci, L; Berbeco, R; Berdugo, J; Berges, P; Bertucci, B; Betev, B L; Biasini, M; Biglietti, M; Biland, A; Blaising, J J; Blyth, S C; Bobbink, Gerjan J; Böhm, A; Boldizsar, L; Borgia, B; Bottai, S; Bourilkov, D; Bourquin, Maurice; Braccini, S; Branson, J G; Brochu, F; Burger, J D; Burger, W J; Cai, X D; Capell, M; Cara Romeo, G; Carlino, G; Cartacci, A M; Casaus, J; Cavallari, F; Cavallo, N; Cecchi, C; Cerrada, M; Chamizo-Llatas, M; Chang, Y H; Chemarin, M; Chen, A; Chen, G; Chen, G M; Chen, H F; Chen, H S; Chiefari, G; Cifarelli, Luisa; Cindolo, F; Clare, I; Clare, R; Coignet, G; Colino, N; Costantini, S; de la Cruz, B; Cucciarelli, S; van Dalen, J A; De Asmundis, R; Déglon, P L; Debreczeni, J; Degré, A; Deiters, K; Della Volpe, D; Delmeire, E; Denes, P; De Notaristefani, F; De Salvo, A; Diemoz, M; Dierckxsens, M; Dionisi, C; Dittmar, Michael; Doria, A; Dova, M T; Duchesneau, D; Echenard, B; Eline, A; El-Mamouni, H; Engler, A; Eppling, F J; Ewers, A; Extermann, Pierre; Falagán, M A; Falciano, S; Favara, A; Fay, J; Fedin, O; Felcini, Marta; Ferguson, T; Fesefeldt, H S; Fiandrini, E; Field, J H; Filthaut, Frank; 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; Hakobyan, R S; 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; Krenz, 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 A; Lin, C H; Lin, W T; Linde, Frank L; Lista, L; Liu, Z A; Lohmann, W; Longo, E; Lü, Y S; Lübelsmeyer, K; Luci, C; Luminari, L; Lustermann, W; Ma Wen Gan; Malgeri, L; Malinin, A; Maña, C; Mangeol, D J J; 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; Niessen, T; Nisati, A; Nowak, H; Ofierzynski, R A; Organtini, G; Palomares, C; Pandoulas, D; 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; Prokofiev, D O; Prokofev, D; 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; Rosier-Lees, S; Roth, S; Rosenbleck, C; Roux, B; Rubio, Juan Antonio; Ruggiero, G; Rykaczewski, H; Sakharov, A; Saremi, S; Sarkar, S; Salicio, J; Sánchez, E; Sanders, M P; Schäfer, C; Shchegelskii, V; Schmidt-Kärst, S; Schmitz, D; Schopper, Herwig Franz; Schotanus, D J; Schwering, G; Sciacca, C; Servoli, L; Shevchenko, S; Shivarov, N; Shoutko, V; Shumilov, E; Shvorob, A V; Siedenburg, T; Son, D; Spillantini, P; Steuer, M; Stickland, D P; Stoyanov, B; Strässner, A; Sudhakar, K; Sultanov, G G; Sun, L Z; Sushkov, S V; 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, R T; De Walle, M; 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; Wallraff, W; 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; Zilizi, G; Zimmermann, B; Zöller, M

    2002-01-01

    Inclusive D^{*+-} production in two-photon collisions is studied with the L3 detector at LEP, using 683 pb^{-1} of data collected at centre-of-mass energies from 183 to 208 GeV. Differential cross sections are determined as functions of the transverse momentum and pseudorapidity of the D^{*+-} mesons in the kinematic region 1 GeV e^+e^-D^{*+-}X)$ in this kinematical region is measured and the sigma(e^+e^- ---> e^+e^- cc{bar}X) cross section is derived. The measurements are compared with next-to-leading order perturbative QCD calculations.

  8. Two-photon photoassociative spectroscopy of ultracold 88-Sr

    CERN Document Server

    de Escobar, Y N Martinez; Pellegrini, P; Nagel, S B; Traverso, A; Yan, M; Côté, R; Killian, T C

    2008-01-01

    We present results from two-photon photoassociative spectroscopy of the least-bound vibrational level of the X$^1\\Sigma_g^+$ state of the $^{88}$Sr$_2$ dimer. Measurement of the binding energy allows us to determine the s-wave scattering length, $a_{88}=-1.4(6) a_0$. For the intermediate state, we use a bound level on the metastable $^1S_0$-$^3P_1$ potential, which provides large Franck-Condon transition factors and narrow one-photon photoassociative lines that are advantageous for observing quantum-optical effects such as Autler-Townes resonance splittings.

  9. Two-photon photoassociative spectroscopy of ultracold Sr88

    Science.gov (United States)

    Martinez de Escobar, Y. N.; Mickelson, P. G.; Pellegrini, P.; Nagel, S. B.; Traverso, A.; Yan, M.; Côté, R.; Killian, T. C.

    2008-12-01

    We present results from two-photon photoassociative spectroscopy of the least-bound vibrational level of the XΣg+1 state of the Sr288 dimer. Measurement of the binding energy allows us to determine the s -wave scattering length a88=-1.4(6)a0 . For the intermediate state, we use a bound level on the metastable S01-P13 potential, which provides large Franck-Condon transition factors and narrow one-photon photoassociative lines that are advantageous for observing quantum-optical effects such as Autler-Townes resonance splittings.

  10. Conventional and photonic crystal fiber based two-photon fluorescence biosensing

    Science.gov (United States)

    Myaing, Mon Thiri

    Optical fiber probes are widely used in the biomedical field for applications such as optical microscopy, endoscopy, and optical biopsy. Due to their flexibility and small size, optical fibers offer a minimally invasive light interface for imaging and spectroscopic analysis of internal tissue. The development of fluorescent probes for studies of biological processes has increased the importance of developing optical methods for quantitative, in vivo diagnosis. In this dissertation, we discuss the development of a novel two-photon optical fiber fluorescence (TPOFF) probe for real time, in vivo, quantitative fluorescence measurements in biological samples. In order to understand and optimize two-photon excitation through an optical fiber, pulse propagation effects must be considered. We found a simple phenomenological scaling behavior for the energy dependence of the pulse width for negatively pre-chirped pulses propagating in a normally dispersive fiber. As a consequence of this scaling behavior, the dependence of two-photon fluorescence (TPF) on the pulse intensity becomes sub-quadratic. The TPOFF probe employs a scheme where the same single-mode fiber (SMF) is used for both the excitation and collection of TPF. Using this fiber probe, we show quantification of tumor fluorescence both ex vivo and in vivo. In ex vivo measurements of tumors developed from cells expressing the green fluorescence protein (GFP), the TPOFF probe detected fluorescence from tumors with as little as 0.3% GFP cells. These results were similar to flow cytometry analysis of isolated cells from the tumors. The TPOFF measurements of GFP tumors in live, anesthetized mice showed a linear relationship between the measured fluorescence and the percentage of GFP expressing cells. The TPOFF probe was also used in targeted binding experiments of Herceptin antibody and folic acid-dendrimer nanoparticle conjugates. To improve the sensitivity of the TPOFF probe, a double-clad photonic crystal fiber (DCF

  11. Carbon nanodots featuring efficient FRET for two-photon photodynamic cancer therapy with a low fs laser power density.

    Science.gov (United States)

    Wang, Jing; Zhang, Zehui; Zha, Shuai; Zhu, Yinyan; Wu, Peiyi; Ehrenberg, Benjamin; Chen, Ji-Yao

    2014-11-01

    The 5,10,15,20-tetrakis(1-methyl 4-pyridinio) porphyrins (TMPyP), a photosensitizer used for photodynamic therapy of cancers (PDT), were linked to carbon dots (CDots) to form the conjugates of CDot-TMPyP by the electrostatic force. The 415 nm emission band of CDots was well overlapped with the absorption band of TMPyP, so that the Cdots in conjugates can work as donor to transfer the energy to TMPyP moiety by fluorescence resonance energy transfer (FRET) with an FRET efficiency of 45%, determined by the fluorescence lifetime change between the free CDots and conjugated CDots. The two-photon absorption cross section (TPACS) of TMPyP is as low as 110 GM and the TMPyP thus be not suitable for two-photon PDT. Whereas the CDots have high TPACS, and their TPACS are excitation wavelength dependent with the maximum value of 15000 GM at 700 nm. Therefore, the conjugates of CDot-TMPyP were explored for two-photon excitation (TPE) PDT. The two-photon image of CDot-TMPyP in Hela cells was clearly seen under the excitation of a 700 nm femto-second (fs) laser. The singlet oxygen production of CDot-TMPyP was also much higher than that of TMPyP alone under TPE of a 700 nm fs laser. The in vitro PDT killing was further achieved with CDot-TMPyP by TPE of the 700 nm fs laser. Particularly herein the low power density of fs laser from unfocused laser beam was successfully used to carry out the TPE PDT, because of the high TPACS of CDots. These results demonstrate that the CDot-TMPyP conjugates are promising for TPE PDT and needed to investigate further. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. Two-Photon Absorption Spectroscopy of Rubidium with a Dual-Comb Tequnique

    Science.gov (United States)

    Nishiyama, Akiko; Yoshida, Satoru; Hariki, Takuya; Nakajima, Yoshiaki; Minoshima, Kaoru

    2017-06-01

    Dual-comb spectroscopies have great potential for high-resolution molecular and atomic spectroscopies, thanks to the broadband comb spectrum consisting of dense narrow modes. In this study, we apply the dual-comb system to Doppler-free two-photon absorption spectroscopy. The outputs of two frequency combs excite several two-photon transitions of rubidium, and we obtained broadband Doppler-free spectra from dual-comb fluorescence signals. The fluorescence detection scheme circumvents the sensitivity limit which is effectively determined by the dynamic range of photodetectors in absorption-based dual-comb spectroscopies. Our system realized high-sensitive, Doppler-free high-resolution and broadband atomic spectroscopy. A part of observed spectra of 5S_{1/2} - 5D_{5/2} transition is shown in the figure. The hyperfine structures of the F" = 1 - F' = 3,2,1 transitions are fully-resolved and the spectral widths are approximately 5 MHz. The absolute frequency axis is precisely calibrated from comb mode frequencies which were stabilized to a GPS-disciplined clock. This work was supported by JST through the ERATO MINOSHIMA Intelligent Optical Synthesizer Project and Grant-in-Aid for JSPS Fellows (16J02345). A. Nishiyama, S. Yoshida, Y. Nakajima, H. Sasada, K. Nakagawa, A. Onae, K. and Minoshima, Opt. Express 24, 25894 (2016). A. Hipke, S. A. Meek, T. Ideguchi, T.W. Hänsch, and N. Picqué, Phys. Rev. A 90, 011805(R) (2014).

  13. Resonance Two-Photon Ionization of Diarylethene in the Presence of Cyclodextrin

    Directory of Open Access Journals (Sweden)

    Michihiro Hara

    2013-01-01

    Full Text Available The transient absorption of the diarylethene 1,2-bis(2,4-dimethyl-5-phenyl-3-thienyl-3,3,4,4,5,5-hexafluoro-1-cyclopentene (DE was measured using 266 nm laser flash photolysis in the presence of various cyclodextrins (CDs in aqueous solvent. Ionization of DE occurred within the laser flash (5 ns to give the DE radical cation and water-solvated electron. The electron was generated by two-photon excitation through simultaneous irradiation with 266 nm laser light, and the ionization quantum yields of DE in the presence of α-CD, β-CD, m-β-CD, and γ-CD depended on the properties of the DE/CD complexes. These results suggest that useful two-photon ionization occurred for these complexes. The ionization quantum yield of DE was found to increase in aqueous solution after the addition of m-β-CD (50 mM, in which m-β-CD likely controlled the photochromic reactivity of DE.

  14. Two-photon fluorescence and confocal reflected light imaging of thick tissue structures

    Science.gov (United States)

    Kim, Ki H.; So, Peter T. C.; Kochevar, Irene E.; Masters, Barry R.; Gratton, Enrico

    1998-04-01

    The technology of two-photon excitation has opened a window of opportunity for developing non-invasive medical diagnostic tools capable of monitoring thick tissue biochemical states. Using cellular endogenous chromophores, (beta) -nicotinamide- adenine dinucleotide phosphate [NAD(P)H], the cellular metabolic rates in living human skin were determined. Although important functional information can be obtained from the fluorescence spectroscopy of endogenous chromophores, these chromophores are rather poor contrast enhancing agent for mapping cellular morphology. First, most endogenous chromophores are confined to the cellular cytoplasm which prevents the visualization of other cellular organelles. Second, there is significant variability in the distribution and the quantum yield of endogenous chromophores which depends on tissue biochemistry but prevents consistent comparison of cellular morphology. On the other hand, the deep tissue cellular morphology has been imaged with excellent resolution using reflected light confocal microscopy. In reflected light microscopy, the image contrast originates from the index of refraction differences of the cellular structures. The organelle boundaries with significant index differences such as the plasma membrane and the nucleus envelope can be consistently visualized. A combination of morphological and functional information is required for a thorough tissue study. This presentation describes the development of a new microscope which is capable of simultaneously collecting both two-photon fluorescence and confocal reflected light signals. Promising biomedical applications include the non-invasive diagnosis of skin cancer and the study of wound healing.

  15. Multidimensional two-photon imaging and spectroscopy of fresh human bladder biopsies

    Science.gov (United States)

    Cicchi, Riccardo; Crisci, Alfonso; Cosci, Alessandro; Nesi, Gabriella; Giancane, Saverio; Carini, Marco; Pavone, Francesco S.

    2010-02-01

    Two-photon microscopy has been successfully used to image several types of tissues, including skin, muscles, tendons. Nevertheless, its usefulness in imaging bladder tissue has not been investigated yet. In this work we used combined twophoton excited fluorescence, second-harmonic generation microscopy, fluorescence lifetime imaging microscopy, and multispectral two-photon emission detection to investigate different kinds of human ex-vivo fresh biopsies of bladder. Morphological and spectroscopic analyses allowed to characterize both healthy mucosa and carcinoma in-situ samples in a good agreement with common routine histology. Cancer cells showed different morphology with respect to the corresponding healthy cells: they appeared more elongated and with a larger nucleus to cytoplasm ratio. From the spectroscopic point of view, differences between the two tissue types in both spectral emission and fluorescence lifetime distribution were found. Even if further analysis, as well as a more significant statistics on a larger number of samples would be helpful to discriminate between low, mild, and high grade cancer, our method is a promising tool to be used as diagnostic confirmation of histological results, as well to be implemented in a multi-photon endoscope or in a spectroscopic for in in-vivo imaging applications.

  16. Molecular engineering of nanoscale quadrupolar chromophores for two-photon absorption

    Science.gov (United States)

    Porres, Laurent; Mongin, Olivier; Blanchard-Desce, Mireille H.; Ventelon, Lionel; Barzoukas, Marguerite; Moreaux, Laurent; Pons, Thomas; Mertz, Jerome

    2003-02-01

    Our aim has been the design of optimized NLO-phores with very high two-photon absorption (TPA) cross-sections (s2) in the red-NIR region, while maintaining high linear transparency and high fluorescence quantum yield. Our molecular engineering strategy is based on the push-push or pull-pull functionalization of semi-rigid nanoscale conjugated systems. The central building blocks were selected as rigid units that may assist quadrupolar intramolecular charge transfer by acting either as a (weak) donor or acceptor core. Quadrupolar molecules derived either from a phenyl unit, a rigidified biphenyl moiety or a fused bithiophene unit have been considered. Conjugated oligomers made of phenylene-vinylene and/or phenylene-ethynylene units were selected as connecting spacers between the core and the electroactive end groups to ensure effective electronic conjugation while maintaining suitable transparency/fluorescence. The TPA cross-sections were determined by investigating the two-photon-excited fluorescence properties using a Ti:sapphire laser delivering fs pulses. Both the nature of the end groups and of the core moiety play an important role in determining the TPA spectra. In addition, by adjusting the length and nature of the conjugated extensor, both amplification and spectral tuning of TPA cross-sections can be achieved. As a result, push-push fluorophores which demonstrate giant TPA cross-sections (up to 3000 GM) in the visible red, high fluorescence quantum yields and good transparency in the visible range have been obtained.

  17. Optical limiting effect in a two-photon absorption dye doped solid matrix

    Science.gov (United States)

    He, Guang S.; Bhawalkar, Jayant D.; Zhao, Chan F.; Prasad, Paras N.

    1995-10-01

    We recently reported a new lasing dye, trans-4-[p-(N-ethyl-N-hydroxylethylamino)styryl]-N-methylpyridinium tetraphenylborate (ASPT), which has also been shown to possess a strong two-photon absorption (TPA) and subsequent frequency upconversion fluorescence behavior when excited with near infrared laser radiation. Based on the TPA mechanism, a highly efficient optical limiting performance has been demonstrated in a 2 cm long ASPT-doped epoxy rod pumped with 1.06 μm Q-switched laser pulses at 50-250 MW/cm2 intensity levels. The measured nonlinear absorption coefficient reached 6 cm/GW for the tested sample of dopant concentration d0=4×10-3 M/L. The molecular TPA cross section of ASPT in the epoxy matrix is estimated as σ2=2.5×10-18 cm4/GW or σ2'=4.7×10-46 cm4/photon/s, respectively. Two-photon pumped cavity lasing is also observed in an ASPT-doped polymer rod.

  18. Two-photon pumped cavity lasing in novel dye doped bulk matrix rods

    Science.gov (United States)

    He, Guang S.; Zhao, Chan F.; Bhawalkar, Jayant D.; Prasad, Paras N.

    1995-12-01

    Trans-4-[p-(N-ethyl-N-hydroxyethylamino)styryl]-N-methylpyridi that possesses a much greater two-photon absorption cross section and much stronger upconversion fluorescence emission than common organic dyes (such as rhodamine), when excited with near infrared laser radiation. Utilizing ASPT doped bulk polymer rods, two-photon pumped frequency upconverted cavity lasing has been accomplished using a Q-switched Nd:YAG laser as the pump source. The wavelength and pulse duration were ˜600 nm and 3-6 ns, respectively, for the cavity lasing; whereas the corresponding values for pump pulses were 1.06 μm and ˜10 ns, respectively. For a 7 mm long sample rod with a dopant concentration d0=8×10-3 M/L, the conversion efficiency from the absorbed pump energy to the cavity lasing output was ˜3.5% at a pump energy level of 1.3 mJ. The lasing lifetime, in terms of pulse numbers, was more than 4×104 pulses at 2 Hz repetition rate and room temperature.

  19. Two photon fluorescence imaging of lipid membrane domains and potentials using advanced fluorescent probes

    Science.gov (United States)

    Kilin, Vasyl; Darwich, Zeinab; Richert, Ludovic; Didier, Pascal; Klymchenko, Andrey; Mély, Yves

    2013-02-01

    Biomembranes are ordered and dynamic nanoscale structures critical for cell functions. The biological functions of the membranes strongly depend on their physicochemical properties, such as electrostatics, phase state, viscosity, polarity and hydration. These properties are essential for the membrane structure and the proper folding and function of membrane proteins. To monitor these properties, fluorescence techniques and notably, two-photon microscopy appear highly suited due to their exquisite sensitivity and their capability to operate in complex biological systems, such as living cells and tissues. In this context, we have developed multiparametric environment-sensitive fluorescent probes tailored for precise location in the membrane bilayer. We notably developed probes of the 3-hydroxychromone family, characterized by an excited state intramolecular proton transfer reaction, which generates two tautomeric emissive species with well-separated emission bands. As a consequence, the response of these probes to changes in their environment could be monitored through changes in the ratios of the two bands, as well as through changes in the fluorescence lifetimes. Using two-photon ratiometric imaging and FLIM, these probes were used to monitor the surface membrane potential, and were applied to detect apoptotic cells and image membrane domains.

  20. Two-Photon Interactions with Nuclear Breakup in Relativistic Heavy Ion Collisions

    Energy Technology Data Exchange (ETDEWEB)

    Baltz, Anthony J.; Gorbunov, Yuri; R Klein, Spencer; Nystrand, Joakim

    2010-07-07

    Highly charged relativistic heavy ions have high cross-sections for two-photon interactions. The photon flux is high enough that two-photon interactions may be accompanied by additional photonuclear interactions. Except for the shared impact parameter, these interactions are independent. Additional interactions like mutual Coulomb excitation are of experimental interest, since the neutrons from the nuclear dissociation provide a simple, relatively unbiased trigger. We calculate the cross sections, rapidity, mass and transverse momentum (p{sub T}) distributions for exclusive {gamma}{gamma} production of mesons and lepton pairs, and for {gamma}{gamma} reactions accompanied by mutual Coulomb dissociation. The cross-sections for {gamma}{gamma} interactions accompanied by multiple neutron emission (XnXn) and single neutron emission (1n1n) are about 1/10 and 1/100 of that for the unaccompanied {gamma}{gamma} interactions. We discuss the accuracy with which these cross-sections may be calculated. The typical p{sub T} of {gamma}{gamma} final states is several times smaller than for comparable coherent photonuclear interactions, so p{sub T} may be an effective tool for separating the two classes of interactions.

  1. Polarization-Sensitive Two-Photon Microscopy Study of the Organization of Liquid-Crystalline DNA

    Science.gov (United States)

    Mojzisova, Halina; Olesiak, Joanna; Zielinski, Marcin; Matczyszyn, Katarzyna; Chauvat, Dominique; Zyss, Joseph

    2009-01-01

    Abstract Highly concentrated DNA solutions exhibit self-ordering properties such as the generation of liquid-crystalline phases. Such organized domains may play an important role in the global chromatin topology but can also be used as a simple model for the study of more complex 3D DNA structures. In this work, using polarized two-photon fluorescence microscopy, we report on the orientation of DNA molecules in liquid-crystalline phases. For this purpose, we analyze the signal emitted by fluorophores that are noncovalently bound to DNA strands. In nonlinear processes, excitation occurs exclusively in the focal volume, which offers advantages such as the reduction of photobleaching of out-of-focus molecules and intrinsic 3D sectioning capability. Propidium iodide and Hoechst, two fluorophores with different DNA binding modes, have been considered. Polarimetric measurements show that the dyes follow the alignment with respect to the DNA strands and allow the determination of the angles between the emission dipoles and the longitudinal axis of the DNA double strand. These results provide a useful starting point toward the application of two-photon polarimetry techniques to determine the local orientation of condensed DNA in physiological conditions. PMID:19843467

  2. Two-photon absorption and transient photothermal imaging of pigments in tissues

    Science.gov (United States)

    Ye, Tong; Fu, Dan; Matthews, Thomas E.; Hong, Lian; Simon, John D.; Warren, Warren S.

    2008-02-01

    As a main pigment in skin tissues, melanin plays an important role in photo-protecting skin from UV radiation. However, melanogenesis may be altered due to disease or environmental factors; for example, sun exposure may cause damage and mutation of melanocytes and induce melanoma. Imaging pigmentation changes may provide invaluable information to catch the malignant transformation in its early stage and in turn improve the prognosis of patients. We have demonstrated previously that transmission mode, two-photon, one- or two-color absorption microscopy could provide remarkable contrast in imaging melanin in skin. In this report we demonstrate significantly improved sensitivity, so that we are now able to image in epi-mode (or back reflection) in two-photon absorption. This improvement makes possible for us to characterize the different types of pigmentation on the skin in vivo at virtually any location. Another finding is that we can also image transient photothermal dynamics due to the light absorption of melanin. By carefully choosing excitation and probe wavelengths, we might be able to image melanin in different structures under different micro-environments in skin, which could provide useful photochemical and photophysical insights in understanding how pigments are involved in photoprotection and photodamage of cells.

  3. Phase- and Polarization-Controlled Two-Photon Rabi Oscillation of the Biexciton State in a Semiconductor Quantum Dot

    Directory of Open Access Journals (Sweden)

    Erlin Sun

    2014-01-01

    Full Text Available Under a degenerate two-photon resonant excitation, the Rabi oscillation of the four-level biexciton system in a semiconductor quantum dot is theoretically investigated. The influence of the laser phases on the state manipulation is modeled and numerically calculated. Due to the interference between different excitation paths, the laser phase plays an important role and can be utilized as an alternate control knob to coherently manipulate the biexciton state. The phase control can be facilely implemented by changing the light polarization via a quarter-wave plate.

  4. Dermal factors influencing measurement of skin autofluorescence

    NARCIS (Netherlands)

    Noordzij, Margaretha J.; Lefrandt, Johan; Graaff, Reindert; Smit, Andries J.

    Background: Skin autofluorescence (SAF) is a noninvasive marker of accumulation of advanced glycation end products. It predicts cardiovascular complications and mortality in diabetes and renal failure. We assessed the influence of potential common confounders in SAF measurement, by determining the

  5. Simple noninvasive measurement of skin autofluorescence.

    Science.gov (United States)

    Meerwaldt, Robbert; Links, Thera; Graaff, Reindert; Thorpe, Suzannne R; Baynes, John W; Hartog, Jasper; Gans, Reinold; Smit, Andries

    2005-06-01

    Accumulation of advanced glycation end products (AGEs) is thought to play a role in the pathogenesis of chronic complications of diabetes mellitus and renal failure. Several studies indicate that AGE accumulation in tissue may reflect the cumulative effect of hyperglycemia and oxidative stress over many years. Simple quantitation of AGE accumulation in tissue could provide a tool for assessing the risk of long-term complications. Because several AGEs exhibit autofluorescence, we developed a noninvasive autofluorescence reader (AFR). Skin autofluorescence measured with the AFR correlates with collagen-linked fluorescence and specific skin AGE levels from skin biopsy samples. Furthermore, skin autofluorescence correlates with long-term glycemic control and renal function, and preliminary results show correlations with the presence of long-term complications in diabetes. The AFR may be useful as a clinical tool for rapid assessment of risk for AGE-related long-term complications in diabetes and in other conditions associated with AGE accumulation.

  6. Autofluorescent parameters of the skin changes in accordance of age and insolation

    Directory of Open Access Journals (Sweden)

    E. V. Krylova

    2015-01-01

    Full Text Available There are several key factors, such as age and sun exposure, the effects of which leads to a change in the properties of the skin, detected by the method of autofluorescent dermoscopy. In this paper we used the principle of multispectral imaging of the skin in the light of autofluorescence at excitation wavelength of about 400 nm. It was found that, the background skin glow, the coefficient of heterogeneity and the integral index of autofluorescence significantly increases in a low insolated areas. The most pronounced increase of these parameters observed in the group of women older than 45 years. We can assume that this is due to changes in the properties of collagen and accumulation of advanced glycation end products, and the uneven distribution of absorbent skin chromophores. Insolation accelerates the development of age-related changes of skin AF. According to the literature, in people aged 50–70 years increased NADH (external NADH oxidase and AGEs compared with age 20–40 years. The differences in the autofluorescence of the skin between high and low insolated sites saved in all age groups. Autofluorescence age-related features described above should be considered for diagnostic and therapeutic measures. 

  7. Biological imaging without autofluorescence in the second near-infrared region

    Institute of Scientific and Technical Information of China (English)

    Shuo Diao[1; Guosong Hong[1; Alexander L. Antaris[1; Jeffrey L. Blackbum[2; Kai Cheng[3; Zhen Cheng[3; Hongjie Dai[1

    2015-01-01

    Fluorescence imaging is capable of acquiring anatomical and functional infor- mation with high spatial and temporal resolution. This imaging technique has been indispensable in biological research and disease detection/diagnosis. Imaging in the visible and to a lesser degree, in the near-infrared (NIR) regions below 900 nm, suffers from autofluorescence arising from endogenous fluorescent molecules in biological tissues. This autofluorescence interferes with fluorescent molecules of interest, causing a high background and low detection sensitivity. Here, we report that fluorescence imaging in the 1,500-1,700-nm region (termed "NIR-IIb") under 808-nm excitation results in nearly zero tissue autofluorescence, allowing for background-free imaging of fluorescent species in otherwise notoriously autofluorescent biological tissues, including liver. Imaging of the intrinsic fluorescence of individual fluorophores, such as a single carbon nanotube, can be readily achieved with high sensitivity and without autofluorescence background in mouse liver within the 1,500-1,700-nm wavelength region.

  8. Two-Photon Holographic Stimulation of ReaChR

    Science.gov (United States)

    Chaigneau, Emmanuelle; Ronzitti, Emiliano; Gajowa, Marta A.; Soler-Llavina, Gilberto J.; Tanese, Dimitrii; Brureau, Anthony Y. B.; Papagiakoumou, Eirini; Zeng, Hongkui; Emiliani, Valentina

    2016-01-01

    Optogenetics provides a unique approach to remotely manipulate brain activity with light. Reaching the degree of spatiotemporal control necessary to dissect the role of individual cells in neuronal networks, some of which reside deep in the brain, requires joint progress in opsin engineering and light sculpting methods. Here we investigate for the first time two-photon stimulation of the red-shifted opsin ReaChR. We use two-photon (2P) holographic illumination to control the activation of individually chosen neurons expressing ReaChR in acute brain slices. We demonstrated reliable action potential generation in ReaChR-expressing neurons and studied holographic 2P-evoked spiking performances depending on illumination power and pulse width using an amplified laser and a standard femtosecond Ti:Sapphire oscillator laser. These findings provide detailed knowledge of ReaChR's behavior under 2P illumination paving the way for achieving in depth remote control of multiple cells with high spatiotemporal resolution deep within scattering tissue. PMID:27803649

  9. Inclusive D*(+/-) production in two photon collisions at LEP

    CERN Document Server

    Prokofiev, Denis Olegovich

    2001-01-01

    In this thesis I present my results on the measurement of the open charm production in two-photon collision events done with the L3 detector at Large Electron Positron machine (LEP). The data sample was collected from 1997 through 2000 at center-of-mass energies ranging from 183 GeV to 209 GeV, corresponding to a total integrated luminosity of 683.4pb −1. The open charm production in two-photon collision events extrapolated to the full phase space is estimated to be: s&parl0;e+e-&rarrr;e +e-cc&d1;X&parr0;=9 23±69±109±222pb. The differential cross sections d s /dpT(D*±) and d s /d:η(D*±): are also measured as functions of transverse momentum pT(D*±) and the absolute value of pseudorapidity :η(D*±):, respectively. A fit to the data estimating the relative contributions of Direct and Resolved open charm production mechanisms is performed, giving (28.7 ± 5.6)% and (71.3 ± 8.8)%, respectively. Using those relative fractions, the Direct and Resolved process cross sections yield: s&p...

  10. High-order dispersion effects in two-photon interference

    Science.gov (United States)

    Mazzotta, Zeudi; Cialdi, Simone; Cipriani, Daniele; Olivares, Stefano; Paris, Matteo G. A.

    2016-12-01

    Two-photon interference and Hong-Ou-Mandel (HOM) effect are relevant tools for quantum metrology and quantum information processing. In optical coherence tomography, the HOM effect is exploited to achieve high-resolution measurements with the width of the HOM dip being the main parameter. On the other hand, applications like dense coding require high-visibility performance. Here we address high-order dispersion effects in two-photon interference and study, theoretically and experimentally, the dependence of the visibility and the width of the HOM dip on both the pump spectrum and the downconverted photon spectrum. In particular, a spatial light modulator is exploited to experimentally introduce and manipulate a custom phase function to simulate the high-order dispersion effects. Overall, we show that it is possible to effectively introduce high-order dispersion effects on the propagation of photons and also to compensate for such effect. Our results clarify the role of the different dispersion phenomena and pave the way for optimization procedures in quantum technological applications involving PDC photons and optical fibers.

  11. Theory of Two-Photon Absorptions in Graphene Fragments

    Science.gov (United States)

    Aryanpour, K.; Shukla, A.; Mazumdar, S.; Sandhu, A.; Roberts, A.

    2012-02-01

    Electron-electron correlations in graphene is currently an active field of research [1-3]. The carbon atoms in graphene have the same sp^2 hybridization as in strongly correlated π-conjugated polymer systems. The low energy behavior in graphene however appears to be reasonably described within the one-electron Dirac massless fermions model. Historically, the occurrence of the lowest two-photon state below the optical one-photon state provided the strongest proof for strong electron correlations in linear polyenes [4]. We systematically study the Coulomb interaction effects on the ground state and nonlinear absorptions in graphene fragments as a function of system size, beginning from the smallest stable fragment coronene. We report high order calculations of one- vs two-photon spin singlet and triplet states, in coronene, hexabenzocoronene and other molecular fragments that clearly indicate the strong role of electron-electron interactions. We will discuss the implications of our work on molecular systems for the thermodynamic limit of graphene. [4pt] [1] Siegel David A.; et al., PNAS, v108, 28, 11365-11369 (2011)[0pt] [2] Gr"onqvist J. H.; et al., arXiv: 1107.5653v1[0pt] [3] Uchoa B.; et al., arXiv: 1109.1577v1[0pt] [4] Ramasesha S.; et al., J. Chem. Phys. 80, 3278 (1984)

  12. Nonresonant two-photon transitions in length and velocity gauges

    Science.gov (United States)

    Jentschura, U. D.

    2016-08-01

    We reexamine the invariance of two-photon transition matrix elements and corresponding two-photon Rabi frequencies under the "gauge" transformation from the length to the velocity gauge. It is shown that gauge invariance, in the most general sense, only holds at exact resonance, for both one-color as well as two-color absorption. The arguments leading to this conclusion are supported by analytic calculations which express the matrix elements in terms of hypergeometric functions, and ramified by a "master identity" which is fulfilled by off-diagonal matrix elements of the Schrödinger propagator under the transformation from the velocity to the length gauge. The study of the gauge dependence of atomic processes highlights subtle connections between the concept of asymptotic states, the gauge transformation of the wave function, and infinitesimal damping parameters for perturbations and interaction Hamiltonians that switch off the terms in the infinite past and future [of the form exp(-ɛ |t |)] . We include a pertinent discussion.

  13. A [111]-Cut Si Hemisphere Two-Photon Response Photodetector

    Institute of Scientific and Technical Information of China (English)

    LIU Xiu-Huan; CHEN Zhan-Guo; JIA Gang; WANG Hai-Yan; GAO Yan-Jun; LI Yi1

    2011-01-01

    Properties of two-photon response in a [lll]-cut nearly-intrinsic Si hemisphere photodetector are studied. The measured photocurrent of the photodetector responding to the 1.32μm continuous wave laser shows a quadratic dependence on the coupled optical power and is saturated with the bias voitage. Also, the photocurrent is independent of polarization. Such properties are in good agreement with the theory of two-photon absorption. The isotropic photocurrent generated from the [lll]-cut Si hemisphere is compared to the anisotropic one induced in the [110]-cut Si sample and the ratio of Xxxxx /Xxxyy for silicon performing at 1.32μm is calculated to be 2.4 via the fitted function of the anisotropic photocurrent from the [110]-cut sample.%Properties of two-photon response in a [111]-cut nearly-intrinsic Si hemisphere photodetector are studied.The measured photocurrent of the photodetector responding to the 1.32 μm continuous wave laser shows a quadratic dependence on the coupled optical power and is saturated with the bias voltage.Also,the photocurrent is independent of polarization.Such properties are in good agreement with the theory of two-photon absorption.The isotropic photocurrent generated from the [111]-cut Si hemisphere is compared to the anisotropic one induced in the [110]-cut Si sample and the ratio of Xxxxx /Xxxyy for silicon performing at 1.32μm is calculated to be 2.4via the fitted function of the anisotropic photocurrent from the [110]-cut sample.Silicon materials have a variety of applications in microelectronics and silicon optoelectronics and are still attractive to relevant researchers.Commercial Si photodetectors are largely designed based on singlephoton absorption (SPA).However,nonlinear characteristics have been exhibited in silicon devices.Specifically,two-photon absorption (TPA) has attracted much attention in such devices of Si p-n and p-i-n photodiodes,Si waveguides and Si avalanche diodes,etc.for the autocorrelation measurements of

  14. Measurement of Ultra-Short Single-Photon Pulse Duration with Two-Photon Interference

    Institute of Scientific and Technical Information of China (English)

    LV Fan; SUN Fang-Wen; ZOU Chang-Ling; HAN Zheng-Fu; GUO Guang-Can

    2011-01-01

    We proposed a protocol of measuring the duration of ultra-short single-photon pulse with two-photon interference.The pulse duration can be obtained from the width of the visibility of two-photon Hong-Ou-Mandel interference or the indistinguishability of the two photons. Moreover, the shape of a single-photon pulse can be measured with ultra-short single-photon pulses through the two-photon interference.%@@ We proposed a protocol of measuring the duration of ultra-short single-photon pulse with two-photon interference.The pulse duration can be obtained from the width of the visibility of two-photon Hong-Ou-Mandel interference or the indistinguishability of the two photons.Moreover, the shape of a single-photon pulse can be measured with ultra-short single-photon pulses through the two-photon interference.

  15. Laser-induced dental caries and plaque diagnosis on patients by sensitive autofluorescence spectroscopy and time-gated video imaging: preliminary studies

    Science.gov (United States)

    Koenig, Karsten; Schneckenburger, Herbert

    1994-09-01

    The laser-induced in vivo autofluorescence of human teeth was investigated by means of time- resolved/time-gated fluorescence techniques. The aim of these studies was non-contact caries and plaque detection. Carious lesions and dental plaque fluoresce in the red spectral region. This autofluorescence seems to be based on porphyrin-producing bacteria. We report on preliminary studies on patients using a novel method of autofluorescence imaging. A special device was constructed for time-gated video imaging. Nanosecond laser pulses for fluorescence excitation were provided by a frequency-doubled, Q-switched Nd:YAG laser. Autofluorescence was detected in an appropriate nanosecond time window using a video camera with a time-gated image intensifier (minimal time gate: 5 ns). Laser-induced autofluorescence based on porphyrin-producing bacteria seems to be an appropriate tool for detecting dental lesions and for creating `caries-images' and `dental plaque' images.

  16. Brominated 7-hydroxycoumarin-4-ylmethyls: Photolabile protecting groups with biologically useful cross-sections for two photon photolysis

    Science.gov (United States)

    Furuta, Toshiaki; Wang, Samuel S.-H.; Dantzker, Jami L.; Dore, Timothy M.; Bybee, Wendy J.; Callaway, Edward M.; Denk, Winfried; Tsien, Roger Y.

    1999-01-01

    Photochemical release (uncaging) of bioactive messengers with three-dimensional spatial resolution in light-scattering media would be greatly facilitated if the photolysis could be powered by pairs of IR photons rather than the customary single UV photons. The quadratic dependence on light intensity would confine the photolysis to the focus point of the laser, and the longer wavelengths would be much less affected by scattering. However, previous caged messengers have had very small cross sections for two-photon excitation in the IR region. We now show that brominated 7-hydroxycoumarin-4-ylmethyl esters and carbamates efficiently release carboxylates and amines on photolysis, with one- and two-photon cross sections up to one or two orders of magnitude better than previously available. These advantages are demonstrated on neurons in brain slices from rat cortex and hippocampus excited by glutamate uncaged from N-(6-bromo-7-hydroxycoumarin-4-ylmethoxycarbonyl)-l-glutamate (Bhc-glu). Conventional UV photolysis of Bhc-glu requires less than one-fifth the intensities needed by one of the best previous caged glutamates, γ-(α-carboxy-2-nitrobenzyl)-l-glutamate (CNB-glu). Two-photon photolysis with raster-scanned femtosecond IR pulses gives the first three-dimensionally resolved maps of the glutamate sensitivity of neurons in intact slices. Bhc-glu and analogs should allow more efficient and three-dimensionally localized uncaging and photocleavage, not only in cell biology and neurobiology but also in many technological applications. PMID:9990000

  17. Folate-receptor-mediated delivery of InP quantum dots for bioimaging using confocal and two-photon microscopy.

    Science.gov (United States)

    Bharali, Dhruba J; Lucey, Derrick W; Jayakumar, Harishankar; Pudavar, Haridas E; Prasad, Paras N

    2005-08-17

    A novel method for the synthesis of highly monodispersed hydrophillic InP-ZnS nanocrystals and their use as luminescence probes for live cell imaging is reported. Hydrophobic InP-ZnS nanocrystals are prepared by a new method that yields high-quality, luminescent core-shell nanocrystals within 6-8 h of total reaction time. Then by carefully manipulating the surface of these passivated nanocrystals, aqueous dispersions of folate-conjugated nanocrystals (folate-QDs) with high photostability are prepared. By use of confocal microscopy, we demonstrate the receptor-mediated delivery of folic acid conjugated quantum dots into folate-receptor-positive cell lines such as KB cells. These folate-QDs tend to accumulate in multi-vescicular bodies of KB cells after 6 h of incubation. Receptor-mediated delivery was confirmed by comparison with the uptake of these particles in folate-receptor-negative cell lines such as A549. Efficient two-photon excitation of these particles and two-photon imaging using these particles are also demonstrated. The use of these InP-ZnS nanoparticles and their efficient two-photon excitation can be potentially useful for deep tissue imaging for future in vivo studies.

  18. Ab initio study of the one- and two-photon circular dichroism of R-(+)-3-methyl-cyclopentanone

    Science.gov (United States)

    Rizzo, Antonio; Lin, Na; Ruud, Kenneth

    2008-04-01

    One- and two-photon circular dichroism spectra of R-(+)-3-methyl-cyclopentanone, a system that has been the subject of recent experimental studies of (2+1) resonance-enhanced multiphoton ionization circular dichroism, have been calculated with an origin-invariant density functional theory approximation in the region of the lowest electronic excited states, both for the gas phase and for a selection of solvents. A polarizable continuum model is used in the calculations performed on the solvated system. Two low-lying conformers are analyzed, and a comparison of the intensities and characteristic features is made with the corresponding two-photon absorption for each species, also for the Boltzmann-averaged spectra. The effect of the choice of geometry, basis set, and exchange-correlation functional is carefully analyzed. It is found that a density functional theory approach using the Coulomb attenuating method variant of Becke's three-parameter exchange and the Lee-Yang-Parr correlation functionals with correlation-consistent basis sets of double-zeta quality can reproduce the experimental electronic circular dichroism spectra very well. The features appearing in experiment are characterized in terms of molecular excitations, and the differences in the response of each state in the one- and two-photon processes are highlighted.

  19. Quantitative Imaging of Molecular Order in Lipid Membranes Using Two-Photon Fluorescence Polarimetry

    Science.gov (United States)

    Gasecka, Alicja; Han, Tsai-Jung; Favard, Cyril; Cho, Bong Rae; Brasselet, Sophie

    2009-01-01

    Abstract We present a polarimetric two-photon microscopy technique to quantitatively image the local static molecular orientational behavior in lipid and cell membranes. This approach, based on a tunable excitation polarization state complemented by a polarized readout, is easily implementable and does not require hypotheses on the molecular angular distribution such as its mean orientation, which is a main limitation in traditional fluorescence anisotropy measurements. The method is applied to the investigation of the molecular angular distribution in giant unilamellar vesicles formed by liquid-ordered and liquid-disordered micro-domains, and in COS-7 cell membranes. The highest order contrast between ordered and disordered domains is obtained for dyes locating within the membrane acyl chains. PMID:19917241

  20. Two Photon Induced Lasing in 1550 nm Quantum Dash Optical Gain Media

    DEFF Research Database (Denmark)

    Capua, Amir; Saal, Abigael; Reithmaier, Johann Peter

    2011-01-01

    We report on a unique lasing mechanism observed in quantum dash Gain media. While the gain media is electrically pumped below lasing threshold, a strong optical pulse excites carriers by two photon absorption into high energy states of the quantum dashes and wetting layer. Fast inter band carrier...... by the XFROG scheme is performed. We show the lasing mechanism to be governed mainly by the wetting layer dynamics and extract a direct measurement of the carrier-carrier scattering time constant....... relaxation and capture processes into the ground states of the quantum dashes result in increased gain followed by lasing at the gain peak irrespective of the stimulating pulse wavelength. The temporal response of the lasing line is examined on a 40 GHz scope and full characterization of the pulse...

  1. PHOTOCHROMISM AND LUMINESCENCE OF DOPANT CHROMOPHORES THROUGH TWO-PHOTON IONIZATION IN POLYMER FILMS

    Institute of Scientific and Technical Information of China (English)

    Masahide Yamamoto; Hideo Ohkita; Shinzaburo Ito

    2001-01-01

    Two-photon ionization and recombination processes of an aromatic chromophore doped in polymer films were studied and the features of these processes were discussed in relation to photofunctional polymers. An aromatic molecule having low ionization potential, e.g., N,N,N',N'-tetramethyl-p-phenylene diamine doped in poly(methyl methacrylate)(PMMA) film was easily photoionized by intense laser light excitation, giving a colored radical cation (photochromism) and a trapped electron in PMMA matrix. As a reversed process, the radical cation recombined with the trapped electron, showing discoloration and emitting luminescence, either isothermal luminescence (ITL), or thermoluminescence (TL). In this report,ITL and TL through the charge recombination process were studied and the luminescence was suggested as a mean of the read-out of photorecording.

  2. Numerical approaches for predicting two-photon absorption induced single-event effects in semiconductors

    Science.gov (United States)

    Hales, Joel M.; Khachatrian, Ani; Roche, Nicolas J.; Buchner, Stephen; Warner, Jeffrey; McMorrow, Dale

    2016-05-01

    Two numerical approaches for determining the charge generated in semiconductors via two-photon absorption (2PA) under conditions relevant for laser-based single-event effects (SEE) experiments are presented. The first approach uses a simple analytical expression incorporating a small number of experimental/material parameters while the second approach employs a comprehensive beam propagation method that accounts for all the complex nonlinear optical (NLO) interactions present. The impact of the excitation conditions, device geometry, and specific NLO interactions on the resulting collected charge in silicon devices is also discussed. These approaches can provide value to the radiation-effects community by predicting the impacts that varying experimental parameters will have on 2PA SEE measurements.

  3. Development and application of biological techniques to two-photon photodynamic therapy

    Science.gov (United States)

    Khurana, Mamta; Karotki, Aliaksandr; Moriyama, Eduardo H.; Akens, Margarete K.; Wilson, Brian C.

    2007-06-01

    Two-photon (2-γ) photodynamic therapy (PDT) as opposed to "standard" one-photon (1-γ) PDT with Visudyne has recently been suggested as a targeted treatment alternative for wet-form age-related macular degeneration (AMD) and other neovascular diseases. AMD is a major cause of severe vision loss in the older population. It occurs due to growth of new leaky blood vessels (neovasculature) from the choriocapillaris, which results in destruction of photoreceptors in the fovea and loss of central vision. Damage outside the diseased region is always a concern, due to photosensitizer accumulation and its 1-γ excitation. Highly targeted 2-γ excitation, due to its non-linear intensity dependence, intrinsically avoids out-of-focus damage to healthy tissues and so could be valuable for wet-AMD. We have previously developed a quantitative approach for comparing the 2-γ efficacy of photosensitizers in vitro. In this study, we report further the development of ex vivo and in vivo techniques. A mouse mesenteric vessel has been investigated as the ex vivo model of neovasculature. For the in vivo studies, we have explored a mouse dorsal skin-fold window chamber model. Two-photon PDT is delivered using tightly focused ~300 fs laser pulses from a Ti:sapphire laser operating at 850 nm with 90 MHz pulse repetition rate. Confocal microscopy coupled to the laser was used to visualize the vessel's/cell's response before, during and after the treatment. We are able to demonstrate quantitative biological techniques to evaluate efficacy of 2-γ PDT photosensitizers in vivo.

  4. Electromagnetically induced absorption and transparency in an optical-rf two-photon coupling configuration

    Energy Technology Data Exchange (ETDEWEB)

    Fu Guangsheng [College of Physical Science and Technology, Hebei University, Baoding 071002 (China); Li Xiaoli [College of Physical Science and Technology, Hebei University, Baoding 071002 (China)], E-mail: xiaolixiaoli001@yahoo.com.cn; Zhuang Zhonghong; Zhang Lianshui; Yang Lijun; Li Xiaowei; Han Li [College of Physical Science and Technology, Hebei University, Baoding 071002 (China); Manson, Neil B.; Wei Changjiang [Laser Physics Center, Research School of Physical Sciences and Engineering, Australian Nation University, Canberra, ACT 0200 (Australia)

    2008-01-07

    We study electromagnetically induced absorption (EIA) and transparency (EIT) in an optical-rf two-photon coupling configuration. It is shown that the interference effect due to interacting dark resonances results in an EIA for a resonant two-photon coupling and this EIA is observed to evolve into an EIT when there is a detuning in the two-photon coupling.

  5. NIR-to-NIR Two-Photon Scanning Laser Microscopy Imaging of Single Nanoparticles Doped by Yb(III) Complexes.

    Science.gov (United States)

    Bourdolle, Adrien; D'Aléo, Anthony; Philippot, Cécile; Baldeck, Patrice L; Guyot, Yannick; Dubois, Fabien; Ibanez, Alain; Andraud, Chantal; Brasselet, Sophie; Maury, Olivier

    2016-01-04

    The photophysical and nonlinear optical properties of water-soluble chromophore-functionalised tris-dipicolinate complexes [LnL3](3-) (Ln=Yb and Nd) are thoroughly studied, revealing that only the Yb(III) luminescence can be sensitized by a two-photon excitation process. The stability of the complex in water is strongly enhanced by embedding in dispersible organosilicate nanoparticles (NPs). Finally, the spectroscopic properties of [NBu4]3 [YbL3] are studied in solution and in the solid state. The high brightness of the NPs allows imaging them as single objects using a modified two-photon microscopy setup in a NIR-to-NIR configuration.

  6. Coverage-dependent two-photon photoexcitation at the H2O/TiO2 interface

    Science.gov (United States)

    Payne, D. T.; Zhang, Y.; Pang, C. L.; Fielding, H. H.; Thornton, G.

    2016-10-01

    Excited electrons and holes are crucial for redox reactions on metal oxide surfaces. However, precise details of this charge transfer process are not known. We report two-photon photoemission (hν = 3.23 eV) measurements of rutile TiO2(110) as a function of exposure to water below room temperature. The two-photon resonance associated with bridging hydroxyls is enhanced following water exposure, reaching a maximum at a nominal coverage of one monolayer. Higher coverages attenuate the observed resonance. Ultraviolet photoemission spectroscopy (hν = 21.22 eV) of the initial, band gap states shows little change up to one monolayer water coverage. It is likely that the enhancement arises from dissociation within the adsorbed water monolayer, although other mechanisms cannot be excluded.

  7. Two-photon fluorescence and second-harmonic generation imaging of collagen in human tissue based on multiphoton microscopy.

    Science.gov (United States)

    Jiang, Xingshan; Zhong, Jiazhao; Liu, Yuchun; Yu, Haibo; Zhuo, Shuangmu; Chen, Jianxin

    2011-01-01

    Multiphoton microscopic imaging of collagen plays an important role in noninvasive diagnoses of human tissue. In this study, two-photon fluorescence and second-harmonic generation (SHG) imaging of collagen in human skin dermis and submucosa of colon and stomach tissues were investigated based on multiphoton microscopy (MPM). Our results show that multiphoton microscopic image of collagen bundles exhibits apparently different pattern in human tissues. The collagen bundles can simultaneously reveal its SHG and two-photon excited fluorescence images in the submucosa of colon and stomach, whereas it solely emit SHG signal in skin dermis. The intensity spectral information from tissues further demonstrated the above results. This indicates that collagen bundles have completely different space arrangement in these tissues. Our experimental results bring more detailed information of collagen for the application of MPM in human noninvasive imaging. Copyright © 2011 Wiley Periodicals, Inc.

  8. Cellular imaging of deep organ using two-photon Bessel light-sheet nonlinear structured illumination microscopy

    Science.gov (United States)

    Zhao, Ming; Zhang, Han; Li, Yu; Ashok, Amit; Liang, Rongguang; Zhou, Weibin; Peng, Leilei

    2014-01-01

    In vivo fluorescent cellular imaging of deep internal organs is highly challenging, because the excitation needs to penetrate through strong scattering tissue and the emission signal is degraded significantly by photon diffusion induced by tissue-scattering. We report that by combining two-photon Bessel light-sheet microscopy with nonlinear structured illumination microscopy (SIM), live samples up to 600 microns wide can be imaged by light-sheet microscopy with 500 microns penetration depth, and diffused background in deep tissue light-sheet imaging can be reduced to obtain clear images at cellular resolution in depth beyond 200 microns. We demonstrate in vivo two-color imaging of pronephric glomeruli and vasculature of zebrafish kidney, whose cellular structures located at the center of the fish body are revealed in high clarity by two-color two-photon Bessel light-sheet SIM. PMID:24876996

  9. Novel Bis-β-diketone-type Ligand and Its Copper and Zinc Complexes for Two-photon Biological Imaging

    Institute of Scientific and Technical Information of China (English)

    ZHOU Shuang-sheng; XUE Xuan; WEI Dong; JIANG Bo; WANG Jia-feng; LU Cheng-hua

    2012-01-01

    A curcumin derivative ligand,1,7-bis(3-methoxyl-4-oxyethylacetate)phenyl-1,6-heptadiene-3,5-diketone (diethyl acetatecurcumin,abbreviated as HL),and its Cu(Ⅱ) and Zn(Ⅱ) complexes have been synthesized and characterized by elemental analyses,infrared(IR),1H NMR and molar conductivity.The experimental results show that the resulting complexes bear strong two-photon excited fluorescence(TPEF) in N,N-dimethyformamide solvent,which has been proven to be potentially useful for two-photon microscopy imaging in living cells.In addition,cytotoxicity tests show that the low-micromolar concentrations of metal-ligand complex(ML2) did not cause significant reduction in cell viability over a pcriod of,at least,24 h and should be safe for further biological studies.

  10. Cellular imaging of deep organ using two-photon Bessel light-sheet nonlinear structured illumination microscopy.

    Science.gov (United States)

    Zhao, Ming; Zhang, Han; Li, Yu; Ashok, Amit; Liang, Rongguang; Zhou, Weibin; Peng, Leilei

    2014-05-01

    In vivo fluorescent cellular imaging of deep internal organs is highly challenging, because the excitation needs to penetrate through strong scattering tissue and the emission signal is degraded significantly by photon diffusion induced by tissue-scattering. We report that by combining two-photon Bessel light-sheet microscopy with nonlinear structured illumination microscopy (SIM), live samples up to 600 microns wide can be imaged by light-sheet microscopy with 500 microns penetration depth, and diffused background in deep tissue light-sheet imaging can be reduced to obtain clear images at cellular resolution in depth beyond 200 microns. We demonstrate in vivo two-color imaging of pronephric glomeruli and vasculature of zebrafish kidney, whose cellular structures located at the center of the fish body are revealed in high clarity by two-color two-photon Bessel light-sheet SIM.

  11. Characterizing the intracellular distribution of metabolites in intact Chlamydia-infected cells by Raman and two-photon microscopy.

    Science.gov (United States)

    Szaszák, Márta; Chang, Jiun Chiun; Leng, Weinan; Rupp, Jan; Ojcius, David M; Kelley, Anne Myers

    2013-06-01

    Chlamydia species are obligate intracellular pathogens that proliferate only within infected cells. Currently, there are no known techniques or systems that can probe the spatial distribution of metabolites of interest within intact Chlamydia-infected cells. Here we investigate the ability of Raman microscopy to probe the chemical composition of different compartments (nucleus, inclusion, and cytoplasm) of Chlamydia trachomatis-infected epithelial cells. The overall intensity of the Raman spectrum is greatest in the inclusions and lowest in the cytoplasm in fixed cells. Difference spectra generated by normalizing to the intensity of the strong 1004 cm(-1) phenylalanine line show distinct differences among the three compartments. Most notably, the concentrations of adenine are greater in both the inclusions and the nucleus than in the cytoplasm, as seen by Raman microscopy. The source of the adenine was explored through a complementary approach, using two-photon microscopy imaging. Autofluorescence measurements of living, infected cells show that the adenine-containing molecules, NAD(P)H and FAD, are present mainly in the cytoplasm, suggesting that these molecules are not the source of the additional adenine signal in the nucleus and inclusions. Experiments of infected cells stained with a DNA-binding dye, Hoechst 33258, reveal that most of the DNA is present in the nucleus and the inclusions, suggesting that DNA/RNA is the main source of the additional Raman adenine signal in the nucleus and inclusions. Thus, Raman and two-photon microscopy are among the few non-invasive methods available to investigate cells infected with Chlamydia and, together, should also be useful for studying infection by other intracellular pathogens that survive within intracellular vacuoles.

  12. Autofluorescence imaging and magnification endoscopy

    Institute of Scientific and Technical Information of China (English)

    Monalisa Filip; Sevasti(t)a Iordache; Adrian S(a)ftoiu; Tudorel Ciurea

    2011-01-01

    It is well known that angiogenesis is critical in the transition from premalignant to malignant lesions.Consequently,early detection and diagnosis based on morphological changes to the microvessels are crucial.In the last few years,new imaging techniques which utilize the properties of light-tissue interaction have been developed to increase early diagnosis of gastrointestinal (GI)tract neoplasia.We analyzed several "red-flag" endoscopic techniques used to enhance visualization of the vascular pattern of preneoplastic and neoplastic lesions (e.g.trimodal imaging including autofluorescence imaging,magnifying endoscopy and narrow band imaging).These new endoscopic techniques provide better visualization of mucosal microsurface structure and microvascular architecture and may enhance the diagnosis and characterization of mucosal lesions in the GI tract.In the near future,it is expected that trimodal imaging endoscopy will be practiced as a standard endoscopy technique as it is quick,safe and accurate for making a precise diagnosis of gastrointestinal pathology,with an emphasis on the diagnosis of early GI tract cancers.Further large-scale randomized controlled trials comparing these modalities in different patient subpopulations are warranted before their endorsement in the routine practice of GI endoscopy.

  13. Optimization of the spectral design used to detect early carcinoma in the human tracheo-bronchial tree by autofluorescence imaging

    Science.gov (United States)

    Gabrecht, Tanja; Glanzmann, Thomas; Freitag, Lutz; Grosjean, Pierre; Weber, Bernd C.; Monnier, Philippe; van den Bergh, Hubert; Wagnieres, Georges A.

    2003-10-01

    The early detection and localization of bronchial cancer remains a challenging task. Autofluorescence bronchoscopy is emerging as a useful diagnostic tool with improved sensitivity and specificity. Evidence exists that the native fluorescence or autofluorescence of bronchial tissues changes when they turn dysplastic or to carcinoma in situ (CIS). Early lesions in the bronchi tend to show a decrease in autofluorescence in the green region of the spectrum when excited with violet light and a relative increase in the red region of the spectrum. Several endoscopic imaging devices relying on these optical properties of bronchial mucosa have been developed. An industrial endoscopic autofluorescence imaging system for the detection of early cancerous lesions in the bronchi has been developed in collaboration with the firm Richard Wolf Endoskope GmbH, Knittlingen (Germany; its performance has been evaluated in a previous clinical study. A second study, presented in this article, aims to optimize the spectral design of the device. Twenty-four lung cancer or high risk patients were enrolled in this study to assess the influence of additional backscattered red light on the tumor-to-healthy tissue contrast and to compare the effect of a narrow band violet excitation to a large band violet excitation. In our study we observed a three times higher contrast between cancer and healthy tissue, when backscattered red light was added to the violet excitation. The comparison between a narrow and a large band violet excitation indicated an increase of the tumor-to-healthy tissue contrast by the narrow band excitation.

  14. New design strategy for the two-photon active material based on push-pull substituted bisanthene molecule.

    Science.gov (United States)

    Chattopadhyaya, Mausumi; Alam, Md Mehboob; Chakrabarti, Swapan

    2011-03-31

    In the present work, we have critically examined the origin of strong two-photon transition probability of a donor-acceptor substituted bisanthene molecule that imitates a small piece of edge passivated (4, 4) graphene nanoribbon. In our calculations, we have considered -OMe, and -NH(2) as donors and -NO(2) as an acceptor. The one- and two-photon absorption parameters are evaluated using state-of-the-art linear and quadratic response theory, respectively, and all these calculations are carried out within the framework of time dependent density functional theory. To give a proper judgment on our findings, we have used the long-range corrected CAMB3LYP functional for all of the time dependent calculations. The present investigation reveals that the bisanthene molecule with three pairs of donor/acceptor moiety has a lower two-photon transition probability than that of a suitably designed bisanthene with only a single pair of donor/acceptor moiety. This in silico observation is consistent for all of the donor/acceptor moieties chosen in the present work. A comprehensive analysis at the two state model level of theory clearly offers us a verdict that by placing the donor/acceptor moiety in a suitable position of bisanthene, we can create a significant asymmetry in the electron density in the first excited state, which eventually leads to a significant difference in the ground and excited state dipole moment and is attributed to the higher two-photon transition probability of a particular bisanthene with a single pair of donor/acceptor moiety than bisanthene with three pairs of donor/acceptor.

  15. Clinical multiphoton tomography and clinical two-photon microendoscopy

    Science.gov (United States)

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

    2009-02-01

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

  16. Two-Photon Micromaser with Initial Atomic Coherence

    Institute of Scientific and Technical Information of China (English)

    SUN Wei-Hui; DU Si-De; CHEN Xiao-Shuang

    2005-01-01

    @@ We investigate the quantum dynamics ora two-photon micromaser pumped by atoms injected in the superpositionstate of the upper and intermediate levels. We simulate a master equation governing the system by the MonteCarlo wavefunction approach and analyse the steady-state behaviour as a function of the atomic transit time.The atomic coherence can effectively enhance the intensity and sub-Poissonian of the cavity field as comparedwith the atomic mixture. It is also discovered that the phase of the cavity field can be shifted by adjusting thedetuning between the atom and field. This result shows that it is possible to manipulate the phase of the cavityfield by detuning, due to atomic coherence.

  17. Whole brain imaging with Serial Two-Photon Tomography

    Directory of Open Access Journals (Sweden)

    Stephen P Amato

    2016-03-01

    Full Text Available Imaging entire mouse brains at submicron resolution has historically been a challenging undertaking and largely confined to the province of dedicated atlasing initiatives. The has limited systematic investigations into important areas of neuroscience, such as neural circuits, brain mapping and neurodegeneration. In this paper, we describe in detail Serial Two-Photon (STP tomography, a robust, reliable method for imaging entire brains with histological detail. We provide examples of how the basic methodology can be extended to other imaging modalities, such as optical coherence tomography, in order to provide unique contrast mechanisms. Furthermore we provide a survey of the research that STP tomography has enabled in the field of neuroscience, provide examples of how this technology enables quantitative whole brain studies, and discuss the current limitations of STP tomography-based approaches

  18. Two-photon assisted clock comparison to picosecond precision

    CERN Document Server

    Zhang, Shi-Wei; Yao, Yin-Ping; Wan, Ren-Gang; Zhang, Tong-Yi

    2015-01-01

    We have experimentally demonstrated a clock comparison scheme utilizing time-correlated photon pairs generated from the spontaneous parametric down conversion process of a laser pumped beta-barium borate crystal. The coincidence of two-photon events are analyzed by the cross correlation of the two time stamp sequences. Combining the coarse and fine part of the time differences at different resolutions, a 64 ps precision for clock synchronization has been realized. We also investigate the effects of hardware devices used in the system on the precision of clock comparison. The results indicate that the detector's time jitter and the background noise will degrade the system performance. With this method, comparison and synchronization of two remote clocks could be implemented with a precision at the level of a few tens of picoseconds.

  19. Measurement of bottom quark production in two photon collisions

    CERN Document Server

    Saremi, Sepehr

    2001-01-01

    The cross section for bottom quark production in two-photon collisions, sigma( e+e- → e+e- bb¯X), is measured for the first time. The measurement is performed with the L3 detector at the Large Electron Positron (LEP) collider at the European Center for Nuclear and Particle Physics (CERN). The data corresponds to 410 pb-1 taken at center-of-mass energies from 189 GeV to 202 GeV. Hadrons containing a bottom quark are identified by detecting electrons or muons from their semi-leptonic decays. The measured cross section is in excess of the Next to Leading Order QCD prediction by a factor of three.

  20. Two-photon transition form factor of c ¯ quarkonia

    Science.gov (United States)

    Chen, Jing; Ding, Minghui; Chang, Lei; Liu, Yu-xin

    2017-01-01

    The two-photon transition of c ¯c quarkonia are studied within a covariant approach based on the consistent truncation scheme of the quantum chromodynamics Dyson-Schwinger equation for the quark propagator and the Bethe-Salpeter equation for the mesons. We find the decay widths of ηc→γ γ and χc 0 ,2→γ γ in good agreement with experimental data. The obtained transition form factor of ηc→γ γ* for a wide range of spacelike photon-momentum-transfer squared is also in agreement with the experimental findings of the BABAR experiment. As a by-product, the decay widths of ηb,χb 0 ,2→γ γ and the transition form factor of ηb,χc 0 ,b 0→γ γ* are predicted, which await experimental testing.

  1. Human skin auto-fluorescence decay as a function of irradiance and skin type

    Science.gov (United States)

    Debreczeny, Martin P.; Bates, Rebecca; Fitch, Rick M.; Galen, Karen P.; Ge, Jiajia; Dorshow, Richard B.

    2011-03-01

    The aim of this work was to establish measurement conditions under which endogenous skin fluorescence ("auto-fluorescence") is relatively invariant, so that changes in exogenous agents can be accurately determined. Fluorescence emission was measured on the volar forearm of 36 subjects, chosen to be equally representative of all 6 Fitzpatrick skin types. All subjects were exposed to approximately 40 minutes of optical excitation at 450 and 500 nm with 4 irradiances between 0.3 and 9 mW/cm2. Both non-optically-induced (e.g. tissue settling and fluctuation) and optically-induced variations were observed in the measured fluorescence and mechanisms explaining these effects are proposed. The optically-induced auto-fluorescence decay was independent of skin type when excited at 450 nm, but significantly dependent on skin type when excited at 500 nm. Further, the extent of decay over time was linearly related to irradiance at 500 nm, but at 450 nm was non-linear, with the extent of decay rolling off between 2 and 9 mW/cm2. In order to maintain the auto-fluorescence signal within 95% of its original value over a 30 minute period, the excitation at 450 nm would need to be limited to 1.5 mW/cm2, while excitation at 500 nm should be limited to 5 mW/cm2.

  2. Enhanced two photon fluorescence microfluidic sensor based on dual cladding photonic-crystal fiber

    Science.gov (United States)

    Amitonova, Lyubov; Fedotov, Ilya; Fedotov, Andrey; Zheltikov, Aleksei

    2012-11-01

    The architecture of photonic-crystal fibers (PCFs) suggests a variety of strategies for optical sensing. A combination of TPA approaches with capabilities of fiber-optic probes offers numerous advantages, suggesting a convenient format for beam delivery, facilitating manipulation of excitation radiation, and allowing this excitation to be applied locally and selectively. In this work, we show that a PCF with a special design can realize different protocols of optical sensing, simultaneously serving, whenever necessary, for the collection and on-line monitoring of liquid-phase samples. Specially designed PCF is shown to substantially increase the guided-wave luminescent response from molecules excited through two-photon absorption (TPA) by femtosecond near-infrared laser pulses. Biophotonic implications of this waveguide TPL-response enhancement include fiber-format solutions for online monitoring of drug delivery and drug activation, interrogation of neural activity, biosensing, endoscopy, and locally controlled singlet oxygen generation in photodynamic therapy. This work was supported by the Russian Foundation for Basic Research, project 11-04-12185-ofi-m.

  3. Broadband Two-Photon Absorption Characteristics of Highly Photostable Fluorenyl-Dicyanoethylenylated [60]Fullerene Dyads

    Directory of Open Access Journals (Sweden)

    Seaho Jeon

    2016-05-01

    Full Text Available We synthesized four C60-(light-harvesting antenna dyads C60 (>CPAF-Cn (n = 4, 9, 12, or 18 1-Cn for the investigation of their broadband nonlinear absorption effect. Since we have previously demonstrated their high function as two-photon absorption (2PA materials at 1000 nm, a different 2PA wavelength of 780 nm was applied in the study. The combined data taken at two different wavelength ranges substantiated the broadband characteristics of 1-Cn. We proposed that the observed broadband absorptions may be attributed by a partial π-conjugation between the C60 > cage and CPAF-Cn moieties, via endinitrile tautomeric resonance, giving a resonance state with enhanced molecular conjugation. This transient state could increase its 2PA and excited-state absorption at 800 nm. In addition, a trend of concentration-dependent 2PA cross-section (σ2 and excited-state absorption magnitude was detected showing a higher σ value at a lower concentration that was correlated to increasing molecular separation with less aggregation for dyads C60(>CPAF-C18 and C60(>CPAF-C9, as better 2PA and excited-state absorbers.

  4. Singlet oxygen induced advanced glycation end-product photobleaching of in vivo human fingertip autofluorescence

    Science.gov (United States)

    Deng, Bin; Simental, Anabel; Lutz, Patrick; Shaheen, George; Chaiken, Joseph

    2012-01-01

    Nonenzymatic glycation and oxidation of ubiquitous proteins in vivo leads to irreversible formation of advanced glycation end products (AGEs). Due to their relatively long half life and low clearance rate AGEs tend to accumulate within static tissues and the circulatory system. Spectra obtained using 830 nm near-infrared (NIR) excitation suggest that the so-called "autofluorescence" from all tissues has a finite number of sources but the fact that senior and diabetic subjects produce more than other members of the general population suggests that a significant portion of the total autofluorescence from all sources originates from AGEs. Using pentosidine generated in a reaction mixture as described by Monnier as representative, an in vitro study unveiled very similar fluorescence and photobleaching pattern as observed for autofluorescence in vivo. A series of oxygen, air and argon purging experiments on the pentosidine-generating reaction mixture suggests that pentosidine is a singlet oxygen sensitizer and secondary reactions between the pentosidine itself and/or other fluorophores and the photosensitized singlet oxygen explain the observed photobleaching. Ab initio Gaussian calculations on pentosidine reveal the existence of low-lying triplet excited states required for the sensitization of ground state oxygen. A commercially available product known as singlet oxygen sensor green (SOSG) that specifically serves as a singlet oxygen detection reagent confirms the generation of singlet oxygen from NIR irradiated pentosidine trimixture. This study provides one definite chemical mechanism for understanding in vivo human skin autofluorescence and photobleaching.

  5. Autofluorescence-free in vivo multicolor imaging using upconversion fluoride nanocrystals.

    Science.gov (United States)

    Tian, Zhen; Chen, Guanying; Li, Xiang; Liang, Huijuan; Li, Yuanshi; Zhang, Zhiguo; Tian, Ye

    2010-07-01

    Non-invasive fluorescence imaging is an important technique in biology. However, detection of traditional biomarker emissions is accompanied by a high background signal. In this study we examined whether upconversion sodium yttrium fluoride (NaYF(4)) nanocrystals were suitable for autofluorescence-free multicolor fluorescence imaging in a living animal. Tissue autofluorescence was induced with a 405 nm light source, then rats were subjected to injection of fluorescein isothiocyanate (FITC), cadmium selenide/zinc sulfide (CdSe/ZnS) quantum dots (QDs), or NaYF(4):ytterbium/thulium (Yb(3+)/Tm(3+)), NaYF(4):Yb(3+)/holmium (Ho(3+)), and NaYF(4):Yb(3+)/Ho(3+)/cerium (Ce(3+)) nanocrystals. Imaging with NaYF(4) nanocrystals (974 nm laser) completely removed the high tissue autofluorescence, in marked contrast to imaging with FITC and QDs (405 nm light). Optical imaging experiments demonstrated that multiple biological targets and organs could be imaged at the same time using multicolor NaYF(4) upconversion nanocrystals under a single excitation wavelength (974 nm). These data demonstrated the proof-of-principle that autofluorescence-free multicolor imaging using near-infrared to visible upconversion of NaYF(4) nanocrystals excited by laser can be performed in a living animal.

  6. Theoretical analysis on two-photon absorption spectroscopy in a confined four-level atomic system

    Institute of Scientific and Technical Information of China (English)

    Yuanyuan Li; Jintao Bai; Li Li; Yanpeng Zhang; Xun Hou

    2009-01-01

    We investigate theoretically two-photon absorption spectroscopy modified by a control field in a confined Y-type four-level system. Dicke-narrowing effect occurs both in two-photon absorption lines and the dips of transparency against two-photon absorption due to enhanced contribution of slow atoms. We also find that the suppression and the enhancement of two-photon absorption can be modified by changing the strength of the control field and the detuning of three laser fields. This control of two-photon absorption may have some applications in information processing and optical devices.

  7. Two-dimensional imaging of molecular hydrogen in H2-air diffusion flames using two-photon laser-induced fluorescence

    Science.gov (United States)

    Lempert, W.; Kumar, V.; Glesk, I.; Miles, R.; Diskin, G.

    1991-01-01

    The use of a tunable ArF laser at 193.26 nm to record simultaneous single-laser-shot, planar images of molecular hydrogen and hot oxygen in a turbulent H2-air diffusion flame. Excitation spectra of fuel and oxidant-rich flame zones confirm a partial overlap of the two-photon H2 and single-photon O2 Schumann-Runge absorption bands. UV Rayleigh scattering images of flame structure and estimated detection limits for the H2 two-photon imaging are also presented.

  8. Acetylene bridged porphyrin-monophthalocyaninato ytterbium(III) hybrids with strong two-photon absorption and high singlet oxygen quantum yield.

    Science.gov (United States)

    Ke, Hanzhong; Li, Wenbin; Zhang, Tao; Zhu, Xunjin; Tam, Hoi-Lam; Hou, Anxin; Kwong, Daniel W J; Wong, Wai-Kwok

    2012-04-21

    Several acetylene bridged porphyrin-monophthalocyaninato ytterbium(III) hybrids, PZn-PcYb, PH(2)-PcYb and PPd-PcYb, have been prepared and characterized by (1)H and (31)P NMR, mass spectrometry, and UV-vis spectroscopy. Their photophysical and photochemical properties, especially the relative singlet oxygen ((1)O(2)) quantum yields and the two-photon absorption cross-section (σ(2)), were investigated. These three newly synthesized compounds exhibited very large σ(2) values and substantial (1)O(2) quantum yields upon photo-excitation, making them potential candidates as one- and two-photon photodynamic therapeutic agents.

  9. In vivo monitoring of intracellular chloroplast movements in intact leaves of C4 plants using two-photon microscopy.

    Science.gov (United States)

    Ryu, Jeongeun; Nam, Hyoseok; Kim, Hae Koo; Joo, Yongjoon; Lee, Sang Joon; Kim, Ki Hean

    2014-10-01

    Dynamic changes in the spatial distribution of chloroplasts are essential for optimizing photosynthetic capacity under changing light conditions. Light-induced movement of chloroplasts has been widely investigated, but most studies were conducted on isolated tissues or protoplasts. In this study, a two-photon microscopy (TPM) system was adapted to monitor the intracellular 3-dimensional (3D) movements of chloroplasts in intact leaves of plants during dark to light transitions. The TPM imaging was based on autofluorescence of chlorophyll generated by a femto-second Ti:Sapphire laser. All chloroplasts did not exhibit the same motion in response to irradiation variation. In the sub-epidermal mesophyll cells, chloroplasts generally moved away from the surface following blue light treatment, however many chloroplasts did not show any movement. Such spatial heterogeneity in chloroplast motility underlines the importance of monitoring intracellular orientation and movement of individual chloroplasts across intact leaves. Our investigation shows that the 3D imaging of chloroplasts using TPM can help to understand the changes in local photosynthetic capacity in intact leaves under changing environmental conditions.

  10. Comparison of reflectance confocal microscopy and two-photon second harmonic generation microscopy in fungal keratitis rabbit model ex vivo

    Science.gov (United States)

    Lee, Jun Ho; Lee, Seunghun; Yoon, Calvin J.; Park, Jin Hyoung; Tchah, Hungwon; Kim, Myoung Joon; Kim, Ki Hean

    2016-01-01

    Fungal keratitis is an infection of the cornea by fungal pathogens. Diagnosis methods based on optical microscopy could be beneficial over the conventional microbiology method by allowing rapid and non-invasive examination. Reflectance confocal microscopy (RCM) and two-photon second harmonic generation microscopy (TPSHGM) have been applied to pre-clinical or clinical studies of fungal keratitis. In this report, RCM and TPSHGM were characterized and compared in the imaging of a fungal keratitis rabbit model ex vivo. Fungal infection was induced by using two strains of fungi: aspergillus fumigatus and candida albicans. The infected corneas were imaged in fresh condition by both modalities sequentially and their images were analyzed. Both RCM and TPSHGM could detect both fungal strains within the cornea based on morphology: aspergillus fumigatus had distinctive filamentous structures, and candida albicans had round structures superficially and elongated structures in the corneal stroma. These imaging results were confirmed by histology. Comparison between RCM and TPSHGM showed several characteristics. Although RCM and TPSHGM images had good correlation each other, their images were slightly different due to difference in contrast mechanism. RCM had relatively low image contrast with the infected turbid corneas due to high background signal. TPSHGM visualized cells and collagen in the cornea clearly compared to RCM, but used higher laser power to compensate low autofluorescence. Since these two modalities provide complementary information, combination of RCM and TPSHGM would be useful for fungal keratitis detection by compensating their weaknesses each other. PMID:26977371

  11. Two-Photon Ghost Image and Interference-Diffraction

    Science.gov (United States)

    Shih, Y. H.; Sergienko, A. V.; Pittman, T. B.; Strekalov, D. V.; Klyshko, D. N.

    1996-01-01

    One of the most surprising consequences of quantum mechanics is entanglement of two or more distance particles. The two-particle entangled state was mathematically formulated by Schrodinger. Based on this unusual quantum behavior, EPR defined their 'physical reality' and then asked the question: 'Can Quantum-Mechanical Description of Physical Reality Be Considered Complete?' One may not appreciate EPR's criterion of physical reality and insist that 'no elementary quantum phenomenon is a phenomenon until it is a recorded phenomenon'. Optical spontaneous parametric down conversion (SPDC) is the most effective mechanism to generate an EPR type entangled two-photon state. In SPDC, an optical beam, called the pump, is incident on a birefringent crystal. The pump is intense enough so that nonlinear effects lead to the conversion of pump photons into pairs of photons, historically called signal and idler. Technically, the SPDC is said to be type-1 or type-2, depending on whether the signal and idler beams have parallel or orthogonal polarization. The SPDC conversion efficiency is typically on the order of 10(exp -9) to 10(exp -11), depending on the SPDC nonlinear material. The signal and idler intensities are extremely low, only single photon detection devices can register them. The quantum entanglement nature of SPDC has been demonstrated in EPR-Bohm experiments and Bell's inequality measurements. The following two experiments were recently performed in our laboratory, which are more closely related to the original 1935 EPR gedankenezperiment. The first experiment is a two-photon optical imaging type experiment, which has been named 'ghost image' by the physics community. The signal and idler beams of SPDC are sent in different directions, so that the detection of the signal and idler photons can be performed by two distant photon counting detectors. An aperture object (mask) is placed in front of the signal photon detector and illuminated by the signal beam through a

  12. Hyperspectral laser-induced autofluorescence imaging of dental caries

    Science.gov (United States)

    Bürmen, Miran; Fidler, Aleš; Pernuš, Franjo; Likar, Boštjan

    2012-01-01

    Dental caries is a disease characterized by demineralization of enamel crystals leading to the penetration of bacteria into the dentine and pulp. Early detection of enamel demineralization resulting in increased enamel porosity, commonly known as white spots, is a difficult diagnostic task. Laser induced autofluorescence was shown to be a useful method for early detection of demineralization. The existing studies involved either a single point spectroscopic measurements or imaging at a single spectral band. In the case of spectroscopic measurements, very little or no spatial information is acquired and the measured autofluorescence signal strongly depends on the position and orientation of the probe. On the other hand, single-band spectral imaging can be substantially affected by local spectral artefacts. Such effects can significantly interfere with automated methods for detection of early caries lesions. In contrast, hyperspectral imaging effectively combines the spatial information of imaging methods with the spectral information of spectroscopic methods providing excellent basis for development of robust and reliable algorithms for automated classification and analysis of hard dental tissues. In this paper, we employ 405 nm laser excitation of natural caries lesions. The fluorescence signal is acquired by a state-of-the-art hyperspectral imaging system consisting of a high-resolution acousto-optic tunable filter (AOTF) and a highly sensitive Scientific CMOS camera in the spectral range from 550 nm to 800 nm. The results are compared to the contrast obtained by near-infrared hyperspectral imaging technique employed in the existing studies on early detection of dental caries.

  13. Evidence for strong electron correlations in graphene molecular fragments: Theory and experiments on two-photon absorptions

    Science.gov (United States)

    Aryanpour, Karan; Roberts, Adam; Sandhu, Arvinder; Shukla, Alok; Mazumdar, Sumit

    2013-03-01

    Historically, the occurrence of the lowest two-photon state below the optical one-photon state in linear polyenes, polyacetylenes and polydiacetylenes provided the strongest evidence for strong electron correlations in these linear π-conjugated systems. We demonstrate similar behavior in several molecular fragments of graphene with D6 h symmetry, theoretically and experimentally. Theoretically, we have calculated one versus two-photon absorptions in coronene, two different hexabenzocoronenes and circumcoronene, within the Pariser-Parr-Pople π-electron Hamiltonian using high order configuration interaction. Experimentally, we have performed z-scan measurements using a white light super-continuum source on coronene and hexa-peri-hexabenzocoronene to determine frequency-dependent two-photon absorption coefficients, for comparison to the ground state absorptions. Excellent agreement between experiment and theory in our work gives strong evidence for significant electron correlations between the π-electrons in the graphene molecular fragments. We particularly benchmark high order electron-hole excitations in graphene fragments as a key element behind the agreement between theory and experiment in this work. We acknowledge NSF-CHE-1151475 grant as our funding source.

  14. In Vivo Non Linear Optical (NLO) Imaging in Live Rabbit Eyes Using the Heidelberg Two-Photon Laser Ophthalmoscope

    Science.gov (United States)

    Hao, Ming; Flynn, Kevin; Nien-Shy, Chyong; Jester, Bryan E.; Winkler, Moritz; Brown, Donald J.; La Schiazza, Olivier; Bille, Josef; Jester, James V.

    2010-01-01

    Imaging of non-linear optical (NLO) signals generated from the eye using ultrafast pulsed lasers has been limited to the study of ex vivo tissues because of the use of conventional microscopes with slow scan speeds. The purpose of this study was to evaluate the ability of a novel, high scan rate ophthalmoscope to generate NLO signals using an attached femtosecond laser. NLO signals were generated and imaged in live, anesthetized albino rabbits using a newly designed Heidelberg Two-Photon Laser Ophthalmoscope with attached 25 mW femtosecond laser having a central wavelength of 780 nm, pulsewidth of 75 fs, and a repetition rate of 50 MHz. To assess two-photon excited fluorescent (TPEF) signal generation, cultured rabbit corneal fibroblasts (RCF) were first labeled by Blue-green fluorescent FluoSpheres (1 μm diameter) and then cells were micro-injected into the central cornea. Clumps of RCF cells could be detected by both reflectance and TPEF imaging at 6 hours after injection. By 6 days, RCF containing fluorescent microspheres confirmed by TPEF showed a more spread morphology and had migrated from the original injection site. Overall, this study demonstrates the potential of using NLO microscopy to sequentially detect TPEF signals from live, intact corneas. We conclude that further refinement of the Two-photon laser Ophthalmoscope should lead to the development of an important, new clinical instrument capable of detecting NLO signals from patient corneas. PMID:20558159

  15. Theory of deep ultraviolet generation at maximum coherence assisted by Stark-chirped two-photon resonance

    CERN Document Server

    Myslivets, S A; Kimberg, V V; George, T F; George, Thomas F.

    2003-01-01

    A scheme is analyzed for effcient generation of vacuum ultraviolet radiation through four-wave mixing processes assisted by the technique of Stark-chirped rapid adiabatic passage. These opportunities are associated with pulse excitation of laddertype short-wavelength two-photon atomic or molecular transitions so that relaxation processes can be neglected. In this three-laser technique, a delayed-pulse of strong oR-resonant infrared radiation sweeps the laser-induced Stark-shift of a two-photon transition in a such way that facilitates robust maximum two-photon coherence induced by the first ultraviolet laser. A judiciously delayed third pulse scatters at this coherence and generates short-wavelength radiation. A theoretical analysis of these problems based on the density matrix is performed. A numerical model is developed to carry out simulations of a typical experiment. The results illustrate a behavior of populations, coherence and generated radiation along the medium as well as opportunities of effcient ge...

  16. Autocorrelation measurement of femtosecond laser pulses based on two-photon absorption in GaP photodiode

    Energy Technology Data Exchange (ETDEWEB)

    Chong, E. Z.; Watson, T. F.; Festy, F., E-mail: frederic.festy@kcl.ac.uk [Biomaterials, Biomimetics and Biophotonics Division, King' s College London—Dental Institute, SE1 9RT London (United Kingdom)

    2014-08-11

    Semiconductor materials which exhibit two-photon absorption characteristic within a spectral region of interest can be useful in building an ultra-compact interferometric autocorrelator. In this paper, we report on the evidence of a nonlinear absorption process in GaP photodiodes which was exploited to measure the temporal profile of femtosecond Ti:sapphire laser pulses with a tunable peak wavelength above 680 nm. The two-photon mediated conductivity measurements were performed at an average laser power of less than a few tenths of milliwatts. Its suitability as a single detector in a broadband autocorrelator setup was assessed by investigating the nonlinear spectral sensitivity bandwidth of a GaP photodiode. The highly favourable nonlinear response was found to cover the entire tuning range of our Ti:sapphire laser and can potentially be extended to wavelengths below 680 nm. We also demonstrated the flexibility of GaP in determining the optimum compensation value of the group delay dispersion required to restore the positively chirped pulses inherent in our experimental optical system to the shortest pulse width possible. With the rise in the popularity of nonlinear microscopy, the broad two-photon response of GaP and the simplicity of this technique can provide an alternative way of measuring the excitation laser pulse duration at the focal point of any microscopy systems.

  17. Pressure broadening of atomic oxygen two-photon absorption laser induced fluorescence

    Science.gov (United States)

    Marinov, Daniil; Drag, Cyril; Blondel, Christophe; Guaitella, Olivier; Golda, Judith; Klarenaar, Bart; Engeln, Richard; Schulz-von der Gathen, Volker; Booth, Jean-Paul

    2016-12-01

    Atomic oxygen, considered to be a determining reactant in plasma applications at ambient pressure, is routinely detected by two-photon absorption laser induced fluorescence (TALIF). Here, pressure broadening of the (2p 4 3 P 2  →  3p 3 P J=0,1,2) two-photon transition in oxygen atoms was investigated using a high-resolution TALIF technique in normal and Doppler-free configurations. The pressure broadening coefficients determined were {γ{{\\text{O}2}}}   =  0.40  ±  0.08  cm-1/bar for oxygen molecules and {γ\\text{He}}   =  0.46  ±  0.03 cm-1/bar for helium atoms. These correspond to pressure broadening rate constants k\\text{PB}{{\\text{O}2}}   =  9 · 10-9 cm3 s-1 and k\\text{PB}\\text{He}   =  4 · 10-9 cm3 s-1, respectively. The well-known quenching rate constants of O(3p 3 P J ) by O2 and He are at least one order of magnitude smaller, which signifies that non-quenching collisions constitute the main line-broadening mechanism. In addition to providing new insights into collisional processes of oxygen atoms in electronically excited 3p 3 P J state, reported pressure broadening parameters are important for quantification of oxygen TALIF line profiles when both collisional and Doppler broadening mechanisms are important. Thus, the Doppler component (and hence the temperature of oxygen atoms) can be accurately determined from high resolution TALIF measurements in a broad range of conditions.

  18. Two-photon photoemission study of the coverage-dependent electronic structure of chemisorbed alkali atoms on a Ag(111) surface.

    Science.gov (United States)

    Wang, Lei-Ming; Sametoglu, Vahit; Winkelmann, Aimo; Zhao, Jin; Petek, Hrvoje

    2011-09-01

    We report a systematic investigation of the electronic structure of chemisorbed alkali atoms (Li-Cs) on a Ag(111) surface by two-photon photoemission spectroscopy. Angle-resolved two-photon photoemission spectra are obtained for 0-0.1 monolayer coverage of alkali atoms. The interfacial electronic structure as a function of periodic properties and the coverage of alkali atoms is observed and interpreted assuming ionic adsorbate/substrate interaction. The energy of the alkali atom σ-resonance at the limit of zero coverage is primarily determined by the image charge interaction, whereas at finite alkali atom coverages, it follows the formation of a dipolar surface field. The coverage- and angle-dependent two-photon photoemission spectra provide information on the photoinduced charge-transfer excitation of adsorbates on metal surfaces. This work complements the previous work on alkali/Cu(111) chemisorption [Phys. Rev. B 2008, 78, 085419].

  19. Two-photon polymerization for fabrication of biomedical devices

    Science.gov (United States)

    Ovsianikov, Aleksandr; Doraiswamy, Anand; Narayan, R.; Chichkov, B. N.

    2007-01-01

    Two-photon polymerization (2PP) is a novel technology which allows the fabrication of complex three-dimensional (3D) microstructures and nanostructures. The number of applications of this technology is rapidly increasing; it includes the fabrication of 3D photonic crystals [1-4], medical devices, and tissue scaffolds [5-6]. In this contribution, we discuss current applications of 2PP for microstructuring of biomedical devices used in drug delivery. While in general this sector is still dominated by oral administration of drugs, precise dosing, safety, and convenience are being addressed by transdermal drug delivery systems. Currently, main limitations arise from low permeability of the skin. As a result, only few types of pharmacological substances can be delivered in this manner [7]. Application of microneedle arrays, whose function is to help overcome the barrier presented by the epidermis layer of the skin, provides a very promising solution. Using 2PP we have fabricated arrays of hollow microneedles with different geometries. The effect of microneedle geometry on skin penetration is examined. Our results indicate that microneedles created using 2PP technique are suitable for in vivo use, and for integration with the next generation of MEMS- and NEMS-based drug delivery devices.

  20. Review of two-photon exchange in electron scattering

    Energy Technology Data Exchange (ETDEWEB)

    J. Arrington, P. G. Blunden, W. Melnitchouk

    2011-10-01

    We review the role of two-photon exchange (TPE) in electron-hadron scattering, focusing in particular on hadronic frameworks suitable for describing the low and moderate Q^2 region relevant to most experimental studies. We discuss the effects of TPE on the extraction of nucleon form factors and their role in the resolution of the proton electric to magnetic form factor ratio puzzle. The implications of TPE on various other observables, including neutron form factors, electroproduction of resonances and pions, and nuclear form factors, are summarized. Measurements seeking to directly identify TPE effects, such as through the angular dependence of polarization measurements, nonlinear epsilon contributions to the cross sections, and via e+p to e-p cross section ratios, are also outlined. In the weak sector, we describe the role of TPE and gamma-Z interference in parity-violating electron scattering, and assess their impact on the extraction of the strange form factors of the nucleon and the weak charge of the proton.

  1. Higgs decay into two photons in a warped extra dimension

    Energy Technology Data Exchange (ETDEWEB)

    Hahn, Juliane; Hoerner, Clara; Malm, Raoul; Novotny, Kristiane; Schmell, Christoph [Johannes Gutenberg University, PRISMA Cluster of Excellence and Mainz Institute for Theoretical Physics, Mainz (Germany); Neubert, Matthias [Johannes Gutenberg University, PRISMA Cluster of Excellence and Mainz Institute for Theoretical Physics, Mainz (Germany); Cornell University, Department of Physics, LEPP, Ithaca, NY (United States)

    2014-05-15

    A detailed five-dimensional calculation of the Higgs-boson decay into two photons is performed in both the minimal and the custodially protected Randall-Sundrum (RS) model, where the Standard Model (SM) fields propagate in the bulk and the scalar sector lives on or near the IR brane. It is explicitly shown that the R{sub ξ} gauge invariance of the sum of diagrams involving bosonic fields in the SM also applies to the case of these RS scenarios. An exact expression for the h → γγ amplitude in terms of the five-dimensional (5D) gauge-boson and fermion propagators is presented, which includes the full dependence on the Higgs-boson mass. Closed expressions for the 5D W-boson propagators in theminimal and the custodial RS model are derived, which are valid to all orders in v{sup 2}/M{sup 2}{sub KK}. In contrast to the fermion case, the result for the bosonic contributions to the h → γγ amplitude is insensitive to the details of the localization of the Higgs profile on or near the IR brane. The various RS predictions for the rate of the pp → h → γγ process are compared with the latest LHC data, and exclusion regions for the RS model parameters are derived. (orig.)

  2. Two-Photon-Absorption Scheme for Optical Beam Tracking

    Science.gov (United States)

    Ortiz, Gerardo G.; Farr, William H.

    2011-01-01

    A new optical beam tracking approach for free-space optical communication links using two-photon absorption (TPA) in a high-bandgap detector material was demonstrated. This tracking scheme is part of the canonical architecture described in the preceding article. TPA is used to track a long-wavelength transmit laser while direct absorption on the same sensor simultaneously tracks a shorter-wavelength beacon. The TPA responsivity was measured for silicon using a PIN photodiode at a laser beacon wavelength of 1,550 nm. As expected, the responsivity shows a linear dependence with incident power level. The responsivity slope is 4.5 x 10(exp -7) A/W2. Also, optical beam spots from the 1,550-nm laser beacon were characterized on commercial charge coupled device (CCD) and complementary metal-oxide semiconductor (CMOS) imagers with as little as 13.7 microWatts of optical power (see figure). This new tracker technology offers an innovative solution to reduce system complexity, improve transmit/receive isolation, improve optical efficiency, improve signal-to-noise ratio (SNR), and reduce cost for free-space optical communications transceivers.

  3. Two-Photon-Exchange Effects and $\\Delta(1232)$ Deformation

    CERN Document Server

    Zhou, Hai-Qing

    2016-01-01

    The two-photon-exchange (TPE) contribution in $ep\\rightarrow ep\\pi ^0$ with $W=M_{\\Delta}$ and small $Q^2$ is calculated and its corrections to the ratios of electromagnetic transition form factors $R_{EM} = E_{1+}^{(3/2)}/M_{1+}^{(3/2)} $ and $R_{SM} = S_{1+}^{(3/2)}/M_{1+}^{(3/2)}$, are analysed. A simple hadronic model is used to estimate the TPE amplitude. Two phenomenological models, MAID2007 and SAID, are used to approximate the full $ep\\rightarrow ep\\pi ^0$ cross sections which contain both the TPE and the one-photon-exchange (OPE) contributions. The genuine the OPE amplitude is then extracted from an integral equation by iteration. We find that the TPE contribution is not sensitive to whether MAID or SAID is used as input in the region with $Q^2<2$ GeV$^2$. It gives small correction to $R_{EM}$ while for $R_{SM}$, the correction is about -10\\% at small $\\epsilon$ and about $1\\%$ at large $\\epsilon$ for $Q^2\\approx2.5$ GeV$^2$. The large correction from TPE at small $\\epsilon$ must be included in th...

  4. Time-resolved two-photon photoemission from metal surfaces

    CERN Document Server

    Weinelt, M

    2002-01-01

    The Rydberg-like series of image-potential states is a prototype system for loosely bound electrons at a metal surface. The electronic structure and the femtosecond dynamics of these states is studied by high-resolution energy-and time-resolved two-photon photoemission spectroscopy. The electron trapped in the image potential moves virtually freely laterally to the surface where it is subject to inelastic and quasielastic scattering processes which cause decay of population and phase relaxation. The influence of surface corrugation on these processes has been investigated for adsorbates on Cu(001) and stepped Cu(117) and Cu(119) surfaces which are vicinal to Cu(001). The dynamics depend on both the distance of the electron in front of the surface and the parallel momentum. For CO molecules on Cu(001) inelastic scattering into bulk states and adsorbate-induced resonances determine the decay rate. For small numbers of Cu adatoms on Cu(001) and the vicinal surfaces the decay rate of image-potential states is sig...

  5. Simultaneous control of emission localization and two-photon absorption efficiency in dissymmetrical chromophores

    Energy Technology Data Exchange (ETDEWEB)

    Tretiak, Sergei [Los Alamos National Laboratory

    2009-01-01

    The aim of the present work is to demonstrate that combined spectral tuning of fluorescence and two-photon absorption (TPA) properties of multipolar chromophores can be achieved by introduction of slight electronic chemical dissymmetry. In that perspective, two novel series of structurally related chromophores have been designed and studied: a first series based on rod-like quadrupolar chromophores bearing different electron-donating (D) end groups and a second series based on three-branched octupolar chromophores built from a trigonal donating moiety and bearing various acceptor (A) peripheral groups. The influence of the electronic dissymmetry is investigated by combined experimental and theoretical studies of the linear and nonlinear optical properties of dissymmetric chromophores compared to their symmetrical counterparts. In both types of systems (i.e. quadrupoles and octupoles) experiments and theory reveal that excitation is essentially delocalized and that excitation involves synchronized charge redistribution between the different D and A moieties within the multipolar structure (i.e. concerted intramolecular charge transfer). In contrast, the emission stems only from a particular dipolar subunit bearing the strongest D or A moieties due to fast excitation localization after excitation prior to emission. Hence control of emission characteristics (polarization and emission spectrum) in addition to localization can be achieved by controlled introduction of electronic dissymmetry (i.e. replacement of one of the D or A end-groups by a slightly stronger D{prime} or A{prime} units). Interestingly dissymmetrical functionalization of both quadrupolar and octupolar compounds does not lead to significant loss in TPA responses and can even be beneficial due to the spectral broadening and peak position tuning that it allows. This study thus reveals an original molecular engineering route strategy allowing major TPA enhancement in multipolar structures due to concerted

  6. Probing Electron-Phonon Interaction through Two-Photon Interference in Resonantly Driven Semiconductor Quantum Dots

    Science.gov (United States)

    Reigue, Antoine; Iles-Smith, Jake; Lux, Fabian; Monniello, Léonard; Bernard, Mathieu; Margaillan, Florent; Lemaitre, Aristide; Martinez, Anthony; McCutcheon, Dara P. S.; Mørk, Jesper; Hostein, Richard; Voliotis, Valia

    2017-06-01

    We investigate the temperature dependence of photon coherence properties through two-photon interference (TPI) measurements from a single quantum dot (QD) under resonant excitation. We show that the loss of indistinguishability is related only to the electron-phonon coupling and is not affected by spectral diffusion. Through these measurements and a complementary microscopic theory, we identify two independent separate decoherence processes, both of which are associated with phonons. Below 10 K, we find that the relaxation of the vibrational lattice is the dominant contribution to the loss of TPI visibility. This process is non-Markovian in nature and corresponds to real phonon transitions resulting in a broad phonon sideband in the QD emission spectra. Above 10 K, virtual phonon transitions to higher lying excited states in the QD become the dominant dephasing mechanism, this leads to a broadening of the zero phonon line, and a corresponding rapid decay in the visibility. The microscopic theory we develop provides analytic expressions for the dephasing rates for both virtual phonon scattering and non-Markovian lattice relaxation.

  7. In situ imaging of the mouse cochlea using two-photon microscopy

    Science.gov (United States)

    Yang, Xin; Pu, Ye; Psaltis, Demetri; Stankovic, Konstantina M.

    2013-04-01

    Intracochlear imaging is of great interest clinically because cochlea is the central organ of hearing. However, intracochlear imaging is technologically challenging due to the cochlea's small size and encasement in bone. The state-of- the-art imaging techniques are not adequate for high resolution cellular imaging to establish diagnosis without destroying the cochlea. We report in situ imaging of intact mouse cochlea using endogenous two-photon excitation fluorescence (TPEF) as the contrast mechanism. TPEF eliminates the need for exogenous labeling and eradicating the staining-induced artifacts. We used a natural, membranous opening into the cochlea, the round window, as the optical access to reach the organ of Corti, requiring no additional slicing or opening. Our approach provides the maximum non-invasiveness in the imaging process. TPEF exhibits strong contrast allowing deep imaging of mouse cochlea with cellular and even subcellular resolution. Inner hair cell, outer hair cell and supporting cell are clearly identifiable in TPEF images. Distinct morphological differences are observed between healthy and noise-exposed cochleae, allowing detection of specific, noise-induced pathologic changes. The TPEF images taken through the round window are correlated with the whole mount sections, verifying their reliability. Compared with one-photon excitation fluorescence (OPEF) confocal microscope and wide-field transmission microscope images taken under the same magnification and resolution, TPEF images demonstrate clear advantages in terms of sharpness, signal to noise ratio and contrast. These capabilities provide a working foundation for microendoscopy-based clinical diagnostics of sensorineural hearing loss.

  8. Plasma magnetic field diagnostic using two-photon Doppler-free LIF

    Science.gov (United States)

    Yoon, Young Dae; Bellan, Paul

    2015-11-01

    A detailed description of a new plasma B field diagnostic using Doppler-free two-photon laser-induced fluorescence is presented. The diagnostic is based on a method previously developed in the context of rubidium vapor experiments. Two counter-propagating 393nm diode laser beams are directed into an argon plasma to excite Ar-II ions from 3s2 3p4 4 s4P1 / 2 ⟶ 3s2 3p4 4 p4S3 / 2 ⟶ 3s2 3p4 4 d4P3 / 2 . These levels involve two similar (392.86 and 393.25nm) transition wavelengths, so the two counter-propagating beams effectively cancel out the Doppler effect. The excited ions then decay to the 3s2 3p4 4 p4P1 / 2 level, emitting a 324.98nm line which is to be detected by a photomultiplier tube. The Zeeman splitting -- normally unobservable because of the large Doppler broadening -- of the resultant fluorescence is then to be analyzed, yielding the magnetic field of the particular location. This method is expected to provide a 3-D localized, non-perturbing measurement of magnetic fields. An experimental implementation is currently in progress.

  9. Multi-beam two-photon imaging of fast Ca2+ signals in the Langendorff mouse heart.

    Science.gov (United States)

    Hammer, Karin; Lipp, Peter; Kaestner, Lars

    2014-11-03

    Although the role of calcium (Ca(2+)) in excitation-contraction coupling in the heart can be comprehensively studied at the cellular level, propagation of Ca(2+) signals intercellularly requires tissue-based investigations. To access cells below the epicardium, an optical-sectioning technique is necessary. Multi-photon microscopy allows reliable imaging for penetration to depths of up to 0.5 mm. Here, we provide a protocol that uses multibeam two-photon microscopy for measuring Ca(2+) signals in a Langendorff-perfused mouse heart.

  10. Two-photon Rabi oscillations in a single InxGa1-xAs/GaAs quantum dot

    Science.gov (United States)

    Stufler, S.; Machnikowski, P.; Ester, P.; Bichler, M.; Axt, V. M.; Kuhn, T.; Zrenner, A.

    2006-03-01

    We report on Rabi oscillations of the biexciton in a single InxGa1-xAs/GaAs quantum dot, induced via a coherent two-photon process. In contrast to single-exciton Rabi oscillations the measured population oscillation is not purely sinusoidal in excitation pulse area. The experimental results are well reproduced by a theoretical calculation that relies only on the pulse shape and biexciton binding energy. Dephasing times of the biexciton are similar to those of the single exciton under the applied measurement conditions reaching up to 220ps .

  11. Effect of the coherent cancellation of the two-photon resonance on the generation of vacuum ultraviolet light by two-photon reasonantly enhanced four-wave mixing

    Energy Technology Data Exchange (ETDEWEB)

    Payne, M.G.; Garrett, W.R.; Judish, J.P.; Wunderlich, R.

    1988-11-01

    Many of the most impressive demonstrations of the efficient generation of vacuum ultraviolet (VUV) light have made use of two- photon resonantly enhanced four-wave mixing to generate light at ..omega../sub VUV/ = 2..omega../sub L1/ +- ..omega../sub L2/. The two-photon resonance state is coupled to the ground state both by two photons from the first laser, or by a photon from the second laser and one from the generated VUV beam. We show here that these two coherent pathways destructively interfere once the second laser is made sufficiently intense, thereby leading to an important limiting effect on the achievable conversion efficiency. 4 refs.

  12. Two-photon absorption properties of cationic 1,4-bis(styryl)benzene derivative and its inclusion complexes with cyclodextrins.

    Science.gov (United States)

    Nag, Okhil Kumar; Nayak, Rati Ranjan; Lim, Chang Su; Kim, In Hong; Kyhm, Kwangseuk; Cho, Bong Rae; Woo, Han Young

    2010-07-29

    Two-photon absorption properties of 1,4-bis{4'-[N,N-bis(6''-trimethylammoniumhexyl)amino]styryl}benzene tetrabromide (C1) and its inclusion complexes (ICs) with cyclodextrins (CDs) have been studied. Upon complexation with CDs, the absorption spectra of C1 showed a slight red shift, whereas the emission spectra showed a blue shift with concomitant increase in the fluorescence quantum efficiency. A Stern-Volmer study using K(3)Fe(CN)(6) as a quencher revealed significant reduction in the photoinduced charge transfer quenching, in accord with the IC formation. Comparison of the spectroscopic results reveals that C1 forms increasingly more stable ICs in the order C1/beta-CD < C1/gamma-CD < C1/(3gamma:beta)-CD (gamma-CD/beta-CD 3:1, mole ratio). Moreover, the two-photon action cross section of C1 increased from 200 GM for C1 to 400 GM for C1/beta-CD, 460 GM for C1/gamma-CD, and 650 GM for C1/(3gamma:beta)-CD, respectively. Furthermore, the two-photon microscopy images of HeLa cells stained with C1 emitted strong two-photon excited fluorescence in the plasma membrane. These results provide a useful guideline for the development of efficient two-photon materials for bioimaging applications.

  13. Distribution of quantum information between an atom and two photons

    Energy Technology Data Exchange (ETDEWEB)

    Weber, Bernhard

    2008-11-03

    The construction of networks consisting of optically interconnected processing units is a promising way to scale up quantum information processing systems. To store quantum information, single trapped atoms are among the most proven candidates. By placing them in high finesse optical resonators, a bidirectional information exchange between the atoms and photons becomes possible with, in principle, unit efficiency. Such an interface between stationary and ying qubits constitutes a possible node of a future quantum network. The results presented in this thesis demonstrate the prospects of a quantum interface consisting of a single atom trapped within the mode of a high-finesse optical cavity. In a two-step process, we distribute entanglement between the stored atom and two subsequently emitted single photons. The long atom trapping times achieved in the system together with the high photon collection efficiency of the cavity make the applied protocol in principle deterministic, allowing for the creation of an entangled state at the push of a button. Running the protocol on this quasi-stationary quantum interface, the internal state of the atom is entangled with the polarization state of a single emitted photon. The entanglement is generated by driving a vacuum-stimulated Raman adiabatic passage between states of the coupled atom-cavity system. In a second process, the atomic part of the entangled state is mapped onto a second emitted photon using a similar technique and resulting in a polarization-entangled two-photon state. To verify and characterize the photon-photon entanglement, we measured a violation of a Bell inequality and performed a full quantum state tomography. The results prove the prior atom-photon entanglement and demonstrate a quantum information transfer between the atom and the two emitted photons. This reflects the advantages of a high-finesse cavity as a quantum interface in future quantum networks. (orig.)

  14. Voltage-sensitive rhodol with enhanced two-photon brightness.

    Science.gov (United States)

    Kulkarni, Rishikesh U; Kramer, Daniel J; Pourmandi, Narges; Karbasi, Kaveh; Bateup, Helen S; Miller, Evan W

    2017-03-14

    We have designed, synthesized, and applied a rhodol-based chromophore to a molecular wire-based platform for voltage sensing to achieve fast, sensitive, and bright voltage sensing using two-photon (2P) illumination. Rhodol VoltageFluor-5 (RVF5) is a voltage-sensitive dye with improved 2P cross-section for use in thick tissue or brain samples. RVF5 features a dichlororhodol core with pyrrolidyl substitution at the nitrogen center. In mammalian cells under one-photon (1P) illumination, RVF5 demonstrates high voltage sensitivity (28% ΔF/F per 100 mV) and improved photostability relative to first-generation voltage sensors. This photostability enables multisite optical recordings from neurons lacking tuberous sclerosis complex 1, Tsc1, in a mouse model of genetic epilepsy. Using RVF5, we show that Tsc1 KO neurons exhibit increased activity relative to wild-type neurons and additionally show that the proportion of active neurons in the network increases with the loss of Tsc1. The high photostability and voltage sensitivity of RVF5 is recapitulated under 2P illumination. Finally, the ability to chemically tune the 2P absorption profile through the use of rhodol scaffolds affords the unique opportunity to image neuronal voltage changes in acutely prepared mouse brain slices using 2P illumination. Stimulation of the mouse hippocampus evoked spiking activity that was readily discerned with bath-applied RVF5, demonstrating the utility of RVF5 and molecular wire-based voltage sensors with 2P-optimized fluorophores for imaging voltage in intact brain tissue.

  15. Determining the Quark Charges by One and Two Photon Processes.

    Science.gov (United States)

    Janah, Arjun

    1982-05-01

    Testable predictions are presented, which may be used to decide between the gauge theories of integer and fractionally charged quarks (icq and fcq). Two distinctive features of icq are exploited, namely (a) presence of color non-singlet components in weak and electromagnetic currents and (b) possible liberation of color non-singlet states above a threshold energy. Consequences are sought in lepton-hadron interaction processes, taking into account the known "color-suppression" effect. Single photon/weak-boson processes such as (nu)N (--->) (nu)X distinguish between icq and fcq only above color-threshold. Experimental consequences of color-liberation in the above process are obtained. It is found that the gluon-parton contribution survives color-suppression to produce a significant rise in the structure functions when color-threshold is exceeded. Two-photon processes such as e('+)e('-) (--->) e('+)e('-) + 2 jets distinguish between the two theories even below color threshold. To obtain the icq predictions for this process, one must take into account (a) the (momentum -dependent) color suppression and (b) the added contribution from pair production of charged gluons. This is done, and it is observed that: (i) in icq, the ratio R('(gamma)(gamma)(2 jet)) is not simply a number given by the quark charges; it depends on the gluon mass, on kinematics and on the particular differential cross-section considered; (ii) the deviation of icq cross-sections from the fcq values depends crucially on whether one includes "untagged" events; if this is done, the deviation is large; the charged gluon contribution is mainly responsible for this deviation; the quark contribution is smaller than naively expected. Finally, comparison is made with experimental data on e('+)e('-) (--->) e('+)e('-) + 2 jets. Here, icq is found to be in better agreement than fcq, for a broad range of gluon masses. A suitably modified equivalent photon approximation is employed.

  16. Two-photon path-entangled states in multi-mode waveguides

    CERN Document Server

    Poem, Eilon; Silberberg, Yaron

    2012-01-01

    We experimentally show that two-photon path-entangled states can be coherently manipulated by multi-mode interference in multi-mode waveguides. By measuring the output two-photon spatial correlation function versus the phase of the input state, we show that multi-mode waveguides perform as nearly-ideal multi-port beam splitters at the quantum level, creating a large variety of entangled and separable multi-path two-photon states.

  17. Autofluorescence of routinely hematoxylin and eosin- stained ...

    African Journals Online (AJOL)

    SERVER

    2008-03-04

    Mar 4, 2008 ... histological method, and staining by this dying method is dependant partly upon ... procedure that is specific for it, or for the chemical group to which it belongs ... Hematoxylin was found to have no effect on the inten- ... Hair pulp. -. - Inner root ... Autofluorescence of interstitial tissue makes structures such as ...

  18. Fundus autofluorescence applications in retinal imaging

    Directory of Open Access Journals (Sweden)

    Andrea Gabai

    2015-01-01

    Full Text Available Fundus autofluorescence (FAF is a relatively new imaging technique that can be used to study retinal diseases. It provides information on retinal metabolism and health. Several different pathologies can be detected. Peculiar AF alterations can help the clinician to monitor disease progression and to better understand its pathogenesis. In the present article, we review FAF principles and clinical applications.

  19. Dermal factors influencing measurement of skin autofluorescence

    NARCIS (Netherlands)

    Noordzij, Margaretha J.; Lefrandt, Johan; Graaff, Reindert; Smit, Andries J.

    2011-01-01

    Background: Skin autofluorescence (SAF) is a noninvasive marker of accumulation of advanced glycation end products. It predicts cardiovascular complications and mortality in diabetes and renal failure. We assessed the influence of potential common confounders in SAF measurement, by determining the e

  20. Description of the states of two-photon interference in an optical gating Michelson interferometer

    Science.gov (United States)

    Pongophas, Ekkarat; Sriklin, Watthana; Sinsarp, Asawin; Suwanna, Sujin; Chunwachirasiri, Withoon; Singhsomroje, Wisit

    2016-01-01

    We investigate the interference of two photons in an optical gating Michelson interferometer. The phenomenon is studied using two different representations of photons: the space-time domain and a step-by-step two-photon state evolution. Both representations lead to identical results. The evolution analysis describes the result by the interference of four two-photon traveling states, whereas the space-time domain analysis reveals that the classical interference of the high-intensity light source is identical to two-photon interference in the quantum regime, except for a multiplicative factor of (n2), where n is the number of photons.

  1. Time-reversed two-photon interferometry for phase super-resolution

    CERN Document Server

    Ogawa, Kazuhisa; Kobayashi, Hirokazu; Nakanishi, Toshihiro; Kitano, Masao

    2013-01-01

    We observed two-photon phase super-resolution in an unbalanced Michelson interferometer with classical Gaussian laser pulses. Our work is a time-reversed version of a two-photon interference experiment using an unbalanced Michelson interferometer. A measured interferogram exhibits two-photon phase super-resolution with a high visibility of 97.9% \\pm 0.4%. Its coherence length is about 22 times longer than that of the input laser pulses. It is a classical analogue to the large difference between the one- and two-photon coherence lengths of entangled photon pairs.

  2. Self-assisted optothermal trapping of gold nanorods under two-photon excitation

    Science.gov (United States)

    Chen, Hongtao; Gratton, Enrico; Digman, Michelle A.

    2016-09-01

    We report a self-assisted optothermal trapping and patterning of gold nanorods (GNRs) on glass surfaces with a femtosecond laser. We show that GNRs are not only the trapping targets, but also can enhance the optothermal trapping of other particles. This trapping phenomenon is the net result of thermophoresis and a convective flow caused by localized heating. The heating is due to the conversion of absorbed photons into heat at GNR’s longitudinal surface plasmon resonance (LSPR) wavelength. First, we investigated the optothermal trapping of GNRs at their LSPR wavelength on the glass surface with as low as 0.5 mW laser power. The trapping range was observed to be larger than a typical field of view, e.g. 210 µm  ×  210 µm here. Second, by adjusting the distance between the laser focus and the glass surface, ring patterns of GNRs on the glass surface were obtained. These patterns could be controlled by the laser power and the numerical aperture of the microscope objective. Moreover, we examined the spectral emission of GNRs under different trapping conditions using the spectral phasor approach to reveal the temperature and association status of GNRs. Our study will help understanding manipulation of flows in solution and in biological systems that can be applied in future investigations of GNR-induced heating and flows.

  3. Optically Pumped Atomic Rubidium Lasers: Two-Photon and Exciplex Excitation Mechanisms

    Science.gov (United States)

    2013-06-01

    atomic oxygen”. Physical Review A, 34(1):185–198, 1986. 21. D. Touahri, A. Clairon J. Zondy R. Felder -L. Hilico B. de Beauvoir F. Biraben, O. Acef and F...979, 2004. 24. F. Nez, R. Felder , F. Biraben and Y. Millerioux. “Optical frequency determina- tion of the hyperfine components of the 5S1/2 − 5D3/2

  4. Resonant generation of an electron–positron pair by two photons to excited Landau levels

    Energy Technology Data Exchange (ETDEWEB)

    Diachenko, M. M., E-mail: dyachenko.michail@mail.ru; Novak, O. P.; Kholodov, R. I. [National Academy of Sciences of Ukraine, Institute of Applied Physics (Ukraine)

    2015-11-15

    We consider the resonant generation of an electron–positron pair by two polarized photons to arbitrarily low Landau levels. The resonance occurs when the energy of one photon exceeds the one-photon generation threshold, and the energy of the other photon is multiple to the spacing between the levels. The cross section of the process is determined taking into account the spins of particles. The order of magnitude of the cross section is the highest when the magnetic moments of the particles are oriented along the magnetic field.

  5. Tuning two-photon photoluminescence of gold nanoparticle aggregates with DNA and its application as turn-on photoluminescence probe for DNA sequence detection.

    Science.gov (United States)

    Yuan, Peiyan; Ma, Rizhao; Guan, Zhenping; Gao, Nengyue; Xu, Qing-Hua

    2014-08-13

    Plasmon coupling between noble metal nanoparticles has been known to dramatically enhance linear and nonlinear optical properties of nearby chromophores and metal nanoparticles themselves. The interparticle distance is expected to have significant influence on the coupling strength. Here we have prepared DNA tuned Au nanoparticle assemblies with well controlled separation distances from 2.0 to 12.2 nm to investigate plasmon coupling strength and particle size effects on two-photon photoluminescence (TPPL) enhancement. TPPL intensities of these DNA coupled nanoassemblies were found to increase rapidly as the separation distance decreases. The largest TPPL enhancement factors of 115 and 265 were achieved at the shortest available separation distance of 2.0 nm for 21 and 41 nm Au NPs-dsDNA assemblies, respectively. We have further utilized DNA induced coupling of Au NPs and TPPL enhancement to develop a two-photon sensing scheme for detection of DNA sequences. This TPPL based method displayed high sensitivity with a limit of detection of 2.9 pM and excellent selectivity against ssDNA with mismatched bases. A single mismatch can be easily differentiated at room temperature. Taking the unique advantages of two-photon excitation, this method could be potentially further extended to DNA detection inside cells or even in vivo. These findings can provide important insight for fundamental understanding of plasmon-coupling enhanced TPPL and development of various two-photon excitation based applications.

  6. Test of theoretical models for ultrafast heterogeneous electron transfer with femtosecond two-photon photoemission data

    Indian Academy of Sciences (India)

    Lars Gundlach; Tobias Letzig; Frank Willig

    2009-09-01

    The energy distribution of electrons injected into acceptor states on the surface of TiO2 was measured with femtosecond two-photon photoemission. Shape and relative energetic position of these distribution curves with respect to the corresponding donor states, i.e. of perylene chromophores in the first excited singlet state attached via different bridge-anchor groups to the TiO2 surface, were compared with the predictions of different theoretical models for light-induced ultrafast heterogeneous electron transfer (HET). Gerischer’s early scenario for light-induced HET was considered and two recent explicit calculations, i.e. a fully quantum mechanical analytical model and a time-dependent density functional theory model based on molecular dynamics simulations for the vibrational modes were also considered. Based on the known vibrational structure in the photoionization spectrum of perylene in the gas phase and that measured in the linear absorption spectra of the perylene chromophores anchored on the TiO2 surface the energy distribution curves for the injected electrons were fitted assuming the excitation of the dominant 0.17 eV vibrational mode in the ionized perylene chromophore leading to a corresponding Franck-Condon dictated progression in the energy distribution curves. Each individual peak was fitted with a Voigt profile where the Lorentzian contribution was taken from the time-resolved HET data and the Gaussian contribution attributed to inhomogeneous broadening. The measured room temperature energy distribution curves for the injected electrons are explained with the fully quantum mechanical model for light-induced HET with the high energy, 0.17 eV, skeletal stretching mode excited in the ionized perylene chromophore. The corresponding energy distribution of the injected electrons is fully accommodated in acceptor states on the TiO2 surface fulfilling the wide band limit.

  7. Giant Two-Photon Absorption Coefficient and Frequency Up-Converted Luminescence in Monolayer MoS2

    CERN Document Server

    Li, Yuanxin; Zhang, Saifeng; Zhang, Xiaoyan; Feng, Yanyan; Wang, Kangpeng; Zhang, Long; Wang, Jun

    2015-01-01

    Strong two-photon absorption (TPA) in monolayer MoS2 is demonstrated in contrast to saturable absorption (SA) in multilayer MoS2 under the excitation of femtosecond laser pulses in the near infrared region. MoS2 in the forms of monolayer single crystal and multilayer triangular islands are grown on either quartz or SiO2/Si by employing the seeding method through chemistry vapor deposition. The nonlinear transmission measurements reveal that monolayer MoS2 possesses a giant nonsaturation TPA coefficient, larger than that of conventional semiconductors. As a result of TPA, two-photon pumped frequency up-converted luminescence is observed directly in the monolayer MoS2. For the multilayer MoS2, the SA response is demonstrated with the ratio of the excited-state absorption cross section to ground-state cross section of 0.18. In addition, the laser damage threshold of the monolayer MoS2 is 97 GW/cm2, larger than that of the multilayer MoS2 of 78 GW/cm2.

  8. Two-photon luminescence contrast by tip-sample coupling in femtosecond near-field optical microscopy

    Science.gov (United States)

    Horneber, Anke; Wackenhut, Frank; Braun, Kai; Wang, Xiao; Wang, Jiyong; Zhang, Dai; Meixner, Alfred J.

    2017-01-01

    We investigate the role of tip-sample interaction in nonlinear optical scanning near-field microscopy. The experiment was performed by tightly focusing femtosecond laser pulses onto a sharp gold tip that was positioned in close proximity to the surface of a sample with gold nanostructures on a Si-substrate by shear force feedback. The nonlinear optical signal consists of two-photon photoluminescence and second harmonic signal from the gold tip and the gold nanostructures. These signals can be used to characterize different coupling parameters such as geometry, material and width of the tip-sample gap and enable to reveal the mechanism responsible for the image contrast. Under the excitation with 776-nm and 110-fs laser pulses nonlinear imaging is almost background free and yields super resolution showing features with dimensions significantly below the diffraction limit with a signal intensity following quadratic excitation power law.

  9. High-accuracy reference standards for two-photon absorption in the 680-1050 nm wavelength range.

    Science.gov (United States)

    de Reguardati, Sophie; Pahapill, Juri; Mikhailov, Alexander; Stepanenko, Yuriy; Rebane, Aleksander

    2016-04-18

    Degenerate two-photon absorption (2PA) of a series of organic fluorophores is measured using femtosecond fluorescence excitation method in the wavelength range, λ2PA = 680-1050 nm, and ~100 MHz pulse repetition rate. The function of relative 2PA spectral shape is obtained with estimated accuracy 5%, and the absolute 2PA cross section is measured at selected wavelengths with the accuracy 8%. Significant improvement of the accuracy is achieved by means of rigorous evaluation of the quadratic dependence of the fluorescence signal on the incident photon flux in the whole wavelength range, by comparing results obtained from two independent experiments, as well as due to meticulous evaluation of critical experimental parameters, including the excitation spatial- and temporal pulse shape, laser power and sample geometry. Application of the reference standards in nonlinear transmittance measurements is discussed.

  10. Enhancement of Squeezing in Two-Photon Jaynes-Cummings Model with Atomic Measurement

    Institute of Scientific and Technical Information of China (English)

    YE Sai-Yun

    2006-01-01

    We investigate the squeezing properties of the cavity field in the degenerate two-photon Jaynes-Cummings model. Compared with the one-photon Jaynes-Cummings model, the squeezing is more pronounced in the case of two-photon Jaynes-Cummings model under certain conditions.

  11. Event-by-event simulation of nonclassical effects in two-photon interference experiments

    NARCIS (Netherlands)

    Michielsen, K.; Jin, F.; Delina, M.; Raedt, H. De

    2012-01-01

    A corpuscular simulation model for second-order intensity interference phenomena is discussed. It is shown that both the visibility V = 1/2 predicted for two-photon interference experiments with two independent sources and the visibility V = 1 predicted for two-photon interference experiments with a

  12. Integrated single- and two-photon light sheet microscopy using accelerating beams

    DEFF Research Database (Denmark)

    Piksarv, Peeter; Marti, Dominik; Le, Tuan

    2017-01-01

    We demonstrate the first light sheet microscope using propagation invariant, accelerating Airy beams that operates both in single- and two-photon modes. The use of the Airy beam permits us to develop an ultra compact, high resolution light sheet system without beam scanning. In two-photon mode, a...

  13. Two-photon fluorescence probes for imaging of mitochondria and lysosomes.

    Science.gov (United States)

    Yang, Wanggui; Chan, Pui Shan; Chan, Miu Shan; Li, King Fai; Lo, Pik Kwan; Mak, Nai Ki; Cheah, Kok Wai; Wong, Man Shing

    2013-04-28

    Novel biocompatible cyanines show not only a very large two-photon cross-section of up to 5130 GM at 910 nm in aqueous medium for high-contrast and -brightness two-photon fluorescence live cell imaging but also highly selective subcellular localization properties including localization of mitochondria and lysosomes.

  14. Effect of morphology and solvent on two-photon absorption of nano zinc oxide

    Energy Technology Data Exchange (ETDEWEB)

    Kavitha, M.K. [Department of Chemistry, Indian Institute of Space Science and Technology, Valiamala, Thiruvananthapuram 695547, Kerala (India); Haripadmam, P.C.; Gopinath, Pramod; Krishnan, Bindu [Department of Physics, Indian Institute of Space Science and Technology, Valiamala, Thiruvananthapuram 695547, Kerala (India); John, Honey, E-mail: honey@iist.ac.in [Department of Chemistry, Indian Institute of Space Science and Technology, Valiamala, Thiruvananthapuram 695547, Kerala (India)

    2013-05-15

    Highlights: ► ZnO nanospheres and triangular structures synthesis by novel precipitation technique. ► The effect of precursor concentration on the size and shape of nano ZnO. ► Open aperture Z-scan measurements of the ZnO nanoparticle dispersions. ► Nanospheres exhibit higher two photon absorption coefficient than triangular nanostructures. ► Nanospheres dispersed in water exhibit higher two photon absorption coefficient than its dispersion in 2-propanol. - Abstract: In this paper, we report the effect of morphology and solvent on the two-photon absorption of nano zinc oxide. Zinc oxide nanoparticles in two different morphologies like nanospheres and triangular nanostructures are synthesized by novel precipitation technique and their two-photon absorption coefficient is measured using open aperture Z-scan technique. Experimental results show that the zinc oxide nanospheres exhibit higher two-photon absorption coefficient than the zinc oxide triangular nanostructures. The zinc oxide nanospheres dispersed in water exhibit higher two-photon absorption coefficient than that of its dispersion in 2-propanol. The zinc oxide nanospheres dispersed in water shows a decrease in two-photon absorption coefficient with an increase in on-axis irradiance. The result confirms the dependence of shape and solvent on the two-photon absorption of nano zinc oxide.

  15. Production of e, $\\mu$ and $\\tau$ Pairs in Untagged Two-Photon Collisions at LEP

    CERN Document Server

    Acciarri, M; Aguilar-Benítez, M; Ahlen, S P; Alcaraz, J; Alemanni, G; Allaby, James V; Aloisio, A; Alverson, G; Alviggi, M G; Ambrosi, G; Anderhub, H; Andreev, V P; Angelescu, T; Anselmo, F; Arefev, A; Azemoon, T; Aziz, T; Bagnaia, P; Baksay, L; Ball, R C; Banerjee, S; Banerjee, Sw; Banicz, K; Barczyk, A; Barillère, R; Barone, L; Bartalini, P; Baschirotto, A; Basile, M; Battiston, R; Bay, A; Becattini, F; Becker, U; Behner, F; Berdugo, J; Berges, P; Bertucci, B; Betev, B L; Bhattacharya, S; Biasini, M; Biland, A; Bilei, G M; Blaising, J J; Blyth, S C; Bobbink, Gerjan J; Böck, R K; Böhm, A; Boldizsar, L; Borgia, B; Boucham, A; Bourilkov, D; Bourquin, Maurice; Boutigny, D; Braccini, S; Branson, J G; Brigljevic, V; Brock, I C; Buffini, A; Buijs, A; Burger, J D; Burger, W J; Busenitz, J K; Cai, X D; Campanelli, M; Capell, M; Cara Romeo, G; Carlino, G; Cartacci, A M; Casaus, J; Castellini, G; Cavallari, F; Cavallo, N; Cecchi, C; Cerrada-Canales, M; Cesaroni, F; Chamizo-Llatas, M; Chang, Y H; Chaturvedi, U K; Chekanov, S V; Chemarin, M; Chen, A; Chen, G; Chen, G M; Chen, H F; Chen, H S; Chen, M; Chiefari, G; Chien, C Y; Cifarelli, Luisa; Cindolo, F; Civinini, C; Clare, I; Clare, R; Cohn, H O; Coignet, G; Colijn, A P; Colino, N; Commichau, V; Costantini, S; Cotorobai, F; de la Cruz, B; Csilling, Akos; Dai, T S; D'Alessandro, R; De Asmundis, R; Degré, A; Deiters, K; Denes, P; De Notaristefani, F; DiBitonto, Daryl; Diemoz, M; Van Dierendonck, D N; Di Lodovico, F; Dionisi, C; Dittmar, Michael; Dominguez, A; Doria, A; Dorne, I; Dova, M T; Drago, E; Duchesneau, D; Duinker, P; Durán, I; Dutta, S; Easo, S; Efremenko, Yu V; El-Mamouni, H; Engler, A; Eppling, F J; Erné, F C; Ernenwein, J P; Extermann, Pierre; Fabre, M; Faccini, R; Falciano, S; Favara, A; Fay, J; Fedin, O; Felcini, Marta; Fenyi, B; Ferguson, T; Ferroni, F; Fesefeldt, H S; Fiandrini, E; Field, J H; Filthaut, Frank; Fisher, P H; Fisk, I; Forconi, G; Fredj, L; Freudenreich, Klaus; Furetta, C; Galaktionov, Yu; Ganguli, S N; 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    1997-01-01

    The two-photon collision reaction e+e- --> e+e-l+l- has been studied at root(s) ~ 91 GeV using the L3 detector at LEP for l = e, muon , tau. We have analysed untagged configurations where the two photons are quasi-real. Good agreement is found between our measurements and the order alpha**4 QED expectation.

  16. Temporal dynamics of two-photon-pumped amplified spontaneous emission in slab organic crystals

    NARCIS (Netherlands)

    Fang, Hong-Hua; Chen, Qi-Dai; Ding, Ran; Yang, Jie; Ma, Yu-Guang; Wang, Hai-Yu; Gao, Bing-Rong; Feng, Jing; Sun, Hong-Bo; Fang, Honghua

    2010-01-01

    We have studied the ultrafast dynamics of two-photon-pumped amplified spontaneous emission (ASE) from a single crystal by the time-resolved fluorescence upconversion technique. With the increase of two-photon pump intensities, the emission decay time is dramatically shortened by 30 times (from 3 ns

  17. Two-Photon Interference with the Type Ⅱ Spontaneous Parametric Down-Conversion

    Institute of Scientific and Technical Information of China (English)

    江云坤; 史保森; 李剑; 段开敏; 范晓锋; 郭光灿

    2001-01-01

    The two-photon polarized entangled state is generated from the type Ⅱ spontaneous parametric down-conversion pumped by a femtosecond pulse. The two-photon interference is observed in the Hong-Ou-Mandel interferometer. The high visibility of the interference is restored with narrow band interference filters placed in front of the detectors.

  18. Search for two Higgs bosons in final states containing two photons and two bottom quarks

    Energy Technology Data Exchange (ETDEWEB)

    Khachatryan, Vardan; et al.

    2016-03-22

    A search is presented for the production of two Higgs bosons in final states containing two photons and two bottom quarks. Both resonant and nonresonant hypotheses are investigated. The analyzed data correspond to an integrated luminosity of 19.7 inverse femtobarns of proton-proton collisions at sqrt(s) = 8 TeV collected with the CMS detector. Good agreement is observed between data and predictions of the standard model (SM). Upper limits are set at 95% confidence level on the production cross section of new particles and compared to the prediction for the existence of a warped extra dimension. When the decay to two Higgs bosons is kinematically allowed, assuming a mass scale LambdaR = 1 TeV for the model, the data exclude a radion scalar at masses below 980 GeV. The first Kaluza-Klein excitation mode of the graviton in Randall-Sundrum models is excluded for masses between 325 and 450 GeV. Limits set on nonresonant production constrain the parameter space for anomalous Higgs boson couplings.

  19. Zn2+ responsive two-photon fluorescent probes based on branch structure: a computational investigation

    Science.gov (United States)

    Huang, Shuang; Yang, Bao-Zhu; Guo, Jing-Fu; Ren, Ai-Min

    2015-03-01

    A series of zinc ion fluorescent probes on the basis of multi-branched ligands were investigated in theory. The three-branched ligand TPPA (N,N,N‧,N‧-tetraphenyl-p-phenylenediamine) has better three-dimensional spatial localisation, which can detect zinc at the parts per million level. The complex coordinated with Zn2+ can show a significant improvement in two-photon absorption (TPA) cross-section in the near-infrared (NIR) excitation region. The calculated results reveal that the stability and sensitivity of Zn2+ complexes will be enhanced by increasing the number of branches. The selectivity of double phenyl-p-phenylenediamine (DPPA) ligand to Zn2+ will be better compared to Cd2+. With regard to the studied ligands single phenyl-p-phenylenediamine (SPPA), two connected single phenyl-p-phenylenediamine (2CSPPA), DPPA and TPPA, λEMmax shows a red-shift and ƒEM gets stronger upon the addition of Zn2+. Most of the molecules exhibit TPA peaks in the NIR region. The theoretical investigations demonstrate that DPPA-Zn2+ shows good TPA activity at a telecommunication wavelength.

  20. Multispot two-photon imaging of mice heart tissue detecting calcium waves

    Science.gov (United States)

    de Mauro, C.; Cecchetti, C. A.; Alfieri, D.; Borile, G.; Mongillo, M.; Pavone, F. S.

    2012-06-01

    High rate, full field image acquisition in multiphoton imaging is achievable by parallelization of the excitation and of the detection paths. Via a Diffractive Optical Elements (DOEs) which splits a pulsed laser, and a spatial resolved descanned detection path, a new approach to microscopy has been developed. By exploiting the three operating mode, single beam, 16 beamlets or 64 beamlets, the best experimental conditions can be found by adapting the power per beamlet. This Multiphoton Multispot system (MCube) has been characterized in thick tissue samples, and subsequently used for the first time for Ca2+ imaging of acute heart slices. A test sample with fixed mice heart slices with embedded sub-resolution fluorescent beads has been used to test the capability of optical axial resolution up to ~200 microns in depth. Radial and axial resolutions of 0.6 microns and 3 microns have been respectively obtained with a 40X water immersion objective, getting close to the theoretical limit. Then images of heart slices cardiomyocites, loaded with Fluo4-AM have been acquired. The formation of Ca2+ waves during electrostimulated beating has been observed, and the possibility of easily acquire full frame images at 15 Hz (16 beamlets) has been demonstrated, towards the in vivo study of time resolved cellular dynamics and arrhythmia trigger mechanisms in particular. A very high speed two-photon Random Access system for in vivo electrophysiological studies, towards the correlation of voltage and calcium signals in arrhythmia phenomena, is now under developing at Light4tech.