WorldWideScience

Sample records for scanless two-photon imaging

  1. 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.

  2. A simple scanless two-photon fluorescence microscope using selective plane illumination.

    Science.gov (United States)

    Palero, Jonathan; Santos, Susana I C O; Artigas, David; Loza-Alvarez, Pablo

    2010-04-12

    We demonstrate a simple scanless two-photon (2p) excited fluorescence microscope based on selective plane illumination microscopy (SPIM). Optical sectioning capability is presented and depth-resolved imaging of cameleon protein in C. elegans pharyngeal muscle is implemented.

  3. Denoising two-photon calcium imaging data.

    Directory of Open Access Journals (Sweden)

    Wasim Q Malik

    Full Text Available 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.

  4. 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.

  5. 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.

  6. Two-photon imaging with diffractive optical elements

    Directory of Open Access Journals (Sweden)

    Brendon O Watson

    2009-07-01

    Full Text Available Two-photon imaging has become a useful tool for optical monitoring of neural circuits, but it requires high laser power and serial scanning of each pixel in a sample. This results in slow imaging rates, limiting the measurements of fast signals such as neuronal activity. To improve the speed and signal-to-noise ratio of two-photon imaging, we introduce a simple modification of a two-photon microscope, using a diffractive optical element (DOE which splits the laser beam into several beamlets that can simultaneously scan the sample. We demonstrate the advantages of DOE scanning by enhancing the speed and sensitivity of two-photon calcium imaging of action potentials in neurons from neocortical brain slices. DOE scanning can easily improve the detection of time-varying signals in two-photon and other non-linear microscopies.

  7. Two-photon excitation based photochemistry and neural imaging

    Science.gov (United States)

    Hatch, Kevin Andrew

    Two-photon microscopy is a fluorescence imaging technique which provides distinct advantages in three-dimensional cellular and molecular imaging. The benefits of this technology may extend beyond imaging capabilities through exploitation of the quantum processes responsible for fluorescent events. This study utilized a two-photon microscope to investigate a synthetic photoreactive collagen peptidomimetic, which may serve as a potential material for tissue engineering using the techniques of two-photon photolysis and two-photon polymerization. The combination of these techniques could potentially be used to produce a scaffold for the vascularization of engineered three-dimensional tissues in vitro to address the current limitations of tissue engineering. Additionally, two-photon microscopy was used to observe the effects of the application of the neurotransmitter dopamine to the mushroom body neural structures of Drosophila melanogaster to investigate dopamine's connection to cognitive degeneration.

  8. Two-Photon Imaging of the Mouse Eye

    OpenAIRE

    Johnson, Andrew W.; Ammar, David A.; Kahook, Malik Y.

    2011-01-01

    The authors used two-photon microscopy to image structures of the conventional aqueous humor outflow pathway in an intact mouse eye. This imaging method may be useful for following the progress of animal models of glaucoma in a noninvasive manner.

  9. Two-photon optical microscopy imaging of endothelial keratoplasty grafts.

    Science.gov (United States)

    Lombardo, Marco; Parekh, Mohit; Serrao, Sebastiano; Ruzza, Alessandro; Ferrari, Stefano; Lombardo, Giuseppe

    2017-03-01

    To investigate the microstructure of endothelial keratoplasty grafts using two-photon optical microscopy. Six endothelial keratoplasty grafts obtained from human donor corneoscleral tissues and prepared by submerged hydrodissection technique were imaged by two-photon optical microscopy. In each graft, two liquid bubbles were created in order to investigate the presence of a conserved cleavage plane regardless of the volume of posterior stroma that remained attached to Descemet's membrane (DM); the first bubble (bubble A) was generated under DM and the second bubble (bubble B) injection was done in order to obtain a layer of deep stroma that kept the two bubbles separated. Six human donor corneoscleral tissues were used as controls. Second harmonic generation and two-photon emitted fluorescence signals were collected from each specimen. Dissection of stroma occurred along the posterior collagen lamellae at variable distance from DM, which ranged between 3 and 16 μm in bubble A and between 23 and 41 μm in bubble B. The residual stroma included, anteriorly, bands of collagen lamellae, and thin bundles of stromal collagen fibrils, posteriorly, which were tightly intertwining with the underlying DM. There was no anatomically distinct plane of separation between these pre-Descemetic stromal collagen bundles and the overlying collagen lamellae with this hydrodissection technique. Two-photon optical microscopy provided label-free high-resolution imaging of endothelial keratoplasty grafts, showing that the most posterior stroma changes organization at approximately 10 μm above the DM. The pre-Descemetic stromal collagen fibrils form an intertwined complex with DM, which cannot be separated using hydrodissection.

  10. 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.

  11. 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.

  12. 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

  13. Multidimensional two-photon imaging of diseased skin

    Science.gov (United States)

    Cicchi, R.; Sestini, S.; De Giorgi, V.; Massi, D.; Lotti, T.; Pavone, F. S.

    2008-02-01

    We used combined two photon intrinsic fluorescence (TPE), second harmonic generation microscopy (SHG), fluorescence lifetime imaging microscopy (FLIM), and multispectral two photon emission detection (MTPE) to investigate different kinds of human cutaneous ex-vivo skin lesions. Morphological and spectroscopic analyses allowed to characterize both healthy and pathological skin samples, including tumors, as well as to discriminate between healthy and diseased tissue, in a good agreement with common routine histology. In particular, we examined tissue samples from normal and pathological scar tissue (keloid), and skin tumors, including basal cell carcinoma (BCC) and malignant melanoma (MM). By using combined TPE-SHG microscopy we investigated morphological features of different skin regions, as BCC, tumor-stroma interface, healthy dermis, fibroblastic proliferation, and keloids. The SHG to autofluorescence aging index of dermis (SAAID) score was used to characterize each region, finding differences between BCC, healthy skin, tumor-stroma interface, keloids, and fibroblastic proliferation. Further comparative analysis of healthy skin and neoplastic samples was performed using FLIM. In particular, BCC showed a blue-shifted fluorescence emission, a higher absorption at 800 nm excitation wavelength, and a slightly longer mean fluorescence lifetime. MM showed a lifetime distribution similar to the corresponding melanocytic nevus (MN) lifetime distribution for the slow lifetime component, and different for the fast lifetime component.

  14. Two-photon intravital imaging of thrombus development

    Science.gov (United States)

    Kamocka, Malgorzata M.; Mu, Jian; Liu, Xiaomin; Chen, Nan; Zollman, Amy; Sturonas-Brown, Barbara; Dunn, Kenneth; Xu, Zhiliang; Chen, Danny Z.; Alber, Mark S.; Rosen, Elliot D.

    2010-01-01

    Thrombus development in mouse mesenteric vessels following laser-induced injury was monitored by high-resolution, near-real-time, two-photon, intravital microscopy. In addition to the use of fluorescently tagged fibrin(ogen) and platelets, plasma was labeled with fluorescently tagged dextran. Because blood cells exclude the dextran in the single plane, blood cells appear as black silhouettes. Thus, in addition to monitoring the accumulation of platelets and fibrin in the thrombus, the protocol detects the movement and incorporation of unlabeled cells in and around it. The developing thrombus perturbs the blood flow near the thrombus surface, which affects the incorporation of platelets and blood cells into the structure. The hemodynamic effects and incorporation of blood cells lead to the development of thrombi with heterogeneous domain structures. Additionally, image processing algorithms and simulations were used to quantify structural features of developing thrombi. This analysis suggests a novel mechanism to stop the growth of developing thrombus.

  15. Porphyrin- or phthalocyanine-bridged silsesquioxane nanoparticles for two-photon photodynamic therapy or photoacoustic imaging.

    Science.gov (United States)

    Mauriello-Jimenez, Chiara; Henry, Maxime; Aggad, Dina; Raehm, Laurence; Cattoën, Xavier; Wong Chi Man, Michel; Charnay, Clarence; Alpugan, Serkan; Ahsen, Vefa; Tarakci, Deniz Kutlu; Maillard, Philippe; Maynadier, Marie; Garcia, Marcel; Dumoulin, Fabienne; Gary-Bobo, Magali; Coll, Jean-Luc; Josserand, Véronique; Durand, Jean-Olivier

    2017-11-09

    Porphyrin- or phthalocyanine-bridged silsesquioxane nanoparticles (BSPOR and BSPHT) were prepared. Their endocytosis in MCF-7 cancer cells was shown with two-photon excited fluorescence (TPEF) imaging. With two-photon excited photodynamic therapy (TPE-PDT), BSPOR was more phototoxic than BSPHT, which in contrast displayed a very high signal for photoacoustic imaging in mice.

  16. Gold nanorods as dual photo-sensitizing and imaging agents for two-photon photodynamic therapy

    Science.gov (United States)

    Zhao, Tingting; Shen, Xiaoqin; Li, Lin; Guan, Zhenping; Gao, Nengyue; Yuan, Peiyan; Yao, Shao Q.; Xu, Qing-Hua; Xu, Guo Qin

    2012-11-01

    Gold nanorods with three different aspect ratios were prepared and their dual capabilities for two-photon imaging and two-photon photodynamic therapy have been demonstrated. These gold nanorods exhibit large two-photon absorption action cross-sections, about two orders of magnitude larger than small organic molecules, which makes them suitable for two-photon imaging. They can also effectively generate singlet oxygen under two-photon excitation, significantly higher than traditional photosensitizers such as Rose Bengal and Indocyanine Green. Such high singlet oxygen generation capability under two-photon excitation was ascribed to their large two-photon absorption cross-sections. Polyvinylpyrrolidone (PVP) coated gold nanorods displayed excellent biocompatibility and high cellular uptake efficiency. The two-photon photodynamic therapy effect and two-photon fluorescence imaging properties of PVP coated gold nanorods have been successfully demonstrated on HeLa cells in vitro using fluorescence microscopy and indirect XTT assay method. These gold nanorods thus hold great promise for imaging guided two-photon photodynamic therapy for the treatment of various malignant tumors.Gold nanorods with three different aspect ratios were prepared and their dual capabilities for two-photon imaging and two-photon photodynamic therapy have been demonstrated. These gold nanorods exhibit large two-photon absorption action cross-sections, about two orders of magnitude larger than small organic molecules, which makes them suitable for two-photon imaging. They can also effectively generate singlet oxygen under two-photon excitation, significantly higher than traditional photosensitizers such as Rose Bengal and Indocyanine Green. Such high singlet oxygen generation capability under two-photon excitation was ascribed to their large two-photon absorption cross-sections. Polyvinylpyrrolidone (PVP) coated gold nanorods displayed excellent biocompatibility and high cellular uptake efficiency

  17. Two-photon fluorescence imaging of intracellular hydrogen peroxide with chemoselective fluorescent probes

    OpenAIRE

    Guo, Hengchang; Aleyasin, Hossein; Howard, Scott S.; Dickinson, Bryan C.; Lin, Vivian S.; Haskew-Layton, Renee E.; Xu, Chris; Chen, Yu; Ratan, Rajiv R.

    2013-01-01

    Abstract. We present the application of two-photon fluorescence (TPF) imaging to monitor intracellular hydrogen peroxide (H2O2) production in brain cells. For selective imaging of H2O2 over other reactive oxygen species, we employed small-molecule fluorescent probes that utilize a chemoselective boronate deprotection mechanism. Peroxyfluor-6 acetoxymethyl ester detects global cellular H2O2 and mitochondria peroxy yellow 1 detects mitochondrial H2O2. Two-photon absorption cross sections for th...

  18. Images of photoreceptors in living primate eyes using adaptive optics two-photon ophthalmoscopy

    Science.gov (United States)

    Hunter, Jennifer J.; Masella, Benjamin; Dubra, Alfredo; Sharma, Robin; Yin, Lu; Merigan, William H.; Palczewska, Grazyna; Palczewski, Krzysztof; Williams, David R.

    2011-01-01

    In vivo two-photon imaging through the pupil of the primate eye has the potential to become a useful tool for functional imaging of the retina. Two-photon excited fluorescence images of the macaque cone mosaic were obtained using a fluorescence adaptive optics scanning laser ophthalmoscope, overcoming the challenges of a low numerical aperture, imperfect optics of the eye, high required light levels, and eye motion. Although the specific fluorophores are as yet unknown, strong in vivo intrinsic fluorescence allowed images of the cone mosaic. Imaging intact ex vivo retina revealed that the strongest two-photon excited fluorescence signal comes from the cone inner segments. The fluorescence response increased following light stimulation, which could provide a functional measure of the effects of light on photoreceptors. PMID:21326644

  19. [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.

  20. 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.

  1. Multifunctional hybrid nanoparticles for two-photon fluorescence imaging and photodynamic therapy

    Science.gov (United States)

    Baldeck, Patrice L.; Maurin, Mathieu; Philipot, Cecile; Zaiba, Soraya; Gallavardin, Thibault; Maury, Olivier; Andraud, Chantal; Dubois, Fabien; Ibanez, Alain; Lerouge, Frédéric; Parola, Stéphane; Stephan, Olivier; van Der Sanden, Baudwin

    2011-02-01

    We review our work on several strategies to elaborate multifunctional nanoparticules for two-photon imaging or/and photodynamic therapy. Our first strategy is based on the incorporation of two-photon hydrophobic fluorophors in bio-compatible pluronic micelles using the mini-emulsion technique. Our second strategy is based on fluorescent organic nanocrystal grown in silicate spheres. These core-shell hybrid nanoparticles are obtained by a spray-drying process from sol-gel solutions. Our third strategy consists in the encapsulation of hydrophilic molecules in the water core of gold nanospheres. They are obtained by a stabilized emulsion in biphasic liquid-liquid medium without surfactant.

  2. Two-photon imaging using adaptive phase compensated ultrashort laser pulses.

    Science.gov (United States)

    Xi, Peng; Andegeko, Yair; Pestov, Dmitry; Lovozoy, Vadim V; Dantus, Marcos

    2009-01-01

    An adaptive pulse shaper controlled by multiphoton intrapulse interference phase scanning (MIIPS) was used, together with a prism-pair, to measure and cancel high-order phase distortions introduced by a high-numerical-aperture objective and other dispersive elements of a two-photon laser-scanning microscope. The delivery of broad-bandwidth (approximately 100 nm), sub-12-fs pulses was confirmed by interferometric autocorrelation measurements at the focal plane. A comparison of two-photon imaging with transform-limited and second-order-dispersion compensated laser pulses of the same energy showed a 6-to-11-fold improvement in the two-photon excitation fluorescence signal when applied to cells and tissue, and up to a 19-fold improvement in the second harmonic generation signal from a rat tendon specimen.

  3. 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

  4. Video-rate two-photon excited fluorescence lifetime imaging system with interleaved digitization.

    Science.gov (United States)

    Dow, Ximeng Y; Sullivan, Shane Z; Muir, Ryan D; Simpson, Garth J

    2015-07-15

    A fast (up to video rate) two-photon excited fluorescence lifetime imaging system based on interleaved digitization is demonstrated. The system is compatible with existing beam-scanning microscopes with minor electronics and software modification. Proof-of-concept demonstrations were performed using laser dyes and biological tissue.

  5. Gold Core Mesoporous Organosilica Shell Degradable Nanoparticles for Two-Photon Imaging and Gemcitabine Monophosphate Delivery

    KAUST Repository

    Rhamani, Saher

    2017-09-12

    The synthesis of gold core degradable mesoporous organosilica shell nanoparticles is described. The nanopaticles were very efficient for two-photon luminescence imaging of cancer cells and for in vitro gemcitabine monophosphate delivery, allowing promising theranostic applications in the nanomedicine field.

  6. Two-photon imaging of calcium in virally transfected striate cortical neurons of behaving monkey.

    Directory of Open Access Journals (Sweden)

    Barbara Heider

    2010-11-01

    Full Text Available Two-photon scanning microscopy has advanced our understanding of neural signaling in non-mammalian species and mammals. Various developments are needed to perform two-photon scanning microscopy over prolonged periods in non-human primates performing a behavioral task. In striate cortex in two macaque monkeys, cortical neurons were transfected with a genetically encoded fluorescent calcium sensor, memTNXL, using AAV1 as a viral vector. By constructing an extremely rigid and stable apparatus holding both the two-photon scanning microscope and the monkey's head, single neurons were imaged at high magnification for prolonged periods with minimal motion artifacts for up to ten months. Structural images of single neurons were obtained at high magnification. Changes in calcium during visual stimulation were measured as the monkeys performed a fixation task. Overall, functional responses and orientation tuning curves were obtained in 18.8% of the 234 labeled and imaged neurons. This demonstrated that the two-photon scanning microscopy can be successfully obtained in behaving primates.

  7. Two-photon imaging of calcium in virally transfected striate cortical neurons of behaving monkey.

    Science.gov (United States)

    Heider, Barbara; Nathanson, Jason L; Isacoff, Ehud Y; Callaway, Edward M; Siegel, Ralph M

    2010-11-04

    Two-photon scanning microscopy has advanced our understanding of neural signaling in non-mammalian species and mammals. Various developments are needed to perform two-photon scanning microscopy over prolonged periods in non-human primates performing a behavioral task. In striate cortex in two macaque monkeys, cortical neurons were transfected with a genetically encoded fluorescent calcium sensor, memTNXL, using AAV1 as a viral vector. By constructing an extremely rigid and stable apparatus holding both the two-photon scanning microscope and the monkey's head, single neurons were imaged at high magnification for prolonged periods with minimal motion artifacts for up to ten months. Structural images of single neurons were obtained at high magnification. Changes in calcium during visual stimulation were measured as the monkeys performed a fixation task. Overall, functional responses and orientation tuning curves were obtained in 18.8% of the 234 labeled and imaged neurons. This demonstrated that the two-photon scanning microscopy can be successfully obtained in behaving primates.

  8. Two-Photon Photosensitizer-Polymer Conjugates for Combined Cancer Cell Death Induction and Two-Photon Fluorescence Imaging: Structure/Photodynamic Therapy Efficiency Relationship.

    Science.gov (United States)

    Cepraga, Cristina; Marotte, Sophie; Ben Daoud, Edna; Favier, Arnaud; Lanoë, Pierre-Henri; Monnereau, Cyrille; Baldeck, Patrice; Andraud, Chantal; Marvel, Jacqueline; Charreyre, Marie-Thérèse; Leverrier, Yann

    2017-11-10

    One of the challenges of photodynamic therapy is to increase the penetration depth of light irradiation in the tumor tissues. Although two-photon excitation strategies have been developed, the two-photon absorption cross sections of clinically used photosensitizers are generally low (below 300 GM). Besides, photosensitizers with high cross section values are often non-water-soluble. In this research work, a whole family of photosensitizer-polymer conjugates was synthesized via the covalent binding of a photosensitizer with a relatively high cross section along a biocompatible copolymer chain. The resulting photosensitizer-polymer conjugates were water-soluble and could be imaged in cellulo by two-photon microscopy thanks to their high two-photon absorption cross sections (up to 2600 GM in water, in the NIR range). In order to explore the structure/photodynamic activity relationship of such macromolecular photosensitizers, the influence of the polymer size, photosensitizer density, and presence of charges along the polymer backbone was investigated (neutral, anionic, cationic, and zwitterionic conjugates were compared). The macromolecular photosensitizers were not cytotoxic in the absence of light irradiation. Their kinetics of cellular uptake in the B16-F10 melanoma cell line were followed by flow cytometry over 24 h. The efficiency of cell death upon photoactivation was found to be highly correlated to the cellular uptake in turn correlated to the global charge of the macromolecular photosensitizer which appeared as the determining structural parameter.

  9. Two-photon excited fluorescence lifetime imaging microscopy for FRET study on protein interactions

    Science.gov (United States)

    Qu, Junle; Lin, Ziyang; Liu, Lixin; Guo, Xuan; Chen, Danni; Niu, Hanben

    2005-01-01

    Two-photon excited fluorescence lifetime imaging (2P-FLIM) provides a more direct and precise approach to fluorescence resonance energy transfer (FRET), which allows studying the dynamic behavior of protein-protein interactions in living cells. In this paper, we describe the combination of a Leica TCS SP2 laser scanning microscope and a time-correlated single photon counting (TCSPC) lifetime imaging module developed by Becker & Hickl for two-photon excited fluorescence lifetime imaging. This 2P-FLIM system was used for FRET study on the interaction of heat shock protein hsp27 with p38 MAP kinase in the single living cell. Results show that the reduction in donor (CFP) lifetime in the presence of acceptor (YFP) reveals interactions between the two proteins.

  10. 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.

  11. 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.

  12. Optimal repetition rate and pulse duration studies for two photon imaging

    Science.gov (United States)

    Mirkhanov, Shamil; Quarterman, Adrian H.; Smyth, Connor J. C. P.; Praveen, Bavishna B.; Appleton, Paul; Thomson, Calum; Swift, Samuel; Wilcox, Keith G.

    2017-02-01

    Multiphoton imaging (MPI) is an important fluorescence microscopy technique that allows deep tissue and in-vivo imaging with high selectivity. According to theory, two-photon signal is proportional to the product of the peak power and the average power, allowing optimization of key imaging parameters of the excitation laser, such as average power, repetition rate and pulse duration. Recent progress in compact ultrafast lasers including femtosecond fiber lasers and optically pumped semiconductor lasers makes direct control of these parameters possible. In order to investigate the optimum laser parameters for two photon imaging we experimentally study the effects of repetition rate between 2.85 and 90 MHz and pulse duration between 336 fs and 3.5 ps on two photon signal in SYTOX Green labeled mouse intestine sections at 1030 nm. We found that the optimum repetition rate for this sample is in the range 20 - 40 MHz, depending on average power, and that the pulse duration has no effect on the MPI signal provided that the average power can be adjusted to keep the product of average and peak power constant.

  13. Fast two-photon imaging of subcellular voltage dynamics in neuronal tissue with genetically encoded indicators.

    Science.gov (United States)

    Chamberland, Simon; Yang, Helen H; Pan, Michael M; Evans, Stephen W; Guan, Sihui; Chavarha, Mariya; Yang, Ying; Salesse, Charleen; Wu, Haodi; Wu, Joseph C; Clandinin, Thomas R; Toth, Katalin; Lin, Michael Z; St-Pierre, François

    2017-07-27

    Monitoring voltage dynamics in defined neurons deep in the brain is critical for unraveling the function of neuronal circuits but is challenging due to the limited performance of existing tools. In particular, while genetically encoded voltage indicators have shown promise for optical detection of voltage transients, many indicators exhibit low sensitivity when imaged under two-photon illumination. Previous studies thus fell short of visualizing voltage dynamics in individual neurons in single trials. Here, we report ASAP2s, a novel voltage indicator with improved sensitivity. By imaging ASAP2s using random-access multi-photon microscopy, we demonstrate robust single-trial detection of action potentials in organotypic slice cultures. We also show that ASAP2s enables two-photon imaging of graded potentials in organotypic slice cultures and in Drosophila . These results demonstrate that the combination of ASAP2s and fast two-photon imaging methods enables detection of neural electrical activity with subcellular spatial resolution and millisecond-timescale precision.

  14. Tracking T and B cells from two-photon microscopy imaging using constrained SMC clusters

    OpenAIRE

    Olivieri, David; Faro, José; Gómez-Conde, Iván; Tadokoro, Carlos E.

    2011-01-01

    This paper describes a novel software algorithm, called constrained Sequential Monte Carlo (SMC) clusters, for tracking a large collection of individual cells from intra-vital two-photon microscopy image sequences. We show how our method and software tool, implemented in python, is useful for quantifying the motility of T and B lymphocytes involved in an immune response vs lymphocytes under non immune conditions. We describe the theory behind our algorithm and briefly discuss the architecture...

  15. 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.

  16. Spatio-temporal imaging of surface plasmon polaritons in two photon photoemission microscopy

    Science.gov (United States)

    Meyer zu Heringdorf, Frank J.; Podbiel, Daniel; Raß, Nicolai; Makris, Andreas; Buckanie, Niemma M.; Kahl, Philip A.

    2016-09-01

    A two-photon photoemission microscopy experiment with femtosecond time-resolution for imaging of propagating surface plasmon polaritons is discussed. The experimental setup of an actively Pancharatnam's phase stabilized interferometer is described, and a temporal stability in time-resolved two-photon photoemission microscopy of less than 20 attoseconds is demonstrated. The time-resolved setup is applied to investigate the interaction of a surface plasmon polariton wave packet with a plasmonic beam-splitter. Pump-probe data recorded at times before and after the interaction of the surface plasmon polariton wave packet with the beam-splitter indicate transmission and reflection coefficients of T≍0.3 and R≍0.4, respectively.

  17. Deep two-photon brain imaging with a red-shifted fluorometric Ca2+ indicator.

    Science.gov (United States)

    Tischbirek, Carsten; Birkner, Antje; Jia, Hongbo; Sakmann, Bert; Konnerth, Arthur

    2015-09-08

    In vivo Ca2+ imaging of neuronal populations in deep cortical layers has remained a major challenge, as the recording depth of two-photon microscopy is limited because of the scattering and absorption of photons in brain tissue. A possible strategy to increase the imaging depth is the use of red-shifted fluorescent dyes, as scattering of photons is reduced at long wavelengths. Here, we tested the red-shifted fluorescent Ca2+ indicator Cal-590 for deep tissue experiments in the mouse cortex in vivo. In experiments involving bulk loading of neurons with the acetoxymethyl (AM) ester version of Cal-590, combined two-photon imaging and cell-attached recordings revealed that, despite the relatively low affinity of Cal-590 for Ca2+ (Kd=561 nM), single-action potential-evoked Ca2+ transients were discernable in most neurons with a good signal-to-noise ratio. Action potential-dependent Ca2+ transients were recorded in neurons of all six layers of the cortex at depths of up to -900 µm below the pial surface. We demonstrate that Cal-590 is also suited for multicolor functional imaging experiments in combination with other Ca2+ indicators. Ca2+ transients in the dendrites of an individual Oregon green 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-1 (OGB-1)-labeled neuron and the surrounding population of Cal-590-labeled cells were recorded simultaneously on two spectrally separated detection channels. We conclude that the red-shifted Ca2+ indicator Cal-590 is well suited for in vivo two-photon Ca2+ imaging experiments in all layers of mouse cortex. In combination with spectrally different Ca2+ indicators, such as OGB-1, Cal-590 can be readily used for simultaneous multicolor functional imaging experiments.

  18. Imaging immune response of skin mast cells in vivo with two-photon microscopy

    Science.gov (United States)

    Li, Chunqiang; Pastila, Riikka K.; Lin, Charles P.

    2012-02-01

    Intravital multiphoton microscopy has provided insightful information of the dynamic process of immune cells in vivo. However, the use of exogenous labeling agents limits its applications. There is no method to perform functional imaging of mast cells, a population of innate tissue-resident immune cells. Mast cells are widely recognized as the effector cells in allergy. Recently their roles as immunoregulatory cells in certain innate and adaptive immune responses are being actively investigated. Here we report in vivo mouse skin mast cells imaging with two-photon microscopy using endogenous tryptophan as the fluorophore. We studied the following processes. 1) Mast cells degranulation, the first step in the mast cell activation process in which the granules are released into peripheral tissue to trigger downstream reactions. 2) Mast cell reconstitution, a procedure commonly used to study mast cells functioning by comparing the data from wild type mice, mast cell-deficient mice, and mast-cell deficient mice reconstituted with bone marrow-derived mast cells (BMMCs). Imaging the BMMCs engraftment in tissue reveals the mast cells development and the efficiency of BMMCs reconstitution. We observed the reconstitution process for 6 weeks in the ear skin of mast cell-deficient Kit wsh/ w-sh mice by two-photon imaging. Our finding is the first instance of imaging mast cells in vivo with endogenous contrast.

  19. 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.

  20. An automated detection for axonal boutons in vivo two-photon imaging of mouse

    Science.gov (United States)

    Li, Weifu; Zhang, Dandan; Xie, Qiwei; Chen, Xi; Han, Hua

    2017-02-01

    Activity-dependent changes in the synaptic connections of the brain are tightly related to learning and memory. Previous studies have shown that essentially all new synaptic contacts were made by adding new partners to existing synaptic elements. To further explore synaptic dynamics in specific pathways, concurrent imaging of pre and postsynaptic structures in identified connections is required. Consequently, considerable attention has been paid for the automated detection of axonal boutons. Different from most previous methods proposed in vitro data, this paper considers a more practical case in vivo neuron images which can provide real time information and direct observation of the dynamics of a disease process in mouse. Additionally, we present an automated approach for detecting axonal boutons by starting with deconvolving the original images, then thresholding the enhanced images, and reserving the regions fulfilling a series of criteria. Experimental result in vivo two-photon imaging of mouse demonstrates the effectiveness of our proposed method.

  1. 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.

  2. 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.

  3. 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...

  4. 3-D IR imaging with uncooled GaN photodiodes using nondegenerate two-photon absorption

    CERN Document Server

    Pattanaik, Himansu S; Hagan, David J; Van Stryland, Eric W

    2015-01-01

    We utilize the recently demonstrated orders of magnitude enhancement of extremely nondegenerate two-photon absorption in direct-gap semiconductor photodiodes to perform scanned imaging of 3D structures using IR femtosecond illumination pulses (1.6 um and 4.93 um) gated on the GaN detector by sub-gap, femtosecond pulses. While transverse resolution is limited by the usual imaging criteria, the longitudinal or depth resolution can be less than a wavelength, dependent on the pulsewidths in this nonlinear interaction within the detector element. The imaging system can accommodate a wide range of wavelengths in the mid-IR and near-IR without the need to modify the detection and imaging systems.

  5. Multimodal imaging of lung tissue using optical coherence tomography and two photon microscopy

    Science.gov (United States)

    Gaertner, Maria; Cimalla, Peter; Geissler, Stefan; Meissner, Sven; Schnabel, Christian; Kuebler, Wolfgang M.; Koch, Edmund

    2012-02-01

    In the context of protective artificial ventilation strategies for patients with severe lung diseases, the contribution of ventilator settings to tissue changes on the alveolar level of the lung is still a question under debate. To understand the impact of respiratory settings as well as the dynamic process of respiration, high-resolution monitoring and visualization of the dynamics of lung alveoli are essential. An instrument allowing 3D imaging of lung tissue as well as imaging of functional constituents, such as elastin fibers, in in situ experimental conditions is presented in this study using a combination of Fourier domain optical coherence tomography (FD-OCT) and fiber-guided two photon microscopy. In a comparative study, fixed lung tissue, stained with sulforhodamine B for elastin fibers, was used to illustrate the ability of fiber-guided two photon excitation and single photon excitation for the visualization of elastin fibers within the tissue. Together with the fast 3D imaging capability of OCT, a new tool is given for the monitoring of alveolar lung dynamics in future in vivo experiments.

  6. Development of large field-of-view two photon microscopy for imaging mouse cortex (Conference Presentation)

    Science.gov (United States)

    Bumstead, Jonathan; Côté, Daniel C.; Culver, Joseph P.

    2017-02-01

    Spontaneous neuronal activity has been measured at cellular resolution in mice, zebrafish, and C. elegans using optical sectioning microscopy techniques, such as light sheet microscopy (LSM) and two photon microscopy (TPM). Recent improvements in these modalities and genetically encoded calcium indicators (GECI's) have enabled whole brain imaging of calcium dynamics in zebrafish and C. elegans. However, these whole brain microscopy studies have not been extended to mice due to the limited field of view (FOV) of TPM and the cumbersome geometry of LSM. Conventional TPM is restricted to diffraction limited imaging over this small FOV (around 500 x 500 microns) due to the use of high magnification objectives (e.g. 1.0 NA; 20X) and the aberrations introduced by relay optics used in scanning the beam across the sample. To overcome these limitations, we have redesigned the entire optical path of the two photon microscope (scanning optics and objective lens) to support a field of view of Ø7 mm with relatively high spatial resolution (engineering software Zemax, we designed our system with commercially available optics that minimize astigmatism, field curvature, chromatic focal shift, and vignetting. Performance of the system was also tested experimentally with fluorescent beads in agarose, fixed samples, and in vivo structural imaging. Our large-FOV TPM provides a modality capable of studying distributed brain networks in mice at cellular resolution.

  7. 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.

  8. Multiscale vision model for event detection and reconstruction in two-photon imaging data

    DEFF Research Database (Denmark)

    Brazhe, Alexey; Mathiesen, Claus; Lind, Barbara Lykke

    2014-01-01

    on a modified multiscale vision model, an object detection framework based on the thresholding of wavelet coefficients and hierarchical trees of significant coefficients followed by nonlinear iterative partial object reconstruction, for the analysis of two-photon calcium imaging data. The framework is discussed...... of the multiscale vision model is similar in the denoising, but provides a better segmenation of the image into meaningful objects, whereas other methods need to be combined with dedicated thresholding and segmentation utilities.......Reliable detection of calcium waves in multiphoton imaging data is challenging because of the low signal-to-noise ratio and because of the unpredictability of the time and location of these spontaneous events. This paper describes our approach to calcium wave detection and reconstruction based...

  9. Single pulse two photon fluorescence lifetime imaging (SP-FLIM) with MHz pixel rate.

    Science.gov (United States)

    Eibl, Matthias; Karpf, Sebastian; Weng, Daniel; Hakert, Hubertus; Pfeiffer, Tom; Kolb, Jan Philip; Huber, Robert

    2017-07-01

    Two-photon-excited fluorescence lifetime imaging microscopy (FLIM) is a chemically specific 3-D sensing modality providing valuable information about the microstructure, composition and function of a sample. However, a more widespread application of this technique is hindered by the need for a sophisticated ultra-short pulse laser source and by speed limitations of current FLIM detection systems. To overcome these limitations, we combined a robust sub-nanosecond fiber laser as the excitation source with high analog bandwidth detection. Due to the long pulse length in our configuration, more fluorescence photons are generated per pulse, which allows us to derive the lifetime with a single excitation pulse only. In this paper, we show high quality FLIM images acquired at a pixel rate of 1 MHz. This approach is a promising candidate for an easy-to-use and benchtop FLIM system to make this technique available to a wider research community.

  10. Imaging of surgical margin in pancreatic metastasis using two-photon excited fluorescence microscopy

    Science.gov (United States)

    Chen, Jing; Hong, Zhipeng; Chen, Hong; Chen, Youting; Xu, Yahao; Zhu, Xiaoqin; Zhuo, Shuangmu; Shi, Zheng; Chen, Jianxin

    2014-09-01

    Two-photon excited fluorescence (TPEF) microscopy, has become a powerful tool for imaging unstained tissue samples at subcellular level in biomedical research. The purpose of this study was to determine whether TPEF imaging of histological sections without H-E staining can be used to identify the boundary between normal pancreas and pancreatic metastasis from renal cell carcinoma (RCC). The typical features such as the significant increase of cancerous nests, the absence of pancreatic ductal, the appearance of cancer cells were observed to present the boundary between normal pancreas and pancreatic metastasis from RCC. These results correlated well with the corresponding histological outcomes. With the advent of clinically miniaturized TPEF microscopy and integrative endoscopy, TPEF microscopy has the potential application on surgical location of pancreatic metastasis from RCC in the near future.

  11. Two-photon fluorescence lifetime imaging of primed SNARE complexes in presynaptic terminals and β cells

    Science.gov (United States)

    Takahashi, Noriko; Sawada, Wakako; Noguchi, Jun; Watanabe, Satoshi; Ucar, Hasan; Hayashi-Takagi, Akiko; Yagishita, Sho; Ohno, Mitsuyo; Tokumaru, Hiroshi; Kasai, Haruo

    2015-10-01

    It remains unclear how readiness for Ca2+-dependent exocytosis depends on varying degrees of SNARE complex assembly. Here we directly investigate the SNARE assembly using two-photon fluorescence lifetime imaging (FLIM) of Förster resonance energy transfer (FRET) between three pairs of neuronal SNAREs in presynaptic boutons and pancreatic β cells in the islets of Langerhans. These FRET probes functionally rescue their endogenous counterparts, supporting ultrafast exocytosis. We show that trans-SNARE complexes accumulated in the active zone, and estimate the number of complexes associated with each docked vesicle. In contrast, SNAREs were unassembled in resting state, and assembled only shortly prior to insulin exocytosis, which proceeds slowly. We thus demonstrate that distinct states of fusion readiness are associated with SNARE complex formation. Our FRET/FLIM approaches enable optical imaging of fusion readiness in both live and chemically fixed tissues.

  12. Delivery of ultrashort spatially focused pulses through a multimode fiber for two photon endoscopic imaging

    Science.gov (United States)

    Morales-Delgado, Edgar E.; Papadopoulos, Ioannis N.; Farahi, Salma; Psaltis, Demetri; Moser, Christophe

    2015-06-01

    Due to their high number of supported modes, multimode optical fibers carry large amount of spatio-temporal information. However, propagation of a light pulse through a multimode optical fiber suffers from spatial distortions due to superposition of the various exited modes and from time broadening due to modal dispersion. Here, we present a method based on digital phase conjugation to selectively excite specific optical fiber modes in a multimode fiber that follow similar optical paths as they travel through the fiber. In this way, they can be made to interfere constructively at the fiber output to generate an ultrashort spatially focused pulse. The excitation of a limited number of modes limits modal dispersion, allowing the transmission of an ultrashort pulse. We also show that the short spatially focused pulse can be scanned digitally without movable elements. We experimentally demonstrate that the pulse at the output of the multimode fiber generate a two-photon signal. We show delivery of a 1550 nm pulse with 500 fs duration, spatially focused to a spot size of 7 micrometers, through a 30 cm long, 200 micrometers core multimode step-index fiber. We show how this technique is applied to endoscopic two-photon imaging.

  13. Paramagnetic, silicon quantum dots for magnetic resonance and two photon imaging of macrophages

    Science.gov (United States)

    Tu, Chuqiao; Ma, Xuchu; Pantazis, Periklis; Kauzlarich, Susan M.; Louie, Angelique Y.

    2010-01-01

    Quantum dots (QDs) are an attractive platform for building multimodality imaging probes, but the toxicity for typical cadmium QDs limits enthusiasm for their clinical use. Nontoxic, silicon QDs are more promising but tend to require short wavelength excitations which are subject to tissue scattering and autofluorescence artifacts. Herein, we report the synthesis of paramagnetic, manganese-doped, silicon QDs ((SiMn QDs) and demonstrate that they are detectable by both MRI and near infrared excited, two-photon imaging. The SiMn QDs are coated with dextran sulfate to target them to scavenger receptors on macrophages, a biomarker of vulnerable plaques. TEM images show that isolated QDs have an average core diameter of 4.3 ± 1.0 nm and the hydrodynamic diameters of coated nanoparticles range from 8.3 to 43 nm measured by Dynamic Light Scattering (DLS). The SiMn QDs have an r1 relaxivity of 25.50 ± 1.44 mM−1s−1 and an r2 relaxivity of 89.01 ± 3.26 mM−1s−1 (37 °C, 1.4 T). They emit strong fluorescence at 441 nm with a quantum yield of 8.1% in water. Cell studies show that the probes specifically accumulate in macrophages by a receptor-mediated process, are nontoxic to mammalian cells, and produce distinct contrast in both T1-weighted magnetic resonance and single- or two-photon excitation fluorescence images. These QDs have promising diagnostic potential as high macrophage density is associated with atherosclerotic plaques vulnerable to rupture. PMID:20092250

  14. Paramagnetic, silicon quantum dots for magnetic resonance and two-photon imaging of macrophages.

    Science.gov (United States)

    Tu, Chuqiao; Ma, Xuchu; Pantazis, Periklis; Kauzlarich, Susan M; Louie, Angelique Y

    2010-02-17

    Quantum dots (QDs) are an attractive platform for building multimodality imaging probes, but the toxicity for typical cadmium QDs limits enthusiasm for their clinical use. Nontoxic, silicon QDs are more promising but tend to require short-wavelength excitations which are subject to tissue scattering and autofluorescence artifacts. Herein, we report the synthesis of paramagnetic, manganese-doped, silicon QDs (Si(Mn) QDs) and demonstrate that they are detectable by both MRI and near-infrared excited, two-photon imaging. The Si(Mn) QDs are coated with dextran sulfate to target them to scavenger receptors on macrophages, a biomarker of vulnerable plaques. TEM images show that isolated QDs have an average core diameter of 4.3 +/- 1.0 nm and the hydrodynamic diameters of coated nanoparticles range from 8.3 to 43 nm measured by dynamic light scattering (DLS). The Si(Mn) QDs have an r(1) relaxivity of 25.50 +/- 1.44 mM(-1) s(-1) and an r(2) relaxivity of 89.01 +/- 3.26 mM(-1) s(-1) (37 degrees C, 1.4 T). They emit strong fluorescence at 441 nm with a quantum yield of 8.1% in water. Cell studies show that the probes specifically accumulate in macrophages by a receptor-mediated process, are nontoxic to mammalian cells, and produce distinct contrast in both T(1)-weighted magnetic resonance and single- or two-photon excitation fluorescence images. These QDs have promising diagnostic potential as high macrophage density is associated with atherosclerotic plaques vulnerable to rupture.

  15. Resolution of multiple GFP color variants and dyes using two-photon microscopy and imaging spectroscopy

    Science.gov (United States)

    Lansford, Rusty; Bearman, Gregory H.; Fraser, Scott E.

    2001-07-01

    The imaging of living cells and tissues using laser-scanning microscopy is offering dramatic insights into the spatial and temporal controls of biological processes. The availability of genetically encoded labels such as green fluorescent protein (GFP) offers unique opportunities by which to trace cell movements, cell signaling or gene expression dynamically in developing embryos. Two-photon laser scanning microscopy (TPLSM) is ideally suited to imaging cells in vivo due to its deeper tissue penetration and reduced phototoxicity; however, in TPLSM the excitation and emission spectra of GFP and its color variants [e.g., CyanFP (CFP); yellowFP (YFP)] are insufficiently distinct to be uniquely imaged by conventional means. To surmount such difficulties, we have combined the technologies of TPLSM and imaging spectroscopy to unambiguously identify CFP, GFP, YFP, and redFP (RFP) as well as conventional dyes, and have tested the approach in cell lines. In our approach, a liquid crystal tunable filter was used to collect the emission spectrum of each pixel within the TPLSM image. Based on the fluorescent emission spectra, supervised classification and linear unmixing analysis algorithms were used to identify the nature and relative amounts of the fluorescent proteins expressed in the cells. In a most extreme case, we have used the approach to separate GFP and fluorescein, separated by only 7 nm, and appear somewhat indistinguishable by conventional techniques. This approach offers the needed ability to concurrently image multiple colored, spectrally overlapping marker proteins within living cells.

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

    Science.gov (United States)

    Broess, Koen; Borst, Jan Willem; van Amerongen, Herbert

    2009-05-01

    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 Arabidopsis thaliana and Alocasia wentii under excitation-annihilation free conditions, both for the F (0)- and the F (m)-state. The corresponding average lifetimes are approximately 250 ps and approximately 1.5 ns, respectively, similar to those of isolated chloroplasts. These values appear to be the same for chloroplasts in the top, middle, and bottom layer of the leaves. With the spatial resolution of approximately 500 nm in the focal (xy) plane and 2 microm in the z direction, it appears to be impossible to fully resolve the grana stacks and stroma lamellae, but variations in the fluorescence lifetimes, and thus of the composition on a pixel-to-pixel base can be observed.

  17. Two-photon calcium imaging during fictive navigation in virtual environments

    Directory of Open Access Journals (Sweden)

    Misha Benjamin Ahrens

    2013-06-01

    Full Text Available A full understanding of nervous system function requires recording from large populations of neurons during naturalistic behaviors. Here we enable paralyzed larval zebrafish to fictively navigate two-dimensional virtual environments while we record optically from many neurons with two-photon imaging. Electrical recordings from motor nerves in the tail are decoded into intended forward swims and turns, which are used to update a virtual environment displayed underneath the fish. Several behavioral features - such as turning responses to whole-field motion and dark avoidance - are well-replicated in this virtual setting. We readily observed neuronal populations in the hindbrain with laterally selective responses that correlated with right or left optomotor behavior. We also observed neurons in the habenula, pallium, and midbrain with response properties specific to environmental features. Beyond single-cell correlations, the classification of network activity in such virtual settings promises to reveal principles of brainwide neural dynamics during behavior.

  18. Two-photon calcium imaging during fictive navigation in virtual environments.

    Science.gov (United States)

    Ahrens, Misha B; Huang, Kuo Hua; Narayan, Sujatha; Mensh, Brett D; Engert, Florian

    2013-01-01

    A full understanding of nervous system function requires recording from large populations of neurons during naturalistic behaviors. Here we enable paralyzed larval zebrafish to fictively navigate two-dimensional virtual environments while we record optically from many neurons with two-photon imaging. Electrical recordings from motor nerves in the tail are decoded into intended forward swims and turns, which are used to update a virtual environment displayed underneath the fish. Several behavioral features-such as turning responses to whole-field motion and dark avoidance-are well-replicated in this virtual setting. We readily observed neuronal populations in the hindbrain with laterally selective responses that correlated with right or left optomotor behavior. We also observed neurons in the habenula, pallium, and midbrain with response properties specific to environmental features. Beyond single-cell correlations, the classification of network activity in such virtual settings promises to reveal principles of brainwide neural dynamics during behavior.

  19. Tracking T and B cells from two-photon microscopy imaging using constrained SMC clusters.

    Science.gov (United States)

    Olivieri, D; Faro, J; Gomez-Conde, I; Tadokoro, C E

    2011-09-16

    This paper describes a novel software algorithm, called constrained Sequential Monte Carlo (SMC) clusters, for tracking a large collection of individual cells from intra-vital two-photon microscopy image sequences. We show how our method and software tool, implemented in python, is useful for quantifying the motility of T and B lymphocytes involved in an immune response vs lymphocytes under non immune conditions. We describe the theory behind our algorithm and briefly discuss the architecture of our software. Finally, we demonstrate both the functionality and utility of software by applying it to two practical examples from videos displaying lymphocyte motility in B cell zones (follicles) and T cell zones of lymph nodes. Copyright 2011 The Author(s). Published by Journal of Integrative Bioinformatics.

  20. Simultaneous two-photon imaging of cerebral oxygenation and capillary blood flow in atherosclerotic mice

    Science.gov (United States)

    Lu, Xuecong; Li, Baoqiang; Moeini, Mohammad; Lesage, Frédéric

    2017-02-01

    Gradual changes in brain microvasculature and cerebral capillary blood flow occurring with atherosclerosis may significantly contribute to cognition decline due to their role in brain tissue oxygenation. However, previous stud- ies of the relationship between cerebral capillary blood flow and brain tissue oxygenation are limited. This study aimed to investigate vascular and concomitant changes in brain tissue pO2 with atherosclerosis. Experiments in young healthy C57B1/6 mice (n=6 , WT), young atherosclerotic mice (n=6 , ATX Y) and old atherosclerotic mice (n=6 , ATX O) were performed imaging on the left sensory-motor cortex at resting state under urethane (1.5 g/kg) anesthesia using two-photon fluorescence microscopy. The results showed that pO2 around capillaries, correlated with red blood cell (RBC) flux, increased with atherosclerosis.

  1. A long Stokes shift red fluorescent Ca2+ indicator protein for two-photon and ratiometric imaging.

    Science.gov (United States)

    Wu, Jiahui; Abdelfattah, Ahmed S; Miraucourt, Loïs S; Kutsarova, Elena; Ruangkittisakul, Araya; Zhou, Hang; Ballanyi, Klaus; Wicks, Geoffrey; Drobizhev, Mikhail; Rebane, Aleksander; Ruthazer, Edward S; Campbell, Robert E

    2014-10-31

    The introduction of calcium ion (Ca(2+)) indicators based on red fluorescent proteins (RFPs) has created new opportunities for multicolour visualization of intracellular Ca(2+) dynamics. However, one drawback of these indicators is that they have optimal two-photon excitation outside the near-infrared window (650-1,000 nm) where tissue is most transparent to light. To address this shortcoming, we developed a long Stokes shift RFP-based Ca(2+) indicator, REX-GECO1, with optimal two-photon excitation at indicator in organotypic hippocampal slice cultures (one- and two-photon) and the visual system of albino tadpoles (two-photon). Furthermore, we demonstrate single excitation wavelength two-colour Ca(2+) and glutamate imaging in organotypic cultures.

  2. Automated 3-D Detection of Dendritic Spines from In Vivo Two-Photon Image Stacks.

    Science.gov (United States)

    Singh, P K; Hernandez-Herrera, P; Labate, D; Papadakis, M

    2017-10-01

    Despite the significant advances in the development of automated image analysis algorithms for the detection and extraction of neuronal structures, current software tools still have numerous limitations when it comes to the detection and analysis of dendritic spines. The problem is especially challenging in in vivo imaging, where the difficulty of extracting morphometric properties of spines is compounded by lower image resolution and contrast levels native to two-photon laser microscopy. To address this challenge, we introduce a new computational framework for the automated detection and quantitative analysis of dendritic spines in vivo multi-photon imaging. This framework includes: (i) a novel preprocessing algorithm enhancing spines in a way that they are included in the binarized volume produced during the segmentation of foreground from background; (ii) the mathematical foundation of this algorithm, and (iii) an algorithm for the detection of spine locations in reference to centerline trace and separating them from the branches to whom spines are attached to. This framework enables the computation of a wide range of geometric features such as spine length, spatial distribution and spine volume in a high-throughput fashion. We illustrate our approach for the automated extraction of dendritic spine features in time-series multi-photon images of layer 5 cortical excitatory neurons from the mouse visual cortex.

  3. Gold Nanorods-Based Theranostics for Simultaneous Fluorescence/Two-Photon Luminescence Imaging and Synergistic Phototherapies

    Directory of Open Access Journals (Sweden)

    Shan Fang

    2016-01-01

    Full Text Available Gold nanorods (GNRs have shown great potential applications in cancer theranostics due to the unique phenomenon of surface plasmon resonance, which leads to strong electric fields on the surface and consequently enhances the absorption and scattering in the near-infrared (NIR region. Indocyanine green (ICG, an amphipathic dye, is not only an excellent NIR imaging agent but also an ideal light absorber for laser-mediated photodynamic and photothermal therapy. In this study, in order to integrate the merits of GNRs and ICG in biomedical applications, we developed ICG conjugated silica-coated GNRs (GNR@SiO2-ICG for cancer imaging and phototherapy. The covalent coupling strategy reduces the probability of leakage/desorption during the delivery. The as-prepared GNR@SiO2-ICG could serve as efficient probes to simultaneously enhance fluorescence (FL imaging and two-photon luminescence (TPL imaging. In vitro experiments indicated that A375 cells could be killed through synergistic phototherapies effect of GNRs and ICG using single wavelength continuous-wave laser irradiation. Our results indicated that the synthesized GNR@SiO2-ICG are effective for simultaneously enhancing FL/TPL imaging and synergistic phototherapies.

  4. Nanosecond two-photon excitation fluorescence imaging with a multi color fiber MOPA laser

    Science.gov (United States)

    Karpf, Sebastian; Eibl, Matthias; Huber, Robert

    2015-07-01

    A system is presented that uses a fiber based Master Oscillator Power Amplifier (MOPA) with nanosecond-range pulses for two-photon excitation fluorescence (TPEF) imaging. The robust laser in the extended near infrared is based on an actively modulated electro-optical modulator (EOM), enabling free synchronization of the pulses to any other light source or detection unit. Pulses with a freely programmable duration between 0.4 and 10 ns are generated and then amplified to up to kilowatts of peak power with ytterbium doped fiber amplifiers (YDFA). Since we achieve peak power and duty cycles comparable to standard femto- and picosecond setups, the TPEF signal levels are similar, but realized with a robust and inexpensive fiber-based setup. The delivery fiber is further used as an optional, electronically controllable Raman shifter to effectively shift the 1064 nm light to 1122 nm and to 1186 nm. This allows imaging of a manifold of fluorophores, like e.g. TexasRed, mCherry, mRaspberry and many more. We show TPEF imaging of the autofluorescence of plant leaves of moss and algae, acquired in epi-direction. This modular laser unit can be integrated into existing systems as either a fiber-based, alignment free excitation laser or an extension for multi-modal imaging.

  5. Imaging living hair cells within the cochlear epithelium of mice using two-photon microscopy

    Science.gov (United States)

    Yuan, Tao; Gao, Simon S.; Saggau, Peter; Oghalai, John S.

    2009-02-01

    Mice are an excellent model for studying mammalian hearing and transgenic mouse models of human hearing loss are commonly available for research. However, the mouse cochlea is substantially smaller than other animal models routinely used to study cochlear physiology. This makes the study of their hair cells difficult. We developed a novel methodology to optically image calcium within living hair cells left undisturbed within the excised mouse cochlea. Fresh cochleae were harvested, left intact within their otic capsule bone, and glued upright in a recording chamber. The bone overlying the region of the cochlear epithelium to be studied was opened and Reissner's membrane was incised. A fluorescent indicator was applied to the preparation to image intracellular calcium. A custom-built upright two-photon microscope was used to image the preparation using three dimensional scanning. We were able to image about 1/3 of a cochlear turn simultaneously, in either the apical or basal regions. Within one hour of animal sacrifice, we found that outer hair cells demonstrated increased fluorescence compared with surrounding supporting cells. Thus, this methodology can be used to visualize hair cell calcium changes and mechanotransduction over a region of the epithelium. Because the epithelium is left within the cochlea, dissection trauma is minimized and artifactual changes in hair cell physiology are reduced.

  6. Label-free in vivo imaging of human leukocytes using two-photon excited endogenous fluorescence

    Science.gov (United States)

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

    2013-04-01

    We demonstrate that two-photon excited endogenous fluorescence enables label-free morphological and functional imaging of various human blood cells. Specifically, we achieved distinctive morphological contrast to visualize morphology of important leukocytes, such as polymorphonuclear structure of granulocyte and mononuclear feature of agranulocyte, through the employment of the reduced nicotinamide adenine dinucleotide (NADH) fluorescence signals. In addition, NADH fluorescence images clearly reveal the morphological transformation process of neutrophils during disease-causing bacterial infection. Our findings also show that time-resolved NADH fluorescence can be potentially used for functional imaging of the phagocytosis of pathogens by leukocytes (neutrophils) in vivo. In particular, we found that free-to-bound NADH ratios measured in infected neutrophils increased significantly, which is consistent with a previous study that the energy consumed in the phagocytosis of neutrophils is mainly generated through the glycolysis pathway that leads to the accumulation of free NADH. Future work will focus on further developing and applying label-free imaging technology to investigate leukocyte-related diseases and disorders.

  7. A two-photon fluorescent probe with a large turn-on signal for imaging hydrogen sulfide in living tissues

    Energy Technology Data Exchange (ETDEWEB)

    Zheng, Kaibo [State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, Hunan 410082 (China); Lin, Weiying, E-mail: weiyinglin2013@163.com [Institute of Fluorescent Probes for Biological Imaging, University of Jinan, Jinan, Shandong 250022 (China); State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, Hunan 410082 (China); Tan, Li; Cheng, Dan [State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, Hunan 410082 (China)

    2015-01-01

    Highlights: • A two-photon fluorescent probe for sensing H{sub 2}S was developed. • The probe shows a large turn on signal (120-fold enhancement). • The probe is suitable for fluorescence imaging of H{sub 2}S in living cells and tissues. • The probe was capable of detecting H{sub 2}S up to 170 μm depth in live tissues. - Abstract: A two-photon fluorescence turn-on H{sub 2}S probe GCTPOC–H{sub 2}S based on a two-photon platform with a large cross-section, GCTPOC, and a sensitive H{sub 2}S recognition site, dinitrophenyl ether was constructed. The probe GCTPOC–H{sub 2}S exhibits desirable properties such as high sensitivity, high selectivity, functioning well at physiological pH and low cytotoxicity. In particular, the probe shows a 120-fold enhancement in the presence of Na{sub 2}S (500 μM), which is larger than the reported two-photon fluorescent H{sub 2}S probes. The large fluorescence enhancement of the two-photon probe GCTPOC–H{sub 2}S renders it attractive for imaging H{sub 2}S in living tissues with deep tissue penetration. Significantly, we have demonstrated that the probe GCTPOC–H{sub 2}S is suitable for fluorescence imaging of H{sub 2}S in living tissues with deep penetration by using two-photon microscopy. The further application of the two-photon probe for the investigation of biological functions and pathological roles of H{sub 2}S in living systems is under progress.

  8. Measurement of drug-target engagement in live cells by two-photon fluorescence anisotropy imaging.

    Science.gov (United States)

    Vinegoni, Claudio; Fumene Feruglio, Paolo; Brand, Christian; Lee, Sungon; Nibbs, Antoinette E; Stapleton, Shawn; Shah, Sunil; Gryczynski, Ignacy; Reiner, Thomas; Mazitschek, Ralph; Weissleder, Ralph

    2017-07-01

    The ability to directly image and quantify drug-target engagement and drug distribution with subcellular resolution in live cells and whole organisms is a prerequisite to establishing accurate models of the kinetics and dynamics of drug action. Such methods would thus have far-reaching applications in drug development and molecular pharmacology. We recently presented one such technique based on fluorescence anisotropy, a spectroscopic method based on polarization light analysis and capable of measuring the binding interaction between molecules. Our technique allows the direct characterization of target engagement of fluorescently labeled drugs, using fluorophores with a fluorescence lifetime larger than the rotational correlation of the bound complex. Here we describe an optimized protocol for simultaneous dual-channel two-photon fluorescence anisotropy microscopy acquisition to perform drug-target measurements. We also provide the necessary software to implement stream processing to visualize images and to calculate quantitative parameters. The assembly and characterization part of the protocol can be implemented in 1 d. Sample preparation, characterization and imaging of drug binding can be completed in 2 d. Although currently adapted to an Olympus FV1000MPE microscope, the protocol can be extended to other commercial or custom-built microscopes.

  9. 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.

  10. 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.

  11. A flow bioreactor system compatible with real-time two-photon fluorescence lifetime imaging microscopy.

    Science.gov (United States)

    Shen, Nian; Riedl, Julia A; Carvajal-Berrio, Daniel A; Davis, Zackary; Monaghan, Michael G; Layland, Shannon Lee; Hinderer, Svenja; Schenke-Layland, Katja

    2017-11-17

    Bioreactors are essential cell and tissue culture tools that allow the introduction of biophysical signals into in vitro cultures. One major limitation is the need to interrupt experiments and sacrifice samples at certain time points for analyses. To address this issue, we designed a bioreactor that combines high-resolution contact-free imaging and continuous flow in a closed system that is compatible with various types of microscopes. The high-throughput fluid flow bioreactor was combined with two-photon fluorescence lifetime imaging microscopy (2P-FLIM) and validated. The hydrodynamics of the bioreactor chamber were characterized using COMSOL. The simulation of shear stress indicated that the bioreactor system provides homogeneous and reproducible flow conditions. The designed bioreactor was used to investigate the effects of low shear stress on human umbilical vein endothelial cells (HUVECs). In a scratch assay, we observed decreased migration of HUVECs under shear stress conditions. Furthermore, metabolic activity shifts from glycolysis to oxidative phosphorylation-dependent mechanisms in HUVECs cultured under low shear stress conditions were detected using 2P-FLIM. Future applications for this bioreactor range from observing cell fate development in real-time to monitoring the environmental effects on cells or metabolic changes due to drug applications. Creative Commons Attribution license.

  12. Visualizing heterogeneity of photosynthetic properties of plant leaves with two-photon fluorescence lifetime imaging microscopy.

    Science.gov (United States)

    Iermak, Ievgeniia; Vink, Jochem; Bader, Arjen N; Wientjes, Emilie; van Amerongen, Herbert

    2016-09-01

    Two-photon fluorescence lifetime imaging microscopy (FLIM) was used to analyse the distribution and properties of Photosystem I (PSI) and Photosystem II (PSII) in palisade and spongy chloroplasts of leaves from the C3 plant Arabidopsis thaliana and the C4 plant Miscanthus x giganteus. This was achieved by separating the time-resolved fluorescence of PSI and PSII in the leaf. It is found that the PSII antenna size is larger on the abaxial side of A. thaliana leaves, presumably because chloroplasts in the spongy mesophyll are "shaded" by the palisade cells. The number of chlorophylls in PSI on the adaxial side of the A. thaliana leaf is slightly higher. The C4 plant M. x giganteus contains both mesophyll and bundle sheath cells, which have a different PSI/PSII ratio. It is shown that the time-resolved fluorescence of bundle sheath and mesophyll cells can be analysed separately. The relative number of chlorophylls, which belong to PSI (as compared to PSII) in the bundle sheath cells is at least 2.5 times higher than in mesophyll cells. FLIM is thus demonstrated to be a useful technique to study the PSI/PSII ratio and PSII antenna size in well-defined regions of plant leaves without having to isolate pigment-protein complexes. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Simultaneous imaging of multiple focal planes in scanning two-photon absorption microscope by photon counting

    Science.gov (United States)

    Carriles, Ramón; Hoover, Erich E.; Amir, Wafa; Squier, Jeffery A.

    2007-09-01

    We demonstrate a two-photon absorption scanning microscope capable of imaging two focal planes simultaneously. The 23MHz fundamental laser is split in two, one part delayed in time while the other is focused with a deformable mirror to change its divergence. Both parts are then recombined to form a 46MHz pulse train consisting of two interlaced trains with different divergences that after the objective are focused at different sample depths. At the detection path, photon counting techniques allow photons coming from each depth to be separated based on their relative timing with respect to the 46MHz train. The separation is accomplished using a field-programmable gate array that has been programmed to switch back and forth between two counters at a rate provided by a master clock generated by the 46MHz pulse train. The computer that controls the scanners reads and resets the counters before moving to a new pixel. The scheme is demonstrated for two depths but can be extended to a larger number, the ultimate limit being the fluorescence lifetime. This technique could also be implemented for second or third harmonic generation microscopy, in this case the ultimate achievable number of focal planes would be determined by the electronics speed.

  14. 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.

  15. Two-photon imaging of cerebral hemodynamics and neural activity in awake and anesthetized marmosets.

    Science.gov (United States)

    Santisakultarm, Thom P; Kersbergen, Calvin J; Bandy, Daryl K; Ide, David C; Choi, Sang-Ho; Silva, Afonso C

    2016-09-15

    Marmosets are a powerful, emerging model for human behavior and neurological disorders. However, longitudinal imaging modalities that visualize both cellular structure and function within the cortex are not available in this animal model. Hence, we implemented an approach to quantify vascular topology, hemodynamics, and neural activity in awake marmosets using two-photon microscopy (2PM). Marmosets were acclimated to a custom stereotaxic system. AAV1-GCaMP5G was injected into somatosensory cortex to optically indicate neural activity, and a cranial chamber was implanted. Longitudinal 2PM revealed vasculature and neurons 500μm below the cortical surface. Vascular response and neural activity during sensory stimulation were preserved over 5 and 3 months, respectively, before optical quality deteriorated. Vascular remodeling including increased tortuosity and branching was quantified. However, capillary connectivity from arterioles to venules remained unchanged. Further, behavioral assessment before and after surgery demonstrated no impact on cognitive and motor function. Immunohistochemistry confirmed minimal astrocyte activation with no focal damage. Over 6 months, total cortical depth visualized decreased. When under anesthesia, the most prominent isoflurane-induced vasodilation occurred in capillaries and smaller arterioles. These results demonstrate the capability to repeatedly observe cortical physiology in awake marmosets over months. This work provides a novel and insightful technique to investigate critical mechanisms in neurological disorders in awake marmosets without introducing confounds from anesthesia. Published by Elsevier B.V.

  16. Review: two-photon scanning systems for clinical high resolution in vivo tissue imaging

    Science.gov (United States)

    König, K.; Müller, J.; Höfer, M.; Müller, C.; Weinigel, M.; Bückle, R.; Elsner, P.; Kaatz, M.; Messerschmidt, B.

    2008-02-01

    The femtosecond laser multiphoton tomograph DermaInspect as well as high NA two-photon GRIN microendoscopes for in vivo tomography of human skin have been used to detect malignant melanoma as well as to study the diffusion and intradermal accumulation of topically applied cosmetical and pharmaceutical components. So far, more than 500 patients and volunteers in Europe, Australia, and Asia have been investigated with this unique tomograph. Near infrared 80 MHz picojoule femtosecond laser pulses were employed to excite endogenous fluorophores such as NAD(P)H, flavoproteins, melanin, and elastin as well as fluorescent components of a variety of ointments via a twophoton excitation process. In addition, collagen has been imaged by second harmonic generation. Using a two-PMT detection system, the ratio of elastin to collagen was determined during optical sectioning. A high submicron spatial resolution and 50 picosecond temporal resolution was achieved using galvoscan mirrors and piezodriven focusing optics as well as a time-correlated single photon counting module with a fast microchannel plate detector and fast photomultipliers. Individual intratissue cells, mitochondria, melanosomes, and the morphology of the nuclei as well as extracellular matrix elements could be clearly visualized due to molecular imaging and the calculation of fluorescence lifetime images. Nanoparticles and intratissue drugs have been detected non-invasively, in situ and over a period of up to 3 months. In addition, hydration effects and UV effects were studied by monitoring modifications of cellular morphology and autofluorescence. The system was used to observe the diffusion through the stratum corneum and the accumulation and release of functionalized nanoparticles along hair shafts and epidermal ridges. The DermaInspect been also employed to gain information on skin age and wound healing in patients with ulcers. Novel developments include a galvo/piezo-scan driven flexible articulated arm as

  17. Automated biochemical, morphological, and organizational assessment of precancerous changes from endogenous two-photon fluorescence images.

    Directory of Open Access Journals (Sweden)

    Jonathan M Levitt

    Full Text Available Multi-photon fluorescence microscopy techniques allow for non-invasive interrogation of live samples in their native environment. These methods are particularly appealing for identifying pre-cancers because they are sensitive to the early changes that occur on the microscopic scale and can provide additional information not available using conventional screening techniques.In this study, we developed novel automated approaches, which can be employed for the real-time analysis of two-photon fluorescence images, to non-invasively discriminate between normal and pre-cancerous/HPV-immortalized engineered tissues by concurrently assessing metabolic activity, morphology, organization, and keratin localization. Specifically, we found that the metabolic activity was significantly enhanced and more uniform throughout the depths of the HPV-immortalized epithelia, based on our extraction of the NADH and FAD fluorescence contributions. Furthermore, we were able to separate the keratin contribution from metabolic enzymes to improve the redox estimates and to use the keratin localization as a means to discriminate between tissue types. To assess morphology and organization, Fourier-based, power spectral density (PSD approaches were employed. The nuclear size distribution throughout the epithelial depths was quantified by evaluating the variance of the corresponding spatial frequencies, which was found to be greater in the normal tissue compared to the HPV-immortalized tissues. The PSD was also used to calculate the Hurst parameter to identify the level of organization in the tissues, assuming a fractal model for the fluorescence intensity fluctuations within a field. We found the range of organization was greater in the normal tissue and closely related to the level of differentiation.A wealth of complementary morphological, biochemical and organizational tissue parameters can be extracted from high resolution images that are acquired based entirely on

  18. Automated biochemical, morphological, and organizational assessment of precancerous changes from endogenous two-photon fluorescence images.

    Science.gov (United States)

    Levitt, Jonathan M; McLaughlin-Drubin, Margaret E; Münger, Karl; Georgakoudi, Irene

    2011-01-01

    Multi-photon fluorescence microscopy techniques allow for non-invasive interrogation of live samples in their native environment. These methods are particularly appealing for identifying pre-cancers because they are sensitive to the early changes that occur on the microscopic scale and can provide additional information not available using conventional screening techniques. In this study, we developed novel automated approaches, which can be employed for the real-time analysis of two-photon fluorescence images, to non-invasively discriminate between normal and pre-cancerous/HPV-immortalized engineered tissues by concurrently assessing metabolic activity, morphology, organization, and keratin localization. Specifically, we found that the metabolic activity was significantly enhanced and more uniform throughout the depths of the HPV-immortalized epithelia, based on our extraction of the NADH and FAD fluorescence contributions. Furthermore, we were able to separate the keratin contribution from metabolic enzymes to improve the redox estimates and to use the keratin localization as a means to discriminate between tissue types. To assess morphology and organization, Fourier-based, power spectral density (PSD) approaches were employed. The nuclear size distribution throughout the epithelial depths was quantified by evaluating the variance of the corresponding spatial frequencies, which was found to be greater in the normal tissue compared to the HPV-immortalized tissues. The PSD was also used to calculate the Hurst parameter to identify the level of organization in the tissues, assuming a fractal model for the fluorescence intensity fluctuations within a field. We found the range of organization was greater in the normal tissue and closely related to the level of differentiation. A wealth of complementary morphological, biochemical and organizational tissue parameters can be extracted from high resolution images that are acquired based entirely on endogenous sources of

  19. Fluorenyl benzothiadiazole and benzoselenadiazole near-IR fluorescent probes for two-photon fluorescence imaging (Conference Presentation)

    Science.gov (United States)

    Belfield, Kevin D.; Yao, Sheng; Kim, Bosung; Yue, Xiling

    2016-03-01

    Imaging biological samples with two-photon fluorescence (2PF) microscopy has the unique advantage of resulting high contrast 3D resolution subcellular image that can reach up to several millimeters depth. 2PF probes that absorb and emit at near IR region need to be developed. Two-photon excitation (2PE) wavelengths are less concerned as 2PE uses wavelengths doubles the absorption wavelength of the probe, which means 2PE wavelengths for probes even with absorption at visible wavelength will fall into NIR region. Therefore, probes that fluoresce at near IR region with high quantum yields are needed. A series of dyes based on 5-thienyl-2, 1, 3-benzothiadiazole and 5-thienyl-2, 1, 3-benzoselenadiazole core were synthesized as near infrared two-photon fluorophores. Fluorescence maxima wavelengths as long as 714 nm and fluorescence quantum yields as high as 0.67 were achieved. The fluorescence quantum yields of the dyes were nearly constant, regardless of solvents polarity. These diazoles exhibited large Stokes shift (fluorescence figure of merit (FM , 1.04×10-2 GM). Cells incubated on a 3D scaffold with one of the new probes (encapsulated in Pluronic micelles) exhibited bright fluorescence, enabling 3D two-photon fluorescence imaging to a depth of 100 µm.

  20. Confocal reflectance and two-photon microscopy studies of a songbird skull for preparation of transcranial imaging

    Science.gov (United States)

    Abi-Haidar, Darine; Oliver, Thomas

    2009-05-01

    We present experiments and analyses of confocal reflectance and two-photon microscopy studies of zebra finch skull samples. The thin and hollow structure of these birds' skulls is quite translucent, which can allow in vivo transcranial two-photon imaging for brain activation monitoring. However, the skull structure is also quite complex, with high refractive index changes on a macroscopic scale. These studies aim at exploring the geometrical and scattering properties of these skull samples with the use of several confocal microscopy contrasts. Moreover, the study of the axial reflectance exponential decay is used to estimate the scattering coefficients of the bone. Finally, two-photon imaging experiments of a fluorescent object located beneath the skull are carried out. It reveals that two-photon fluorescence can be collected through the skull with a strong signal. It also reveals that the spatial resolution loss is quite high and cannot be fully explained by the bulk scattering properties of the bone, but also by the presence of the high refractive index inhomogeneity of this pneumatic skull structure. Even if the optical properties of the skull are different during in vivo experiments, these preliminary studies are aimed at preparing and optimizing transcranial brain activation monitoring experiments on songbirds.

  1. Efficient two-photon fluorescent probe with red emission for imaging of thiophenols in living cells and tissues.

    Science.gov (United States)

    Liu, Hong-Wen; Zhang, Xiao-Bing; Zhang, Jing; Wang, Qian-Qian; Hu, Xiao-Xiao; Wang, Peng; Tan, Weihong

    2015-09-01

    Thiophenols, a class of highly toxic and pollutant compounds, are widely used in industrial production. Some aliphatic thiols play important roles in living organisms. Therefore, the development of efficient methods to discriminate thiophenols from aliphatic thiols is of great importance. Although several one-photon fluorescent probes have been reported for thiophenols, two-photon fluorescent probes are more favorable for biological imaging due to its low background fluorescence, deep penetration depth, and so on. In this work, a two-photon fluorescent probe for thiophenols, termed NpRb1, has been developed for the first time by employing 2,4-dinitrobenzene-sulfonate (DNBS) as a recognition unit (also a fluorescence quencher) and a naphthalene-BODIPY-based through-bond energy transfer (TBET) cassette as a fluorescent reporter. The TBET system consists of a D-π-A structured two-photon naphthalene fluorophore and a red-emitting BODIPY. It displayed highly energy transfer efficiency (93.5%), large pseudo-Stokes shifts upon one-photon excitation, and red fluorescence emission (λem = 586 nm), which is highly desirable for bioimaging applications. The probe exhibited a 163-fold thiophenol-triggered two-photon excited fluorescence enhancement at 586 nm. It showed a high selectivity and excellent sensitivity to thiophenols, with a detection limit of 4.9 nM. Moreover, it was successfully applied for practical detection of thiophenol in water samples with a good recovery, two-photon imaging of thiophenol in living cells, and tissues with tissue-imaging depths of 90-220 μm, demonstrating its practical application in environmental samples and biological systems.

  2. 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.

  3. Two-photon imaging of field enhancement by groups of gold nanostrip antennas

    DEFF Research Database (Denmark)

    Novikov, Sergey M.; Beermann, Jonas; Sondergaard, Thomas

    2009-01-01

    Resonant field enhancement by groups of 16 nm thin gold nanostrip antennas consisting of four strips (widths of 70, 100, and 130 nm) and fixed gap (50, 100, 150, or 200 nm) between them and positioned on a quartz substrate is investigated by reflection spectroscopy and two-photon photoluminescence...... scanning optical microscopy. We obtain good agreement with theoretical calculated reflection spectra and the investigated two-photon intensity dependence on the nanoantenna configurations and parameters of the illumination showed field enhancements increasing with increasing gaps in each group for the p......-polarized (resonant) excitation, whereas these enhancement effects were not observed for the s-polarization (nonresonant) excitation. (C) 2009 Optical Society of America...

  4. Imaging of Fluoride Ion in Living Cells and Tissues with a Two-Photon Ratiometric Fluorescence Probe

    Directory of Open Access Journals (Sweden)

    Xinyue Zhu

    2015-01-01

    Full Text Available A reaction-based two-photon (TP ratiometric fluorescence probe Z2 has been developed and successfully applied to detect and image fluoride ion in living cells and tissues. The Z2 probe was designed designed to utilize an ICT mechanism between n-butylnaphthalimide as a fluorophore and tert-butyldiphenylsilane (TBDPS as a response group. Upon addition of fluoride ion, the Si-O bond in the Z2 would be cleaved, and then a stronger electron-donating group was released. The fluorescent changes at 450 and 540 nm, respectively, made it possible to achieve ratiometric fluorescence detection. The results indicated that the Z2 could ratiometrically detect and image fluoride ion in living cells and tissues in a depth of 250 μm by two-photon microscopy (TPM.

  5. A two-photon fluorescent probe for ratiometric imaging of endogenous hypochlorous acid in live cells and tissues.

    Science.gov (United States)

    Jun, Yong Woong; Sarkar, Sourav; Singha, Subhankar; Reo, Ye Jin; Kim, Hye Rim; Kim, Jong-Jin; Chang, Young-Tae; Ahn, Kyo Han

    2017-09-28

    A fluorescent probe that enables ratiometric imaging of endogenous hypochlorous acid (HOCl) in cells and tissues by two-photon microscopy is developed based on a red-emitting acetyl-benzocoumarin (AcBC) dye. An oxathiolane group in the probe reacts with HOCl to generate the AcBC dye, which involves a ratiometric fluorescence change only toward HOCl along with high sensitivity.

  6. Cross-sectional imaging of pendeo-epitaxial GaN using continuous-wave two-photon microphotoluminescence

    Science.gov (United States)

    Schuck, P. J.; Grober, R. D.; Roskowski, A. M.; Einfeldt, S.; Davis, R. F.

    2002-09-01

    A technique utilizing continuous-wave two-photon absorption has been developed for optically sectioning and imaging deep into GaN structures. Imaging at depths greater than 20 mum below the surface of a coalesced pendeo-epitaxial GaN sample is demonstrated. Free and donor-bound excitonic emission in this sample appears to originate at the surface, acceptor-bound exciton transitions are strongest in the top bulk portion of the sample, and subgap luminescence is most intense deep in the sample. The depth resolution of the imaging system is measured to be 1.75 mum near the GaN surface.

  7. Noninvasive corneal stromal collagen imaging using two-photon-generated second-harmonic signals.

    Science.gov (United States)

    Morishige, Naoyuki; Petroll, W Matthew; Nishida, Teruo; Kenney, M Cristina; Jester, James V

    2006-11-01

    To investigate the feasibility of using femtosecond-pulse lasers to produce second-harmonic generated (SHG) signals to noninvasively assess corneal stromal collagen organization. The Eye Institute, University of California, Irvine, California, USA. Mouse, rabbit, and human corneas were examined by two-photon confocal microscopy using a variable-wavelength femtosecond lasers to produce SHG signals. Two types were detected: forward scattered and backward scattered. Wavelength dependence of the SHG signal was confirmed by spectral separation using the 510 Meta (Zeiss). To verify the spatial relation between SHG signals and corneal cells, staining of cytoskeletons and nuclei was performed. Second-harmonic-generated signal intensity was strongest with an excitation wavelength of 800 nm for all 3 species. Second-harmonic-generated forward signals showed a distinct fibrillar pattern organized into bands suggesting lamellae, while backscattered SHG signals appeared more diffuse and indistinct. Reconstruction of SHG signals showed two patterns of lamellar organization: highly interwoven in the anterior stroma and orthogonally arranged in the posterior stroma. Unique to the human cornea was the presence of transverse, sutural lamellae that inserted into Bowman's layer, suggesting an anchoring function. Using two-photon confocal microscopy to generate SHG signals from the corneal collagen provides a powerful new approach to noninvasively study corneal structure. Human corneas had a unique organizational pattern with sutural lamellae to provide important biomechanical support that was not present in mouse or rabbit corneas.

  8. Fourth-order oriented partial-differential equations for noise removal of two-photon fluorescence images.

    Science.gov (United States)

    Wang, Yu; Ji, Xiangyang; Dai, Qionghai

    2010-09-01

    We derive the fourth-order oriented partial-differential equations (PDEs) for noise removal of two-photon fluorescence images. We consider it from two aspects: one is based on a variational method; the other is based on controlling the diffusion direction. Our filtering model makes the diffusion along only the special orientation--decided via all the information in the established filtering window, so the edges are protected during filtering. Compared with related PDEs models, our model shows superior performance in terms of both objective criteria and subjective human vision via processing simulated and experimental noisy images.

  9. Deep-red polymer dots with bright two-photon fluorescence and high biocompatibility for in vivo mouse brain imaging

    Science.gov (United States)

    Alifu, Nuernisha; Sun, Zezhou; Zebibula, Abudureheman; Zhu, Zhenggang; Zhao, Xinyuan; Wu, Changfeng; Wang, Yalun; Qian, Jun

    2017-09-01

    With high contrast and deep penetration, two-photon fluorescence (2PF) imaging has become one of the most promising in vivo fluorescence imaging techniques. To obtain good imaging contrast, fluorescent nanoprobes with good 2PF properties are highly needed. In this work, bright 2PF polymer dots (P dots) were applied for in vivo mouse brain imaging. Deep-red emissive P dots with PFBT as the donor and PFDBT5 as the acceptor were synthesized and used as a contrast agent. P dots were further encapsulated by poly(styrene-co-maleic anhydride) (PSMA) and grafted with poly(ethylene glycol) (PEG). The P dots-PEG exhibit large two-photon absorption (2PA) cross-sections (δ≥8500 g), good water dispersibility, and high biocompatibility. P dots-PEG was further utilized first time for in vivo vascular imaging of mouse ear and brain, under 690-900 nm femtosecond (fs) laser excitation. Due to the large 2PA cross-section and deep-red emission, a large imaging depth ( 720 μm) was achieved.

  10. Two-photon instant structured illumination microscopy improves the depth penetration of super-resolution imaging in thick scattering samples.

    Science.gov (United States)

    Winter, Peter W; York, Andrew G; Nogare, Damian Dalle; Ingaramo, Maria; Christensen, Ryan; Chitnis, Ajay; Patterson, George H; Shroff, Hari

    2014-09-20

    Fluorescence imaging methods that achieve spatial resolution beyond the diffraction limit (super-resolution) are of great interest in biology. We describe a super-resolution method that combines two-photon excitation with structured illumination microscopy (SIM), enabling three-dimensional interrogation of live organisms with ~150 nm lateral and ~400 nm axial resolution, at frame rates of ~1 Hz. By performing optical rather than digital processing operations to improve resolution, our microscope permits super-resolution imaging with no additional cost in acquisition time or phototoxicity relative to the point-scanning two-photon microscope upon which it is based. Our method provides better depth penetration and inherent optical sectioning than all previously reported super-resolution SIM implementations, enabling super-resolution imaging at depths exceeding 100 μm from the coverslip surface. The capability of our system for interrogating thick live specimens at high resolution is demonstrated by imaging whole nematode embryos and larvae, and tissues and organs inside zebrafish embryos.

  11. 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

  12. FocusStack and StimServer: a new open source MATLAB toolchain for visual stimulation and analysis of two-photon calcium neuronal imaging data.

    Science.gov (United States)

    Muir, Dylan R; Kampa, Björn M

    2014-01-01

    Two-photon calcium imaging of neuronal responses is an increasingly accessible technology for probing population responses in cortex at single cell resolution, and with reasonable and improving temporal resolution. However, analysis of two-photon data is usually performed using ad-hoc solutions. To date, no publicly available software exists for straightforward analysis of stimulus-triggered two-photon imaging experiments. In addition, the increasing data rates of two-photon acquisition systems imply increasing cost of computing hardware required for in-memory analysis. Here we present a Matlab toolbox, FocusStack, for simple and efficient analysis of two-photon calcium imaging stacks on consumer-level hardware, with minimal memory footprint. We also present a Matlab toolbox, StimServer, for generation and sequencing of visual stimuli, designed to be triggered over a network link from a two-photon acquisition system. FocusStack is compatible out of the box with several existing two-photon acquisition systems, and is simple to adapt to arbitrary binary file formats. Analysis tools such as stack alignment for movement correction, automated cell detection and peri-stimulus time histograms are already provided, and further tools can be easily incorporated. Both packages are available as publicly-accessible source-code repositories.

  13. Multimodal wide-field two-photon excitation imaging: characterization of the technique for in vivo applications.

    Science.gov (United States)

    Hwang, Jae Youn; Wachsmann-Hogiu, Sebastian; Ramanujan, V Krishnan; Nowatzyk, Andreas G; Koronyo, Yosef; Medina-Kauwe, Lali K; Gross, Zeev; Gray, Harry B; Farkas, Daniel L

    2011-01-13

    We report fast, non-scanning, wide-field two-photon fluorescence excitation with spectral and lifetime detection for in vivo biomedical applications. We determined the optical characteristics of the technique, developed a Gaussian flat-field correction method to reduce artifacts resulting from non-uniform excitation such that contrast is enhanced, and showed that it can be used for ex vivo and in vivo cellular-level imaging. Two applications were demonstrated: (i) ex vivo measurements of beta-amyloid plaques in retinas of transgenic mice, and (ii) in vivo imaging of sulfonated gallium(III) corroles injected into tumors. We demonstrate that wide-field two photon fluorescence excitation with flat-field correction provides more penetration depth as well as better contrast and axial resolution than the corresponding one-photon wide field excitation for the same dye. Importantly, when this technique is used together with spectral and fluorescence lifetime detection modules, it offers improved discrimination between fluorescence from molecules of interest and autofluorescence, with higher sensitivity and specificity for in vivo applications.

  14. 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.

  15. Lighting the Way to See Inside Two-Photon Absorption Materials: Structure–Property Relationship and Biological Imaging

    Science.gov (United States)

    Zhang, Qiong; Tian, Xiaohe; Zhou, Hongping; Wu, Jieying; Tian, Yupeng

    2017-01-01

    ions, including transition metals and lanthanides, can serve as an important part of the structure to control the intramolecular charge-transfer process that drives the 2PA process. As templates, transition metal ions can assemble simple to more sophisticated ligands in a variety of multipolar arrangements resulting in interesting and tailorable electronic and optical properties, depending on the nature of the metal center and the energetics of the metal-ligand interactions, such as intraligand charge-transfer (ILCT) and metal-ligand charge-transfer (MLCT) processes. Lanthanide complexes are attractive for a number of reasons: (i) their visible emissions are quite long-lived; (ii) their absorption and emission can be tuned with the aid of appropriate photoactive ligands; (iii) the accessible energy-transfer path between the photo-active ligands and the lanthanide ion can facilitate efficient lanthanide-based 2PA properties. Thus, the above materials with excellent 2PA properties should be applied in two-photon applications, especially two-photon fluorescence microscopy (TPFM) and related emission-based applications. Furthermore, the progress of research into the use of those new 2PA materials with moderate 2PA cross section in the near-infrared region, good biocompatibility, and enhanced two-photon excited fluorescence for two-photon bio-imaging is summarized. In addition, several possible future directions in this field are also discussed (146 references). PMID:28772584

  16. Lighting the Way to See Inside Two-Photon Absorption Materials: Structure–Property Relationship and Biological Imaging

    Directory of Open Access Journals (Sweden)

    Qiong Zhang

    2017-02-01

    properties. The metal ions, including transition metals and lanthanides, can serve as an important part of the structure to control the intramolecular charge-transfer process that drives the 2PA process. As templates, transition metal ions can assemble simple to more sophisticated ligands in a variety of multipolar arrangements resulting in interesting and tailorable electronic and optical properties, depending on the nature of the metal center and the energetics of the metal-ligand interactions, such as intraligand charge-transfer (ILCT and metal-ligand charge-transfer (MLCT processes. Lanthanide complexes are attractive for a number of reasons: (i their visible emissions are quite long-lived; (ii their absorption and emission can be tuned with the aid of appropriate photoactive ligands; (iii the accessible energy-transfer path between the photo-active ligands and the lanthanide ion can facilitate efficient lanthanide-based 2PA properties. Thus, the above materials with excellent 2PA properties should be applied in two-photon applications, especially two-photon fluorescence microscopy (TPFM and related emission-based applications. Furthermore, the progress of research into the use of those new 2PA materials with moderate 2PA cross section in the near-infrared region, good Materials 2017, 10, 223 2 of 37 biocompatibility, and enhanced two-photon excited fluorescence for two-photon bio-imaging is summarized. In addition, several possible future directions in this field are also discussed (146 references.

  17. Lighting the Way to See Inside Two-Photon Absorption Materials: Structure-Property Relationship and Biological Imaging.

    Science.gov (United States)

    Zhang, Qiong; Tian, Xiaohe; Zhou, Hongping; Wu, Jieying; Tian, Yupeng

    2017-02-23

    ions, including transition metals and lanthanides, can serve as an important part of the structure to control the intramolecular charge-transfer process that drives the 2PA process. As templates, transition metal ions can assemble simple to more sophisticated ligands in a variety of multipolar arrangements resulting in interesting and tailorable electronic and optical properties, depending on the nature of the metal center and the energetics of the metal-ligand interactions, such as intraligand charge-transfer (ILCT) and metal-ligand charge-transfer (MLCT) processes. Lanthanide complexes are attractive for a number of reasons: (i) their visible emissions are quite long-lived; (ii) their absorption and emission can be tuned with the aid of appropriate photoactive ligands; (iii) the accessible energy-transfer path between the photo-active ligands and the lanthanide ion can facilitate efficient lanthanide-based 2PA properties. Thus, the above materials with excellent 2PA properties should be applied in two-photon applications, especially two-photon fluorescence microscopy (TPFM) and related emission-based applications. Furthermore, the progress of research into the use of those new 2PA materials with moderate 2PA cross section in the near-infrared region, good Materials 2017, 10, 223 2 of 37 biocompatibility, and enhanced two-photon excited fluorescence for two-photon bio-imaging is summarized. In addition, several possible future directions in this field are also discussed (146 references).

  18. Adaptive optics two-photon excited fluorescence lifetime imaging ophthalmoscopy of exogenous fluorophores in mice.

    Science.gov (United States)

    Feeks, James A; Hunter, Jennifer J

    2017-05-01

    In vivo cellular scale fluorescence lifetime imaging of the mouse retina has the potential to be a sensitive marker of retinal cell health. In this study, we demonstrate fluorescence lifetime imaging of extrinsic fluorophores using adaptive optics fluorescence lifetime imaging ophthalmoscopy (AOFLIO). We recorded AOFLIO images of inner retinal cells labeled with enhanced green fluorescent protein (EGFP) and capillaries labeled with fluorescein. We demonstrate that AOFLIO can be used to differentiate spectrally overlapping fluorophores in the retina. With further refinements, AOFLIO could be used to assess retinal health in early stages of degeneration by utilizing lifetime-based sensors or even fluorophores native to the retina.

  19. In vivo high-resolution structural imaging of large arteries in small rodents using two-photon laser scanning microscopy

    Science.gov (United States)

    Megens, Remco T. A.; Reitsma, Sietze; Prinzen, Lenneke; Oude Egbrink, Mirjam G. A.; Engels, Wim; Leenders, Peter J. A.; Brunenberg, Ellen J. L.; Reesink, Koen D.; Janssen, Ben J. A.; Ter Haar Romeny, Bart M.; Slaaf, Dick W.; van Zandvoort, Marc A. M. J.

    2010-01-01

    In vivo (molecular) imaging of the vessel wall of large arteries at subcellular resolution is crucial for unraveling vascular pathophysiology. We previously showed the applicability of two-photon laser scanning microscopy (TPLSM) in mounted arteries ex vivo. However, in vivo TPLSM has thus far suffered from in-frame and between-frame motion artifacts due to arterial movement with cardiac and respiratory activity. Now, motion artifacts are suppressed by accelerated image acquisition triggered on cardiac and respiratory activity. In vivo TPLSM is performed on rat renal and mouse carotid arteries, both surgically exposed and labeled fluorescently (cell nuclei, elastin, and collagen). The use of short acquisition times consistently limit in-frame motion artifacts. Additionally, triggered imaging reduces between-frame artifacts. Indeed, structures in the vessel wall (cell nuclei, elastic laminae) can be imaged at subcellular resolution. In mechanically damaged carotid arteries, even the subendothelial collagen sheet (~1 μm) is visualized using collagen-targeted quantum dots. We demonstrate stable in vivo imaging of large arteries at subcellular resolution using TPLSM triggered on cardiac and respiratory cycles. This creates great opportunities for studying (diseased) arteries in vivo or immediate validation of in vivo molecular imaging techniques such as magnetic resonance imaging (MRI), ultrasound, and positron emission tomography (PET).

  20. 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.

  1. A tale of two photons: radioluminescence and its application in molecular imaging

    Science.gov (United States)

    Pratx, Guillem

    2017-02-01

    Optical and ionizing radiation are two physical ways in which we can probe the living world. Until recently, these forms of radiation were used in distinct imaging and therapeutic applications—radiation therapy, photodynamic therapy, X-ray imaging, and diffuse optical tomography, to name a few. It has now been recognized that physical phenomena in which ionizing radiation and light are inherently coupled may provide powerful new capabilities for imaging and treating diseases. This presentation will review the physics and applications of radioluminescence, with a particular focus on molecular imaging. One such method, X-ray luminescence computed tomography (XLCT), uses narrow kilovolt X-ray beams to stimulate optical emissions from biologically targeted radioluminescent nanoparticles, thus providing high-resolution images even deep in tissue. A different phenomenon, Cherenkov luminescence, can also be harnessed to localize radiopharmaceuticals in vivo, allowing surgeons to visualize the molecular status of the tissues they are resecting. Recent progress towards routine implantation of these methods will be reviewed and sources of endogenous radioluminescence signal will be discussed.

  2. An optimized two-photon method for in vivo lung imaging reveals intimate cell collaborations during infection

    Science.gov (United States)

    Fiole, Daniel; Deman, Pierre; Trescos, Yannick; Douady, Julien; Tournier, Jean-Nicolas

    2013-02-01

    Lung tissue motion arising from breathing and heart beating has been described as the largest annoyance of in vivo imaging. Consequently, infected lung tissue has never been imaged in vivo thus far, and little is known concerning the kinetics of the mucosal immune system at the cellular level. We have developed an optimized post-processing strategy to overcome tissue motion, based upon two-photon and second harmonic generation (SHG) microscopy. In contrast to previously published data, we have freed the lung parenchyma from any strain and depression in order to maintain the lungs under optimal physiological parameters. Excitation beams swept the sample throughout normal breathing and heart movements, allowing the collection of many images. Given that tissue motion is unpredictably, it was essential to sort images of interest. This step was enhanced by using SHG signal from collagen as a reference for sampling and realignment phases. A normalized cross-correlation criterion was used between a manually chosen reference image and rigid transformations of all others. Using CX3CR1+/gfp mice this process allowed the collection of high resolution images of pulmonary dendritic cells (DCs) interacting with Bacillus anthracis spores, a Gram-positive bacteria responsible for anthrax disease. We imaged lung tissue for up to one hour, without interrupting normal lung physiology. Interestingly, our data revealed unexpected interactions between DCs and macrophages, two specialized phagocytes. These contacts may participate in a better coordinate immune response. Our results not only demonstrate the phagocytizing task of lung DCs but also infer a cooperative role of alveolar macrophages and DCs.

  3. Calcium imaging of inner ear hair cells within the cochlear epithelium of mice using two-photon microscopy

    Science.gov (United States)

    Yuan, Tao; Gao, Simon S.; Saggau, Peter; Oghalai, John S.

    2010-01-01

    Mice are an excellent model for studying mammalian hearing and transgenic mouse models of human hearing, loss are commonly available. However, the mouse cochlea is substantially smaller than other animal models routinely used to study cochlear physiology. This makes study of their hair cells difficult. We develop a novel methodology to optically image calcium within living hair cells left undisturbed within the excised mouse cochlea. Fresh cochleae are harvested, left intact within their otic capsule bone, and fixed in a recording chamber. The bone overlying the cochlear epithelium is opened and Reissner's membrane is incised. A fluorescent calcium indicator is applied to the preparation. A custom-built upright two-photon microscope was used to image the preparation using 3-D scanning. We are able to image about one third of a cochlear turn simultaneously, in either the apical or basal regions. Within one hour of animal sacrifice, we find that outer hair cells demonstrate increased fluorescence compared with surrounding supporting cells. This methodology is then used to visualize hair cell calcium changes during mechanotransduction over a region of the epithelium. Because the epithelium is left within the cochlea, dissection trauma is minimized and artifactual changes in hair cell physiology are expected to be reduced.

  4. Calcium rubies: a family of red-emitting functionalizable indicators suitable for two-photon Ca2+ imaging.

    Science.gov (United States)

    Collot, Mayeul; Loukou, Christina; Yakovlev, Aleksey V; Wilms, Christian D; Li, Dongdong; Evrard, Alexis; Zamaleeva, Alsu; Bourdieu, Laurent; Léger, Jean-François; Ropert, Nicole; Eilers, Jens; Oheim, Martin; Feltz, Anne; Mallet, Jean-Maurice

    2012-09-12

    We designed Calcium Rubies, a family of functionalizable BAPTA-based red-fluorescent calcium (Ca(2+)) indicators as new tools for biological Ca(2+) imaging. The specificity of this Ca(2+)-indicator family is its side arm, attached on the ethylene glycol bridge that allows coupling the indicator to various groups while leaving open the possibility of aromatic substitutions on the BAPTA core for tuning the Ca(2+)-binding affinity. Using this possibility we now synthesize and characterize three different CaRubies with affinities between 3 and 22 μM. Their long excitation and emission wavelengths (peaks at 586/604 nm) allow their use in otherwise challenging multicolor experiments, e.g., when combining Ca(2+) uncaging or optogenetic stimulation with Ca(2+) imaging in cells expressing fluorescent proteins. We illustrate this capacity by the detection of Ca(2+) transients evoked by blue light in cultured astrocytes expressing CatCh, a light-sensitive Ca(2+)-translocating channelrhodopsin linked to yellow fluorescent protein. Using time-correlated single-photon counting, we measured fluorescence lifetimes for all CaRubies and demonstrate a 10-fold increase in the average lifetime upon Ca(2+) chelation. Since only the fluorescence quantum yield but not the absorbance of the CaRubies is Ca(2+)-dependent, calibrated two-photon fluorescence excitation measurements of absolute Ca(2+) concentrations are feasible.

  5. Mercury effects on Thalassiosira weissflogii: Applications of two-photon excitation chlorophyll fluorescence lifetime imaging and flow cytometry

    Energy Technology Data Exchange (ETDEWEB)

    Wu Yun [Division of Life Science, Hong Kong University of Science and Technology (HKUST), Clear Water Bay, Kowloon, Hong Kong (Hong Kong); Zeng Yan; Qu, Jianan Y. [Department of Electronic and Computer Engineering, Hong Kong University of Science and Technology (HKUST), Clear Water Bay, Kowloon, Hong Kong (Hong Kong); Wang Wenxiong, E-mail: wwang@ust.hk [Division of Life Science, Hong Kong University of Science and Technology (HKUST), Clear Water Bay, Kowloon, Hong Kong (Hong Kong)

    2012-04-15

    The toxic effects of inorganic mercury [Hg(II)] and methylmercury (MeHg) on the photosynthesis and population growth in a marine diatom Thalassiosira weissflogii were investigated using two methods: two-photon excitation fluorescence lifetime imaging (FLIM) and flow cytometry (FCM). For photosynthesis, Hg(II) exposure increased the average chlorophyll fluorescence lifetime, whereas such increment was not found under MeHg stress. This may be caused by the inhibitory effect of Hg(II) instead of MeHg on the electron transport chain. For population growth, modeled specific growth rate data showed that the reduction in population growth by Hg(II) mainly resulted from an increased number of injured cells, while the live cells divided at the normal rates. However, MeHg inhibitory effects on population growth were contributed by the reduced division rates of all cells. Furthermore, the cell images and the FCM data reflected the morphological changes of diatom cells under Hg(II)/MeHg exposure vividly and quantitatively. Our results demonstrated that the toxigenicity mechanisms between Hg(II) and MeHg were different in the algal cells.

  6. NEAR-IR TWO PHOTON MICROSCOPY IMAGING OF SILICA NANOPARTICLES FUNCTIONALIZED WITH ISOLATED SENSITIZED Yb(III) CENTERS

    Energy Technology Data Exchange (ETDEWEB)

    Lapadula, Giuseppe; Bourdolle, Adrien; Allouche, Florian; Conley, Matthew P.; Maron, Laurent; Lukens, Wayne W.; Guyot, Yannick; Andraud, Chantal; Brasselet, Sophie; Copé; ret, Christophe; Maury, Olivier; Andersen, Richard A.

    2013-01-12

    Bright nano objects emitting in the near infrared with a maximal cross section of 41.4 x 103 GM (Goppert Mayer), were prepared by implanting ca. 180 4,4 diethylaminostyryl 2,2 bipyridine (DEAS) Yb(III) complexes on the surface of 12 nm silica nanoparticles. The surface complexes ([DEAS Ln SiO2], Ln =Y,Yb) were characterized using IR, solid state NMR, UV Vis, EXAFS spectroscopies in combination with the preparation and characterization of similar molecular analogues by analytical techniques (IR, solution NMR, UV Vis, X ray crystallography) as well as DFT calculations. Starting from the partial dehydroxylation of the silica at 700 C on high vacuum having 0.8 OH.nm 2, the grafting of Ln(N(SiMe3)2)3 generate ≤SiO Ln(N(SiMe3)2)2, which upon thermal step and coordination of the DEAS chromophore yields (≤SiO)3Ln(DEAS). Surface and molecular analogues display similar properties, in terms of DEAS binding constants absorption maxima and luminescence properties (intense emission band assigned to a ligand centered CT fluorescence and life time) in the solid state, consistent with the molecular nature of the surface species. The densely functionalized nanoparticles can be dispersed via ultra-sonication in small ca. 15-20 nm aggregates (1 to 6 elementary particles) that were detected using two photon microscopy imaging at 720 nm excitation, making them promising nano objects for bio imaging.

  7. Silole-Based Red Fluorescent Organic Dots for Bright Two-Photon Fluorescence In vitro Cell and In vivo Blood Vessel Imaging.

    Science.gov (United States)

    Chen, Bin; Feng, Guangxue; He, Bairong; Goh, Chiching; Xu, Shidang; Ramos-Ortiz, Gabriel; Aparicio-Ixta, Laura; Zhou, Jian; Ng, Laiguan; Zhao, Zujin; Liu, Bin; Tang, Ben Zhong

    2016-02-10

    Robust luminescent dyes with efficient two-photon fluorescence are highly desirable for biological imaging applications, but those suitable for organic dots fabrication are still rare because of aggregation-caused quenching. In this work, a red fluorescent silole, 2,5-bis[5-(dimesitylboranyl)thiophen-2-yl]-1-methyl-1,3,4-triphenylsilole ((MesB)2 DTTPS), is synthesized and characterized. (MesB)2 DTTPS exhibits enhanced fluorescence efficiency in nanoaggregates, indicative of aggregation-enhanced emission (AEE). The organic dots fabricated by encapsulating (MesB)2 DTTPS within lipid-PEG show red fluorescence peaking at 598 nm and a high fluorescence quantum yield of 32%. Upon excitation at 820 nm, the dots show a large two-photon absorption cross section of 3.43 × 10(5) GM, which yields a two-photon action cross section of 1.09 × 10(5) GM. These (MesB)2 DTTPS dots show good biocompatibility and are successfully applied to one-photon and two-photon fluorescence imaging of MCF-7 cells and two-photon in vivo visualization of the blood vascular of mouse muscle in a high-contrast and noninvasive manner. Moreover, the 3D blood vasculature located at the mouse ear skin with a depth of over 100 μm can also be visualized clearly, providing the spatiotemporal information about the whole blood vascular network. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. 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.

  9. Imaging of goblet cells as a marker for intestinal metaplasia of the stomach by one-photon and two-photon fluorescence endomicroscopy

    Science.gov (United States)

    Bao, Hongchun; Boussioutas, Alex; Reynolds, Jeremy; Russell, Sarah; Gu, Min

    2009-11-01

    Goblet cells are a requirement for the diagnosis of intestinal metaplasia of the stomach. The gastric mucosa is lined by a monolayer of columnar epithelium with some specialization at the crypts, but there are no goblet cells in normal gastric epithelium. The appearance of goblet cells in gastric epithelium is an indicator of potential malignant progression toward adenocarcinoma. Therefore, in vivo three-dimensional imaging of goblet cells is essential for diagnoses of a premalignant stage of gastric cancers called intestinal metaplasia. We used mouse intestine, which has goblet cells, as a model of intestinal metaplasia. One-photon confocal fluorescence endomicroscopy and two-photon fluorescence endomicroscopy are employed for 3-D imaging of goblet cells. The penetration depth, the sectioning ability, and the photobleaching information of imaging are demonstrated. Both endomicroscopy techniques can three-dimensionally observe goblet cells in mouse large intestine and achieve an imaging depth of 176 μm. The two-photon fluorescence endomicroscopy shows higher resolution and contrast of the imaging sections at each depth. In addition, two-photon fluorescence endomicroscopy also has advantages of sectioning ability and less photobleaching. These results prove that two-photon fluorescence endomicroscopy is advantageous in diagnoses of a premalignant stage of gastric cancers.

  10. Two-photon photoemission spectroscopy of image potential states on Ag(001): Momentum-dependent lifetimes and quantum beats

    Energy Technology Data Exchange (ETDEWEB)

    Kiel, Mario; Duncker, Klaus; Widdra, Wolf [Martin-Luther Universitaet Halle-Wittenberg, Halle (Germany)

    2009-07-01

    Using a novel all-fiber based oscillator/amplifier system operating at a repetition rate of 1.5 MHz which pumps two noncollinear optical parametric amplifiers with tunable pulses (<30 fs) in the range of 500-950 nm, the n=1, 2 and 3 image potential states on Ag(001) have been investigated by time and momentum-resolved two-color two-photon photoemission spectroscopy (2PPE). Lifetimes of 55, 165 and 380 fs at k {sub parallel} =0 have been determined for these states, respectively, similar to previous work. The lifetime of the n=1 state reveals a quadratic momentum-dependence of the decay rate. This leads to a shortening to 26 fs at k {sub parallel} =0.4 A{sup -1} which can be described by a linear 34 meV/eV decay rate increase with excess kinetic energy which is comparable to results on Cu(001). It clarifies the controversy regarding the existence of additional decay channels on Ag(001) vs. Cu(001) derived from intrinsic linewidth measurements. Quantum beat spectroscopy is used to determine the energy difference between the n=3 and 4 states to 35 meV based on the 2PPE signal oscillation with a periodicity of 118 fs.

  11. Multifunctional superparamagnetic nanoshells: combining two-photon luminescence imaging, surface-enhanced Raman scattering and magnetic separation

    Science.gov (United States)

    Jin, Xiulong; Li, Haiyan; Wang, Shanshan; Kong, Ni; Xu, Hong; Fu, Qihua; Gu, Hongchen; Ye, Jian

    2014-11-01

    With the increasing need for multi-purpose analysis in the biomedical field, traditional single diagnosis methods cannot meet the requirements. Therefore new multifunctional technologies and materials for the integration of sample collection, sensing and imaging are in great demand. Core-shell nanoparticles offer a unique platform to combine multifunctions in a single particle. In this work, we have constructed a novel type of core-shell superparamagnetic nanoshell (Fe3O4@SiO2@Au), composed of a Fe3O4 cluster core, a thin Au shell and a SiO2 layer in between. The obtained multifunctional nanoparticles combine the magnetic properties and plasmonic optical properties effectively, which were well investigated by a number of experimental characterization methods and theoretical simulations. We have demonstrated that Fe3O4@SiO2@Au nanoparticles can be utilized for two-photon luminescence (TPL) imaging, near-infrared surface-enhanced Raman scattering (NIR SERS) and cell collection by magnetic separation. The TPL intensity could be further greatly enhanced through the plasmon coupling effect in the self-assembled nanoparticle chains, which were triggered by an external magnetic field. In addition, Fe3O4@SiO2@Au nanoparticles may have great potential applications such as enhanced magnetic resonance imaging (MRI) and photo-thermotherapy. Successful combination of multifunctions including magnetic response, biosensing and bioimaging in single nanoparticles allows further manipulation, real-time tracking, and intracellular molecule analysis of live cells at a single-cell level.With the increasing need for multi-purpose analysis in the biomedical field, traditional single diagnosis methods cannot meet the requirements. Therefore new multifunctional technologies and materials for the integration of sample collection, sensing and imaging are in great demand. Core-shell nanoparticles offer a unique platform to combine multifunctions in a single particle. In this work, we have

  12. A new fluorescent probe with a large turn-on signal for imaging nitroreductase in tumor cells and tissues by two-photon microscopy.

    Science.gov (United States)

    Liu, Zhan-Rong; Tang, Yonghe; Xu, An; Lin, Weiying

    2017-03-15

    Hypoxia is the important characteristic of solid tumors, and it may cause the bioactivity of nitroreductase (NTR) to display an elevated level. Hence, the development of effective monitoring methods of NTR in living systems is of great importance for detecting the occurrence and progress of tumors. Toward this goal, a novel two-photon fluorescence turn-on NTR probe GCTPOC-HY, based on the two-photon platform GCTPOC and the NTR recognition site p-nitrobenzyl ether, is designed and synthesized. The probe GCTPOC-HY exhibits eminent properties such as high sensitivity and selectivity, highly stable photo-stability, and low cytotoxicity. Besides, the probe responds to 1.5μg/mL NTR with a 130-fold fluorescence enhancement, which is larger than the reported two-photon fluorescent NTR probes. Moreover, the probe GCTPOC-HY is suitable for fluorescence imaging of NTR in living cells by one- and two-photon modes. Importantly, the probe GCTPOC-HY is successfully applied to monitor NTR in the tumor tissues with a significant fluorescence signal and a penetration depth of 70µm by using two-photon microscopy. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Highly selective red- and green-emitting two-photon fluorescent probes for cysteine detection and their bio-imaging in living cells.

    Science.gov (United States)

    Yang, Zhiguang; Zhao, Ning; Sun, Yuming; Miao, Fang; Liu, Yong; Liu, Xin; Zhang, Yuanhong; Ai, Wentao; Song, Guofen; Shen, Xiaoyuan; Yu, Xiaoqiang; Sun, Jingzhi; Wong, Wai-Yeung

    2012-04-07

    Two highly selective two-photon fluorescent probes for cysteine over homocysteine, N-acetyl-L-cysteine, dithiothreitol, glutathione and other amino acids, and their fluorescent imaging in living cells have been shown. This journal is © The Royal Society of Chemistry 2012

  14. Development of image-guided targeted two-photon PDT for the treatment of head and neck cancers

    Science.gov (United States)

    Spangler, Charles W.; Starkey, Jean R.; Liang, Bo; Fedorka, Sara; Yang, Hao; Jiang, Huabei

    2014-03-01

    There has been significant effort over the past two decades in the treatment of malignancies of epithelial origin, including some of the most devastating of cancers, such as colorectal cancer (CRC), squamous call carcinoma of the head and neck (HNSCC), and carcinomas of the pancreas, lungs, (both Small Cell and Non-Small Cell), renal cell, prostate, bladder and breast. Recurring, refractory HNSCC is a particularly difficult cancer to treat once the tumors recur due to mutations that are resistant to repeat chemotherapy and radiation. In addition, repeat surgery is often difficult due to the requirement of significant surgical margins that may not be possible due to the attending potential functional deficits (e.g., salivary glands, nerves and major blood vessels in confined areas). In this study FaDu HNSCC xenograft tumors in SCID mice were imaged, and "optical", as opposed to "surgical" margins defined for the tumor being treated. The subsequent two-photon treatment irradiation was computer-controlled to carry out the tumor treatment by rastering the laser beam throughout the tumor volume plus the defined optical margins simultaneously. In our initial studies, up to 85% regression in tumor volume was observed in 5 days post PDT, with complete tumor regression in 18 days. No re-growth was observed up to 41 days post-PDT, with little or no scarring and complete hair re-growth. However, competition between imaging and PDT moieties was also observed in some mouse models, possibly favoring tumor re-growth. Strategies to selectively optimize the PDT effect will be discussed.

  15. Direct Vpr-Vpr Interaction in Cells monitored by two Photon Fluorescence Correlation Spectroscopy and Fluorescence Lifetime Imaging

    Directory of Open Access Journals (Sweden)

    Mély Yves

    2008-09-01

    Full Text Available Abstract Background The human immunodeficiency virus type 1 (HIV-1 encodes several regulatory proteins, notably Vpr which influences the survival of the infected cells by causing a G2/M arrest and apoptosis. Such an important role of Vpr in HIV-1 disease progression has fuelled a large number of studies, from its 3D structure to the characterization of specific cellular partners. However, no direct imaging and quantification of Vpr-Vpr interaction in living cells has yet been reported. To address this issue, eGFP- and mCherry proteins were tagged by Vpr, expressed in HeLa cells and their interaction was studied by two photon fluorescence lifetime imaging microscopy and fluorescence correlation spectroscopy. Results Results show that Vpr forms homo-oligomers at or close to the nuclear envelope. Moreover, Vpr dimers and trimers were found in the cytoplasm and in the nucleus. Point mutations in the three α helices of Vpr drastically impaired Vpr oligomerization and localization at the nuclear envelope while point mutations outside the helical regions had no effect. Theoretical structures of Vpr mutants reveal that mutations within the α-helices could perturb the leucine zipper like motifs. The ΔQ44 mutation has the most drastic effect since it likely disrupts the second helix. Finally, all Vpr point mutants caused cell apoptosis suggesting that Vpr-mediated apoptosis functions independently from Vpr oligomerization. Conclusion We report that Vpr oligomerization in HeLa cells relies on the hydrophobic core formed by the three α helices. This oligomerization is required for Vpr localization at the nuclear envelope but not for Vpr-mediated apoptosis.

  16. Two-Photon Na+ Imaging Reports Somatically Evoked Action Potentials in Rat Olfactory Bulb Mitral and Granule Cell Neurites.

    Science.gov (United States)

    Ona-Jodar, Tiffany; Gerkau, Niklas J; Sara Aghvami, S; Rose, Christine R; Egger, Veronica

    2017-01-01

    Dendrodendritic synaptic interactions are a hallmark of neuronal processing in the vertebrate olfactory bulb. Many classes of olfactory bulb neurons including the principal mitral cells (MCs) and the axonless granule cells (GCs) dispose of highly efficient propagation of action potentials (AP) within their dendrites, from where they can release transmitter onto each other. So far, backpropagation in GC dendrites has been investigated indirectly via Ca2+ imaging. Here, we used two-photon Na+ imaging to directly report opening of voltage-gated sodium channels due to AP propagation in both cell types. To this end, neurons in acute slices from juvenile rat bulbs were filled with 1 mM SBFI via whole-cell patch-clamp. Calibration of SBFI signals revealed that a change in fluorescence ΔF/F by 10% corresponded to a Δ[Na+]i of ∼22 mM. We then imaged proximal axon segments of MCs during somatically evoked APs (sAP). While single sAPs were detectable in ∼50% of axons, trains of 20 sAPs at 50 Hz always resulted in substantial ΔF/F of ∼15% (∼33 mM Δ[Na+]i). ΔF/F was significantly larger for 80 Hz vs. 50 Hz trains, and decayed with half-durations τ1/2 ∼0.6 s for both frequencies. In MC lateral dendrites, AP trains yielded small ΔF/F of ∼3% (∼7 mM Δ[Na+]i). In GC apical dendrites and adjacent spines, single sAPs were not detectable. Trains resulted in an average dendritic ΔF/F of 7% (16 mM Δ[Na+]i) with τ1/2 ∼1 s, similar for 50 and 80 Hz. Na+ transients were indistinguishable between large GC spines and their adjacent dendrites. Cell-wise analysis revealed two classes of GCs with the first showing a decrease in ΔF/F along the dendrite with distance from the soma and the second an increase. These classes clustered with morphological parameters. Simulations of Δ[Na+]i replicated these behaviors via negative and positive gradients in Na+ current density, assuming faithful AP backpropagation. Such specializations of dendritic excitability might confer

  17. High speed two-photon imaging of calcium dynamics in dendritic spines: consequences for spine calcium kinetics and buffer capacity.

    Directory of Open Access Journals (Sweden)

    L Niels Cornelisse

    Full Text Available Rapid calcium concentration changes in postsynaptic structures are crucial for synaptic plasticity. Thus far, the determinants of postsynaptic calcium dynamics have been studied predominantly based on the decay kinetics of calcium transients. Calcium rise times in spines in response to single action potentials (AP are almost never measured due to technical limitations, but they could be crucial for synaptic plasticity. With high-speed, precisely-targeted, two-photon point imaging we measured both calcium rise and decay kinetics in spines and secondary dendrites in neocortical pyramidal neurons. We found that both rise and decay kinetics of changes in calcium-indicator fluorescence are about twice as fast in spines. During AP trains, spine calcium changes follow each AP, but not in dendrites. Apart from the higher surface-to-volume ratio (SVR, we observed that neocortical dendritic spines have a markedly smaller endogenous buffer capacity with respect to their parental dendrites. Calcium influx time course and calcium extrusion rate were both in the same range for spines and dendrites when fitted with a dynamic multi-compartment model that included calcium binding kinetics and diffusion. In a subsequent analysis we used this model to investigate which parameters are critical determinants in spine calcium dynamics. The model confirmed the experimental findings: a higher SVR is not sufficient by itself to explain the faster rise time kinetics in spines, but only when paired with a lower buffer capacity in spines. Simulations at zero calcium-dye conditions show that calmodulin is more efficiently activated in spines, which indicates that spine morphology and buffering conditions in neocortical spines favor synaptic plasticity.

  18. High speed two-photon imaging of calcium dynamics in dendritic spines: consequences for spine calcium kinetics and buffer capacity.

    Science.gov (United States)

    Cornelisse, L Niels; van Elburg, Ronald A J; Meredith, Rhiannon M; Yuste, Rafael; Mansvelder, Huibert D

    2007-10-24

    Rapid calcium concentration changes in postsynaptic structures are crucial for synaptic plasticity. Thus far, the determinants of postsynaptic calcium dynamics have been studied predominantly based on the decay kinetics of calcium transients. Calcium rise times in spines in response to single action potentials (AP) are almost never measured due to technical limitations, but they could be crucial for synaptic plasticity. With high-speed, precisely-targeted, two-photon point imaging we measured both calcium rise and decay kinetics in spines and secondary dendrites in neocortical pyramidal neurons. We found that both rise and decay kinetics of changes in calcium-indicator fluorescence are about twice as fast in spines. During AP trains, spine calcium changes follow each AP, but not in dendrites. Apart from the higher surface-to-volume ratio (SVR), we observed that neocortical dendritic spines have a markedly smaller endogenous buffer capacity with respect to their parental dendrites. Calcium influx time course and calcium extrusion rate were both in the same range for spines and dendrites when fitted with a dynamic multi-compartment model that included calcium binding kinetics and diffusion. In a subsequent analysis we used this model to investigate which parameters are critical determinants in spine calcium dynamics. The model confirmed the experimental findings: a higher SVR is not sufficient by itself to explain the faster rise time kinetics in spines, but only when paired with a lower buffer capacity in spines. Simulations at zero calcium-dye conditions show that calmodulin is more efficiently activated in spines, which indicates that spine morphology and buffering conditions in neocortical spines favor synaptic plasticity.

  19. Asante Calcium Green and Asante Calcium Red--novel calcium indicators for two-photon fluorescence lifetime imaging.

    Directory of Open Access Journals (Sweden)

    Karolina Jahn

    Full Text Available For a comprehensive understanding of cellular processes and potential dysfunctions therein, an analysis of the ubiquitous intracellular second messenger calcium is of particular interest. This study examined the suitability of the novel Ca2+-sensitive fluorescent dyes Asante Calcium Red (ACR and Asante Calcium Green (ACG for two-photon (2P-excited time-resolved fluorescence measurements. Both dyes displayed sufficient 2P fluorescence excitation in a range of 720-900 nm. In vitro, ACR and ACG exhibited a biexponential fluorescence decay behavior and the two decay time components in the ns-range could be attributed to the Ca(2+-free and Ca(2+-bound dye species. The amplitude-weighted average fluorescence decay time changed in a Ca(2+-dependent way, unraveling in vitro dissociation constants K(D of 114 nM and 15 nM for ACR and ACG, respectively. In the presence of bovine serum albumin, the absorption and steady-state fluorescence behavior of ACR was altered and its biexponential fluorescence decay showed about 5-times longer decay time components indicating dye-protein interactions. Since no ester derivative of ACG was commercially available, only ACR was evaluated for 2P-excited fluorescence lifetime imaging microscopy (2P-FLIM in living cells of American cockroach salivary glands. In living cells, ACR also exhibited a biexponential fluorescence decay with clearly resolvable short (0.56 ns and long (2.44 ns decay time components attributable to the Ca(2+-free and Ca(2+-bound ACR species. From the amplitude-weighted average fluorescence decay times, an in situ K(D of 180 nM was determined. Thus, quantitative [Ca(2+]i recordings were realized, unraveling a reversible dopamine-induced [Ca(2+]i elevation from 21 nM to 590 nM in salivary duct cells. It was concluded that ACR is a promising new Ca(2+ indicator dye for 2P-FLIM recordings applicable in diverse biological systems.

  20. Biocompatible photoresistant far-red emitting, fluorescent polymer probes, with near-infrared two-photon absorption, for living cell and zebrafish embryo imaging.

    Science.gov (United States)

    Adjili, Salim; Favier, Arnaud; Fargier, Guillaume; Thomas, Audrey; Massin, Julien; Monier, Karine; Favard, Cyril; Vanbelle, Christophe; Bruneau, Sylvia; Peyriéras, Nadine; Andraud, Chantal; Muriaux, Delphine; Charreyre, Marie-Thérèse

    2015-04-01

    Exogenous probes with far-red or near-infrared (NIR) two-photon absorption and fluorescence emission are highly desirable for deep tissue imaging while limiting autofluorescence. However, molecular probes exhibiting such properties are often hydrophobic. As an attractive alternative, we synthesized water-soluble polymer probes carrying multiple far-red fluorophores and demonstrated here their potential for live cell and zebrafish embryo imaging. First, at concentrations up to 10 μm, these polymer probes were not cytotoxic. They could efficiently label living HeLa cells, T lymphocytes and neurons at an optimal concentration of 0.5 μm. Moreover, they exhibited a high resistance to photobleaching in usual microscopy conditions. In addition, these polymer probes could be successfully used for in toto labeling and in vivo two-photon microscopy imaging of developing zebrafish embryos, with remarkable properties in terms of biocompatibility, internalization, diffusion, stability and wavelength emission range. The near-infrared two-photon absorption peak at 910 nm is particularly interesting since it does not excite the zebrafish endogenous fluorescence and is likely to enable long-term time-lapse imaging with limited photodamage. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. 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.

  2. In situ microspatial imaging using two-photon and confocal laser scanning microscopy of bacteria and extracellular polymeric secretions (EPS) within marine stromatolites.

    Science.gov (United States)

    Kawaguchi, Tomohiro; Decho, Alan W

    2002-03-01

    The combination of a hydrophilic embedding resin, Nanoplast, with fluorescent probes, and subsequent imaging using two-photon and confocal laser scanning microscopy (2P-LSM and CLSM) has allowed in imaging of the in situ microspatial arrangements of microbial cells and their extracellular polymeric secretion (EPS) within marine stromatolites. Optical sectioning by 2P-LSM and CLSM allowed imaging of endolithic cyanobacteria cells, Solentia sp., seen within carbonate sand grains. 2P-LSM allowed very clear imaging with a high resolution of bacteria using DAPI, which normally require UV excitation and reduced photo-bleaching of fluorescent probes.

  3. Deep two-photon microscopic imaging through brain tissue using the second singlet state from fluorescent agent chlorophyll α in spinach leaf.

    Science.gov (United States)

    Shi, Lingyan; Rodríguez-Contreras, Adrián; Budansky, Yury; Pu, Yang; Nguyen, Thien An; Alfano, Robert R

    2014-06-01

    Two-photon (2P) excitation of the second singlet (S₂) state was studied to achieve deep optical microscopic imaging in brain tissue when both the excitation (800 nm) and emission (685 nm) wavelengths lie in the "tissue optical window" (650 to 950 nm). S₂ state technique was used to investigate chlorophyll α (Chl α) fluorescence inside a spinach leaf under a thick layer of freshly sliced rat brain tissue in combination with 2P microscopic imaging. Strong emission at the peak wavelength of 685 nm under the 2P S₂ state of Chl α enabled the imaging depth up to 450 μm through rat brain tissue.

  4. Deep two-photon microscopic imaging through brain tissue using the second singlet state from fluorescent agent chlorophyll α in spinach leaf

    Science.gov (United States)

    Shi, Lingyan; Rodríguez-Contreras, Adrián; Budansky, Yury; Pu, Yang; An Nguyen, Thien; Alfano, Robert R.

    2014-06-01

    Two-photon (2P) excitation of the second singlet (S) state was studied to achieve deep optical microscopic imaging in brain tissue when both the excitation (800 nm) and emission (685 nm) wavelengths lie in the "tissue optical window" (650 to 950 nm). S2 state technique was used to investigate chlorophyll α (Chl α) fluorescence inside a spinach leaf under a thick layer of freshly sliced rat brain tissue in combination with 2P microscopic imaging. Strong emission at the peak wavelength of 685 nm under the 2P S state of Chl α enabled the imaging depth up to 450 μm through rat brain tissue.

  5. A series of terpyridine containing flexible amino diethylacetate derivatives with large two-photon action cross-sections for effective mitochondrial imaging in living liver cancerous cells

    Science.gov (United States)

    Jia, Ran; Zhu, Yingying; Hu, Lei; Xiong, Qiru; Zhao, Meng; Zhang, Mingzhu; Tian, Xiaohe

    2018-01-01

    Small molecules possess large two-photon action cross sections (Φσ) are highly demanded for biological purpose. Herein, three novel terpyridine containing flexible amino diethylacetate organic small molecules (A1, A2 and A3) were rationally designed and their photophysical properties were investigated both experimentally and theoretically. The results revealed that the three chromophores possess large Φσ and remarkable Stokes' shift in high polar solvents, which are particularly benefit for further biological imaging application. One chromophore (A1) displayed an effective intracellular uptake against lung cancerous living cells A549. Colocalization studies suggested the internalized subcellular compartment was mitochondria. Consequently, chromophore A1 provides a promising platform to directly monitor mitochondria in living cells under two-photon confocal laser scanning microscopy.

  6. Ruthenium(II) Complex Incorporated UiO-67 Metal-Organic Framework Nanoparticles for Enhanced Two-Photon Fluorescence Imaging and Photodynamic Cancer Therapy.

    Science.gov (United States)

    Chen, Rui; Zhang, Jinfeng; Chelora, Jipsa; Xiong, Yuan; Kershaw, Stephen V; Li, King Fai; Lo, Pik-Kwan; Cheah, Kok Wai; Rogach, Andrey L; Zapien, Juan Antonio; Lee, Chun-Sing

    2017-02-22

    Ruthenium(II) tris(bipyridyl) cationic complex (Ru(bpy) 3 2+ ) incorporated UiO-67 (Universitetet i Oslo) nanoscale metal-organic frameworks (NMOFs) with an average diameter of ∼92 nm were developed as theranostic nanoplatform for in vitro two-photon fluorescence imaging and photodynamic therapy. After incorporation into porous UiO-67 nanoparticles, the quantum yield, luminescence lifetime, and two-photon fluorescence intensity of Ru(bpy) 3 2+ guest molecules were much improved owing to the steric confinement effect of MOF pores. Benefiting from these merits, the as-synthesized nanoparticles managed to be internalized by A549 cells while providing excellent red fluorescence in cytoplasm upon excitation with 880 nm irradiation. Photodynamic therapeutic application of the Ru(bpy) 3 2+ -incorporated UiO-67 NMOFs was investigated in vitro. The Ru(bpy) 3 2+ -incorporated UiO-67 NMOFs exhibited good biocompatibility without irradiation while having good cell-killing rates upon irradiation. In view of these facts, the developed Ru(bpy) 3 2+ -incorporated NMOFs give a new potential pathway to achieve enhanced two-photon fluorescence imaging and photodynamic therapy.

  7. Functional organization of sensory input to the olfactory bulb glomerulus analyzed by two-photon calcium imaging

    Science.gov (United States)

    Wachowiak, Matt; Denk, Winfried; Friedrich, Rainer W.

    2004-06-01

    Glomeruli in the olfactory bulb are anatomically discrete modules receiving input from idiotypic olfactory sensory neurons. To examine the functional organization of sensory inputs to individual glomeruli, we loaded olfactory sensory neurons with a Ca2+ indicator and measured odorant-evoked presynaptic Ca2+ signals within single glomeruli by using two-photon microscopy in anaesthetized mice. Odorants evoked patterns of discrete Ca2+ signals throughout the neuropil of a glomerulus. Across glomeruli, Ca2+ signals occurred with equal probability in all glomerular regions. Within single glomeruli, the pattern of intraglomerular Ca2+ signals was indistinguishable for stimuli of different duration, identity, and concentration. Moreover, the response time course of the signals was similar throughout the glomerulus. Hence, sensory inputs to individual glomeruli are spatially heterogeneous but seem to be functionally indiscriminate. These results support the view of olfactory glomeruli as functional units in representing sensory information.

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

    Science.gov (United States)

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

    2018-01-01

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

  9. Hybrid Theranostic Platform for Second Near-IR Window Light Triggered Selective Two-Photon Imaging and Photothermal Killing of Targeted Melanoma Cells.

    Science.gov (United States)

    Tchounwou, Christine; Sinha, Sudarson Sekhar; Viraka Nellore, Bhanu Priya; Pramanik, Avijit; Kanchanapally, Rajashekhar; Jones, Stacy; Chavva, Suhash Reddy; Ray, Paresh Chandra

    2015-09-23

    Despite advances in the medical field, even in the 21st century cancer is one of the leading causes of death for men and women in the world. Since the second near-infrared (NIR) biological window light between 950 and 1350 nm offers highly efficient tissue penetration, the current article reports the development of hybrid theranostic platform using anti-GD2 antibody attached gold nanoparticle (GNP) conjugated, single-wall carbon nanotube (SWCNT) for second near-IR light triggered selective imaging and efficient photothermal therapy of human melanoma cancer cell. Reported results demonstrate that due to strong plasmon-coupling, two-photon luminescence (TPL) intensity from theranostic GNP attached SWCNT materials is 6 orders of magnitude higher than GNP or SWCNT alone. Experimental and FDTD simulation data indicate that the huge enhancement of TPL intensity is mainly due to strong resonance enhancement coupled with the stronger electric field enhancement. Due to plasmon coupling, the theranostic material serves as a local nanoantennae to enhance the photothermal capability via strong optical energy absorption. Reported data show that theranostic SWCNT can be used for selective two-photon imaging of melanoma UACC903 cell using 1100 nm light. Photothermal killing experiment with 1.0 W/cm(2) 980 nm laser light demonstrates that 100% of melanoma UACC903 cells can be killed using theranostic SWCNT bind melanoma cells after just 8 min of exposure. These results demonstrate that due to plasmon coupling, the theranostic GNP attached SWCNT material serves as a two-photon imaging and photothermal source for cancer cells in biological window II.

  10. Two-photon Imaging of Intracellular Ca2+ Handling and Nitric Oxide Production in Endothelial and Smooth Muscle Cells of an Isolated Rat Aorta.

    Science.gov (United States)

    Endres, Bradley T; Staruschenko, Alexander; Schulte, Marie; Geurts, Aron M; Palygin, Oleg

    2015-06-10

    Calcium is a very important regulator of many physiological processes in vascular tissues. Most endothelial and smooth muscle functions highly depend on changes in intracellular calcium ([Ca(2+)]i) and nitric oxide (NO). In order to understand how [Ca(2+)]i, NO and downstream molecules are handled by a blood vessel in response to vasoconstrictors and vasodilators, we developed a novel technique that applies calcium-labeling (or NO-labeling) dyes with two photon microscopy to measure calcium handling (or NO production) in isolated blood vessels. Described here is a detailed step-by-step procedure that demonstrates how to isolate an aorta from a rat, label calcium or NO within the endothelial or smooth muscle cells, and image calcium transients (or NO production) using a two photon microscope following physiological or pharmacological stimuli. The benefits of using the method are multi-fold: 1) it is possible to simultaneously measure calcium transients in both endothelial cells and smooth muscle cells in response to different stimuli; 2) it allows one to image endothelial cells and smooth muscle cells in their native setting; 3) this method is very sensitive to intracellular calcium or NO changes and generates high resolution images for precise measurements; and 4) described approach can be applied to the measurement of other molecules, such as reactive oxygen species. In summary, application of two photon laser emission microscopy to monitor calcium transients and NO production in the endothelial and smooth muscle cells of an isolated blood vessel has provided high quality quantitative data and promoted our understanding of the mechanisms regulating vascular function.

  11. Water soluble two-photon fluorescent organic probes for long-term imaging of lysosomes in live cells and tumor spheroids.

    Science.gov (United States)

    Kumari, Pratibha; Verma, Sanjay K; Mobin, Shaikh M

    2018-01-11

    The morphological alteration of lysosomes is a powerful indicator of various pathological disorders. In this regard, we have designed and synthesized a new water soluble fluorescent Schiff-base ligand (L-lyso) containing two hydroxyl groups. L-lyso exhibits excellent two-photon properties with tracking of lysosomes in live cells as well as in 3D tumor spheroids. Furthermore, it can label lysosomes for more than 3 days. Thus, L-lyso has an edge over the commercially available expensive LysoTracker probes and also over other reported probes in terms of its long-term imaging, water solubility and facile synthesis.

  12. Noninvasive redox and back-scattered light imaging of keratocyte cells in the cornea: two-photon excitation and scanning slit confocal microscopy

    Science.gov (United States)

    Masters, Barry R.

    1995-04-01

    The ability to image and monitor the metabolic activity of keratocytes is important for the investigation of wound healing and repair mechanisms in the cornea. After laser refractive surgery there is activation of the stromal keratocytes in the human cornea. Two-photon excitation laser scanning microscopy was used to monitor the NAD(P)H levels in keratocytes in the cornea. The autofluorescence was confirmed to be mostly of NAD(P)H origin by treatment with cyanide which caused an increase in the fluorescence by a factor of two. We used a real-time scanning slit confocal microscope to image the distribution of keratocytes in the full thickness of the cornea. This microscope has the ability to image the cellular processes as well as the nuclei of the stromal keratocytes. Noninvasive optical imaging may provide a useful tool to investigate keratocyte activation after laser surgery or wound healing.

  13. The spatiotemporal organization of cerebellar network activity resolved by two-photon imaging of multiple single neurons

    Directory of Open Access Journals (Sweden)

    Daniela eGandolfi

    2014-04-01

    Full Text Available In order to investigate the spatiotemporal organization of neuronal activity in local microcircuits, techniques allowing the simultaneous recording from multiple single neurons are required. To this end, we implemented an advanced spatial-light modulator two-photon microscope (SLM-2PM. A critical issue for cerebellar theory is the organization of granular layer activity in the cerebellum, which has been predicted by single-cell recordings and computational models. With SLM-2PM, calcium signals could be recorded from different network elements in acute cerebellar slices including granule cells (GrCs, Purkinje cells (PCsand molecular layer interneurons. By combining WCRs with SLM-2PM, the spike/calcium relationship in GrCs and PCs could be extrapolated toward the detection of single spikes. The SLM-2PM technique made it possible to monitor activity of over tens to hundreds neurons simultaneously. GrC activity depended on the number of spikes in the input mossy fiber bursts. PC and molecular layer interneuron activity paralleled that in the underlying GrC population revealing the spread of activity through the cerebellar cortical network. Moreover, circuit activity was increased by the GABA-A receptor blocker, gabazine, and reduced by the AMPA and NMDA receptor blockers, NBQX and APV. The SLM-2PM analysis of spatiotemporal patterns lent experimental support to the time-window and center-surround organizing principles of the granular layer.

  14. Single pulse two-photon fluorescence lifetime imaging (SP-FLIM) with MHz pixel rate and an all fiber based setup

    Science.gov (United States)

    Eibl, Matthias; Karpf, Sebastian; Hakert, Hubertus; Weng, Daniel; Pfeiffer, Tom; Kolb, Jan Philip; Huber, Robert

    2017-07-01

    Newly developed microscopy methods have the goal to give researches in bio-molecular science a better understanding of processes ongoing on a cellular level. Especially two-photon excited fluorescence (TPEF) microscopy is a readily applied and widespread modality. Compared to one photon fluorescence imaging, it is possible to image not only the surface but also deeper lying structures. Together with fluorescence lifetime imaging (FLIM), which provides information on the chemical composition of a specimen, deeper insights on a molecular level can be gained. However, the need for elaborate light sources for TPEF and speed limitations for FLIM hinder an even wider application. In this contribution, 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 perfectly suited for fiber delivery as typically limiting non-linear effects like self-phase or cross-phase modulation (SPM, XPM) are negligible. Furthermore, compared to the typically applied femtosecond pulses, our longer pulses produce much more fluorescence photons per single shot. In this paper, we show that this 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 our system, we acquired FLIM images of a dye solution with single exponential behavior to assess the accuracy of our lifetime determination and also FLIM images of a plant stem at a pixel rate of 1 MHz to show the speed performance of our single pulse two-photon FLIM (SP-FLIM) system.

  15. A feasible add-on upgrade on a commercial two-photon FLIM microscope for optimal FLIM-FRET imaging of CFP-YFP pairs.

    Science.gov (United States)

    Xu, Lingling; Wang, Liang; Zhang, Zhihong; Huang, Zhen-Li

    2013-05-01

    Fluorescence lifetime imaging microscopy (FLIM) based on time-correlated single photon counting (TCSPC) is a widely used method for fluorescence resonance energy transfer (FRET). Here we report a feasible add-on approach to upgrade a commercial two-photon FLIM microscope into a single-photon FLIM microscope which provides optimal FLIM-FRET imaging of FRET pairs consisting of cyan fluorescent proteins (CFPs) as the donor and yellow fluorescent proteins (YFPs) as the acceptor. The capability of the upgraded system is evaluated and discussed, and the imaging performance of the system is demonstrated using FLIM-FRET experiments with a representative CFP-YFP FRET pair (mCerulean-mCitrine).

  16. Scanless multitarget-matching multiphoton excitation fluorescence microscopy

    Directory of Open Access Journals (Sweden)

    Junpeng Qiu

    2018-03-01

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

  17. 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.

  18. Design of a two-photon fluorescent probe for selective recognition of Au(III) over Au(I) and its application of imaging in vitro and in vivo

    Science.gov (United States)

    Wang, Wenjuan; Huang, Yinliang; Wang, Shumin; Zhou, Yujie; Huang, Wei; Feng, Yan; Zhang, Wan; Yu, Wenxin; Zhou, Qiang; Chen, Man; Fang, Min

    2017-12-01

    A highly selective two-photon fluorescent probe (PyCM-1) for Au3 + was developed with distinct ;turn on; fluorescence response, low detection limit (22 nM) and large two-photon absorption cross-sections (696 GM at 860 nm). Its high selectivity for Au3 + over Au+ was achieved via the modification on the type of coordination atoms in the Schiff base receptor. Co-staining experiments showed that the probe PyCM-1 could co-localize specifically with mitochondria. Moreover, the two-photon confocal fluorescence imaging results demonstrated the probe's capability for visualizing Au3 +in vitro and in vivo.

  19. 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.

  20. Antibody Conjugated, Raman Tagged Hollow Gold-Silver Nanospheres for Specific Targeting and Multimodal Dark-Field/SERS/Two Photon-FLIM Imaging of CD19(+) B Lymphoblasts.

    Science.gov (United States)

    Nagy-Simon, Timea; Tatar, Andra-Sorina; Craciun, Ana-Maria; Vulpoi, Adriana; Jurj, Maria-Ancuta; Florea, Adrian; Tomuleasa, Ciprian; Berindan-Neagoe, Ioana; Astilean, Simion; Boca, Sanda

    2017-06-28

    In this Research Article, we propose a new class of contrast agents for the detection and multimodal imaging of CD19(+) cancer lymphoblasts. The agents are based on NIR responsive hollow gold-silver nanospheres conjugated with antiCD19 monoclonal antibodies and marked with Nile Blue (NB) SERS active molecules (HNS-NB-PEG-antiCD19). Proof of concept experiments on specificity of the complex for the investigated cells was achieved by transmission electron microscopy (TEM). The microspectroscopic investigations via dark field (DF), surface-enhanced Raman spectroscopy (SERS), and two-photon excited fluorescence lifetime imaging microscopy (TPE-FLIM) corroborate with TEM and demonstrate successful and preferential internalization of the antibody-nanocomplex. The combination of the microspectroscopic techniques enables contrast and sensitivity that competes with more invasive and time demanding cell imaging modalities, while depth sectioning images provide real time localization of the nanoparticles in the whole cytoplasm at the entire depth of the cells. Our findings prove that HNS-NB-PEG-antiCD19 represent a promising type of new contrast agents with great possibility of being detected by multiple, non invasive, rapid and accessible microspectroscopic techniques and real applicability for specific targeting of CD19(+) cancer cells. Such versatile nanocomplexes combine in one single platform the detection and imaging of cancer lymphoblasts by DF, SERS, and TPE-FLIM microspectroscopy.

  1. 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.

  2. In-situ characterization of the uncrimping process of arterial collagen fibers using two-photon confocal microscopy and digital image correlation.

    Science.gov (United States)

    Wang, Ruoya; Brewster, Luke P; Gleason, Rudolph L

    2013-10-18

    Uncrimping of collagen fibers in the arterial wall is an integral process in regulating the macro-level mechanical response of arteries. Uncrimping of collagen fibers leads to a gradual, but significant strain-stiffening response of the artery at physiological pressures and prevents overdistention at elevated pressures. In this study, we imaged adventitial collagen fibers from fresh primate arteries using two-photon excitation microscopy while subjecting the arteries to physiological inflation pressures and axial stretches. The imaging focal plane was fixed at a constant radial location in the adventitial wall by adjusting the focal distance as the arteries inflated, allowing for the continuously monitoring of the uncrimping process of a single region of collagen fibers. Digital image correlation was then applied to the sequential images to assess and correlate the local displacements to manual traces of selected reference fibers and their engagements. We found that the collagen fibers of interest became fully engaged at a luminal pressure of 20mmHg, this was then followed by rotation of these fibers as the bulk artery continued to dilate. This technique helps to further the understanding of the uncrimping process of collagen fibers under physiological loads, which can aid in the development of more accurate microstructural constitutive models. © 2013 Elsevier Ltd. All rights reserved.

  3. Opposite reactivity of meningeal versus cortical microvessels to the nitric oxide donor glyceryl trinitrate evaluated in vivo with two-photon imaging.

    Directory of Open Access Journals (Sweden)

    Evgeny Pryazhnikov

    Full Text Available Vascular changes underlying headache in migraine patients induced by Glyceryl trinitrate (GTN were previously studied with various imaging techniques. Despite the long history of medical and experimental use of GTN, its effects on the brain vasculature are still poorly understood presumably due to low spatial resolution of the imaging modalities used so far. We took advantage of the micrometer-scale vertical resolution of two-photon microscopy to differentiate between the vasodynamic effects of GTN on meningeal versus cortical vessels imaged simultaneously in anesthetized rats through either thinned skull or glass-sealed cranial window. Intermediate and small calibre vessels were visualized in vivo by imaging intravascular fluorescent dextran, and detection of blood flow direction allowed identification of individual arterioles and venules. We found that i.p.-injected GTN induced a transient constriction of meningeal arterioles, while their cortical counterparts were, in contrast, dilated. These opposing effects of GTN were restricted to arterioles, whereas the effects on venules were insignificant. Interestingly, the NO synthase inhibitor L-NAME did not affect the diameter of meningeal vessels but induced a constriction of cortical vessels. The different cellular environment in cortex versus meninges as well as distinct vessel wall anatomical features probably play crucial role in the observed phenomena. These findings highlight differential region- and vessel-type-specific effects of GTN on cranial vessels, and may implicate new vascular mechanisms of NO-mediated primary headaches.

  4. Mapping live cell viscosity with an aggregation-induced emission fluorogen by means of two-photon fluorescence lifetime imaging.

    Science.gov (United States)

    Chen, Sijie; Hong, Yuning; Zeng, Yan; Sun, Qiqi; Liu, Yang; Zhao, Engui; Bai, Gongxun; Qu, Jianan; Hao, Jianhua; Tang, Ben Zhong

    2015-03-09

    Intracellular viscosity is a crucial parameter that indicates the functioning of cells. In this work, we demonstrate the utility of TPE-Cy, a cell-permeable dye with aggregation-induced emission (AIE) property, in mapping the viscosity inside live cells. Owing to the AIE characteristics, both the fluorescence intensity and lifetime of this dye are increased along with an increase in viscosity. Fluorescence lifetime imaging of live cells stained with TPE-Cy reveals that the lifetime in lipid droplets is much shorter than that from the general cytoplasmic region. The loose packing of the lipids in a lipid droplet results in low viscosity and thus shorter lifetime of TPE-Cy in this region. It demonstrates that the AIE dye could provide good resolution in intracellular viscosity sensing. This is also the first work in which AIE molecules are applied in fluorescence lifetime imaging and intracellular viscosity sensing. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Label-free imaging of Drosophila in vivo by coherent anti-Stokes Raman scattering and two-photon excitation autofluorescence microscopy

    Science.gov (United States)

    Chien, Cheng-Hao; Chen, Wei-Wen; Wu, June-Tai; Chang, Ta-Chau

    2011-01-01

    Drosophila is one of the most valuable model organisms for studying genetics and developmental biology. The fat body in Drosophila, which is analogous to the liver and adipose tissue in human, stores lipids that act as an energy source during its development. At the early stages of metamorphosis, the fat body remodeling occurs involving the dissociation of the fat body into individual fat cells. Here we introduce a combination of coherent anti-Stokes Raman scattering (CARS) and two-photon excitation autofluorescence (TPE-F) microscopy to achieve label-free imaging of Drosophila in vivo at larval and pupal stages. The strong CARS signal from lipids allows direct imaging of the larval fat body and pupal fat cells. In addition, the use of TPE-F microscopy allows the observation of other internal organs in the larva and autofluorescent globules in fat cells. During the dissociation of the fat body, the findings of the degradation of lipid droplets and an increase in autofluorescent globules indicate the consumption of lipids and the recruitment of proteins in fat cells. Through in vivo imaging and direct monitoring, CARS microscopy may help elucidate how metamorphosis is regulated and study the lipid metabolism in Drosophila.

  6. A Novel Cervical Spinal Cord Window Preparation Allows for Two-Photon Imaging of T-Cell Interactions with the Cervical Spinal Cord Microvasculature during Experimental Autoimmune Encephalomyelitis

    Science.gov (United States)

    Haghayegh Jahromi, Neda; Tardent, Heidi; Enzmann, Gaby; Deutsch, Urban; Kawakami, Naoto; Bittner, Stefan; Vestweber, Dietmar; Zipp, Frauke; Stein, Jens V.; Engelhardt, Britta

    2017-01-01

    T-cell migration across the blood–brain barrier (BBB) is a crucial step in the pathogenesis of experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). Two-photon intravital microscopy (2P-IVM) has been established as a powerful tool to study cell–cell interactions in inflammatory EAE lesions in living animals. In EAE, central nervous system inflammation is strongly pronounced in the spinal cord, an organ in which 2P-IVM imaging is technically very challenging and has been limited to the lumbar spinal cord. Here, we describe a novel spinal cord window preparation allowing to use 2P-IVM to image immune cell interactions with the cervical spinal cord microvascular endothelium during EAE. We describe differences in the angioarchitecture of the cervical spinal cord versus the lumbar spinal cord, which will entail different hemodynamic parameters in these different vascular beds. Using T cells as an example, we demonstrate the suitability of this novel methodology in imaging the post-arrest multistep T-cell extravasation across the cervical spinal cord microvessels. The novel methodology includes an outlook to the analysis of the cellular pathway of T-cell diapedesis across the BBB by establishing visualization of endothelial junctions in this vascular bed. PMID:28443093

  7. Two-photon imaging of cortical surface microvessels reveals a robust redistribution in blood flow after vascular occlusion.

    Directory of Open Access Journals (Sweden)

    Chris B Schaffer

    2006-02-01

    Full Text Available A highly interconnected network of arterioles overlies mammalian cortex to route blood to the cortical mantle. Here we test if this angioarchitecture can ensure that the supply of blood is redistributed after vascular occlusion. We use rodent parietal cortex as a model system and image the flow of red blood cells in individual microvessels. Changes in flow are quantified in response to photothrombotic occlusions to individual pial arterioles as well as to physical occlusions of the middle cerebral artery (MCA, the primary source of blood to this network. We observe that perfusion is rapidly reestablished at the first branch downstream from a photothrombotic occlusion through a reversal in flow in one vessel. More distal downstream arterioles also show reversals in flow. Further, occlusion of the MCA leads to reversals in flow through approximately half of the downstream but distant arterioles. Thus the cortical arteriolar network supports collateral flow that may mitigate the effects of vessel obstruction, as may occur secondary to neurovascular pathology.

  8. A novel technique for the in vivo imaging of autoimmune diabetes development in the pancreas by two-photon microscopy.

    Directory of Open Access Journals (Sweden)

    Ken Coppieters

    Full Text Available Type 1 diabetes (T1D is characterized by the immune-mediated destruction of beta cells in the pancreas. Little is known about the in vivo dynamic interactions between T cells and beta cells or the kinetic behavior of other immune cell subsets in the pancreatic islets. Utilizing multiphoton microscopy we have designed a technique that allows for the real-time visualization of diabetogenic T cells and dendritic cells in pancreatic islets in a live animal, including their interplay with beta cells and the vasculature. Using a custom designed stage, the pancreas was surgically exposed under live conditions so that imaging of islets under intact blood pressure and oxygen supply became possible. We demonstrate here that this approach allows for the tracking of diabetogenic leukocytes as well as vascularization phenotype of islets and accumulation of dendritic cells in islets during diabetes pathogenesis. This technique should be useful in mapping crucial kinetic events in T1D pathogenesis and in testing the impact of immune based interventions on T cell migration, extravasation and islet destruction.

  9. Slow-electron velocity-map imaging study of aniline via resonance-enhanced two-photon ionization method

    Science.gov (United States)

    Qu, Zehua; Qin, Zhengbo; Zheng, Xianfeng; Wang, Hui; Yao, Guanxin; Zhang, Xianyi; Cui, Zhifeng

    2017-02-01

    Slow electron velocity-map imaging (SEVI) of aniline has been investigated via two-color resonant-enhanced two-photo (1 + 1‧) ionization (2C-R2PI) method. A number of vibrational frequencies in the first excited state of neutral (S1) and 2B1 ground electronic state of cation (D0) have been accurately determined. In addition, photoelectron angular distributions (PADs) in the two-step transitions are presented and reveal a near threshold shape resonance in the ionization of aniline. The SEVI spectra taken via various S1 intermediate states provide the detailed vibrational structures of D0 state and directly deduce the accurate adiabatic ionization potential (IP) of 62,271 ± 6 cm- 1. Ab initio calculations excellently reproduce the experimental IP value (Theo. 62,242 cm- 1). For most vibrational modes, good agreement between theoretical and experimental frequencies in the S0 and D0 states of aniline is obtained to aid us to clearly assign vibrational modes. Especially, the vibrational frequencies calculated at the CASSCF level are much better consistent with experimental data than that obtained using the TDDFT and CIS methods.

  10. Two-photon excitation STED microscopy.

    Science.gov (United States)

    Moneron, Gael; Hell, Stefan W

    2009-08-17

    We report sub-diffraction resolution in two-photon excitation (TPE) fluorescence microscopy achieved by merging this technique with stimulated-emission depletion (STED). We demonstrate an easy-to-implement and promising laser combination based on a short-pulse laser source for two-photon excitation and a continuous-wave (CW) laser source for resolution enhancement. Images of fluorescent nanoparticles and the immunostained transcription regulator NF kappaB in mammalian cell nuclei exhibit resolutions of barrier. (c) 2009 Optical Society of America

  11. Wolf equations for two-photon light.

    Science.gov (United States)

    Saleh, Bahaa E A; Teich, Malvin C; Sergienko, Alexander V

    2005-06-10

    The spatiotemporal two-photon probability amplitude that describes light in a two-photon entangled state obeys equations identical to the Wolf equations, which are satisfied by the mutual coherence function for light in any quantum state. Both functions therefore propagate similarly through optical systems. A generalized van Cittert-Zernike theorem explains the predicted enhancement in resolution for entangled-photon microscopy and quantum lithography. The Wolf equations provide a particularly powerful analytical tool for studying three-dimensional imaging and lithography since they describe propagation in continuous inhomogeneous media.

  12. Two-photon and two-photon-assisted slow light.

    Science.gov (United States)

    Bautista, E Sánchez; Cabrera-Granado, E; Weigand, R

    2011-03-01

    We show that light pulses propagating in two-photon absorbing systems may present time delays like slow light produced via coherent population oscillations in one-photon interactions. Two regimes are numerically studied for a simplified two-level system: (a) a light pulse at frequency ω/2 undergoes two-photon absorption (TPA) and is delayed by the absorbing system (two-photon slow light) and (b) a light pulse at frequency ω is delayed in a system prepared by TPA of a light pulse at frequency ω/2 (two-photon-assisted slow light). The study carried out in solutions of dyes and dendrites shows significant delays, low distortion, and good transmission for easily reachable experimental conditions. The working principle can be applied to other media and can be used in telecommunications technology.

  13. Two-photon microscopy imaging of thy1GFP-M transgenic mice: a novel animal model to investigate brain dendritic cell subsets in vivo.

    Directory of Open Access Journals (Sweden)

    Claudia Laperchia

    Full Text Available Transgenic mice expressing fluorescent proteins in specific cell populations are widely used for in vivo brain studies with two-photon fluorescence (TPF microscopy. Mice of the thy1GFP-M line have been engineered for selective expression of green fluorescent protein (GFP in neuronal populations. Here, we report that TPF microscopy reveals, at the brain surface of these mice, also motile non-neuronal GFP+ cells. We have analyzed the behavior of these cells in vivo and characterized in brain sections their immunophenotype.With TPF imaging, motile GFP+ cells were found in the meninges, subarachnoid space and upper cortical layers. The striking feature of these cells was their ability to move across the brain parenchyma, exhibiting evident shape changes during their scanning-like motion. In brain sections, GFP+ cells were immunonegative to antigens recognizing motile cells such as migratory neuroblasts, neuronal and glial precursors, mast cells, and fibroblasts. GFP+ non-neuronal cells exhibited instead the characteristic features and immunophenotype (CD11c and major histocompatibility complex molecule class II immunopositivity of dendritic cells (DCs, and were immunonegative to the microglial marker Iba-1. GFP+ cells were also identified in lymph nodes and blood of thy1GFP-M mice, supporting their identity as DCs. Thus, TPF microscopy has here allowed the visualization for the first time of the motile behavior of brain DCs in situ. The results indicate that the thy1GFP-M mouse line provides a novel animal model for the study of subsets of these professional antigen-presenting cells in the brain. Information on brain DCs is still very limited and imaging in thy1GFP-M mice has a great potential for analyses of DC-neuron interaction in normal and pathological conditions.

  14. Two-Photon Microscopy Imaging of thy1GFP-M Transgenic Mice: A Novel Animal Model to Investigate Brain Dendritic Cell Subsets In Vivo

    Science.gov (United States)

    Laperchia, Claudia; Allegra Mascaro, Anna L.; Sacconi, Leonardo; Andrioli, Anna; Mattè, Alessandro; De Franceschi, Lucia; Grassi-Zucconi, Gigliola; Bentivoglio, Marina; Buffelli, Mario; Pavone, Francesco S.

    2013-01-01

    Transgenic mice expressing fluorescent proteins in specific cell populations are widely used for in vivo brain studies with two-photon fluorescence (TPF) microscopy. Mice of the thy1GFP-M line have been engineered for selective expression of green fluorescent protein (GFP) in neuronal populations. Here, we report that TPF microscopy reveals, at the brain surface of these mice, also motile non-neuronal GFP+ cells. We have analyzed the behavior of these cells in vivo and characterized in brain sections their immunophenotype. With TPF imaging, motile GFP+ cells were found in the meninges, subarachnoid space and upper cortical layers. The striking feature of these cells was their ability to move across the brain parenchyma, exhibiting evident shape changes during their scanning-like motion. In brain sections, GFP+ cells were immunonegative to antigens recognizing motile cells such as migratory neuroblasts, neuronal and glial precursors, mast cells, and fibroblasts. GFP+ non-neuronal cells exhibited instead the characteristic features and immunophenotype (CD11c and major histocompatibility complex molecule class II immunopositivity) of dendritic cells (DCs), and were immunonegative to the microglial marker Iba-1. GFP+ cells were also identified in lymph nodes and blood of thy1GFP-M mice, supporting their identity as DCs. Thus, TPF microscopy has here allowed the visualization for the first time of the motile behavior of brain DCs in situ. The results indicate that the thy1GFP-M mouse line provides a novel animal model for the study of subsets of these professional antigen-presenting cells in the brain. Information on brain DCs is still very limited and imaging in thy1GFP-M mice has a great potential for analyses of DC-neuron interaction in normal and pathological conditions. PMID:23409142

  15. Selective imaging of active pharmaceutical ingredients in powdered blends with common excipients utilizing two-photon excited ultraviolet-fluorescence and ultraviolet-second order nonlinear optical imaging of chiral crystals.

    Science.gov (United States)

    Toth, S J; Madden, J T; Taylor, L S; Marsac, P; Simpson, G J

    2012-07-17

    Second order nonlinear optical imaging of chiral crystals (SONICC) and two-photon excited fluorescence measurements [both autofluorescence and two-photon excited UV-fluorescence (TPE-UVF)] were assessed for the selective detection of APIs relative to common pharmaceutical excipients. Active pharmaceutical ingredients (APIs) compose only a small percentage of most tabulated formulations, yet the API distribution within the tablet can affect drug release and tablet stability. Complementary measurements using either UV-SONICC (266 nm detection) or TPE-UVF were shown to generate signals >50-fold more intense for a model API (griseofulvin) than those produced by common pharmaceutical excipients. The combined product of the measurements produced signals >10(4)-fold greater than the excipients studied. UV-SONICC or TPE-UVF produced greater selectivity than analogous measurements with visible-light detection, attributed to the presence of aromatic moieties within the API exhibiting strong one and two photon absorption at ~266 nm. Complementary SONICC and fluorescence measurements allowed for the sensitive detection of the three-dimensional distribution of tadalafil within a Cialis tablet to a depth of >140 μm.

  16. Two Photon Polymerization of Ormosils

    Science.gov (United States)

    Matei, A.; Zamfirescu, M.; Jipa, F.; Luculescu, C.; Dinescu, M.; Buruiana, E. C.; Buruiana, T.; Sima, L. E.; Petrescu, S. M.

    2010-10-01

    In this work, 3D structures of hybrid polymers—ORMOSILS (organically modified silicates) were produced via Two Photon Polymerization (2PP) of hybrid methacrylates based on silane derivates. Synthetic routes have been used to obtain series of hybrid monomers, their structure and purity being checked by NMR Spectroscopy and Fourier Transform Infrared Spectroscopy. Two photon polymerization method (a relatively new technology which allows fast micro and nano processing of three-dimensional structures with application in medical devices, tissue scaffolds, photonic crystals etc) was used for monomers processing. As laser a Ti: Sapphire laser was used, with 200 fs pulse duration and 2 kHz repetition rate, emitting at 775 nm. A parametric study on the influence of the processing parameters (laser fluence, laser scanning velocity, photo initiator) on the written structures was carried out. The as prepared polymeric scaffolds were tested in mesenchymal stem cells and fibroblasts cell cultures, with the aim of further obtaining bone and dermal grafts. Cells morphology, proliferation, adhesion and alignment were analyzed for different experimental conditions.

  17. 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.

  18. Water-soluble small-molecule probes for RNA based on a two-photon fluorescence "off-on" process: systematic analysis in live cell imaging and understanding of structure-activity relationships.

    Science.gov (United States)

    Li, Hong; Li, Yuncang; Zhang, Huihui; Xu, Guoyong; Zhang, Yuliang; Liu, Xiaohu; Zhou, Hongping; Yang, Xingyuan; Zhang, Xuanjun; Tian, Yupeng

    2017-12-12

    The conveniently obtained water-soluble small-molecule L1-5 were selective for RNA with remarkable "off-on" two-photon fluorescence responses in vitro and in vivo, showing that L1-5 can surprisingly stain mitochondria or nucleoli individually with orange fluorescence under different focal lengths. Compounds L6-8 were prepared, for comparison, to further explore the mechanism of the binding of L1-5 with RNA, which revealed that the synergistic effect of the amino group and the pyridinium cation played a significant role in distinct cell imaging.

  19. Two-photon directed evolution of green fluorescent proteins

    Science.gov (United States)

    Stoltzfus, Caleb R.; Barnett, Lauren M.; Drobizhev, Mikhail; Wicks, Geoffrey; Mikhaylov, Alexander; Hughes, Thomas E.; Rebane, Aleksander

    2015-07-01

    Directed evolution has been used extensively to improve the properties of a variety of fluorescent proteins (FPs). Evolutionary strategies, however, have not yet been used to improve the two-photon absorption (2PA) properties of a fluorescent protein, properties that are important for two-photon imaging in living tissues, including the brain. Here we demonstrate a technique for quantitatively screening the two-photon excited fluorescence (2PEF) efficiency and 2PA cross section of tens of thousands of mutant FPs expressed in E. coli colonies. We use this procedure to move EGFP through three rounds of two-photon directed evolution leading to new variants showing up to a 50% enhancement in peak 2PA cross section and brightness within the near-IR tissue transparency wavelength range.

  20. Reversible Disruption of Neuronal Mitochondria by Ischemic and Traumatic Injury Revealed by Quantitative Two-Photon Imaging in the Neocortex of Anesthetized Mice.

    Science.gov (United States)

    Kislin, Mikhail; Sword, Jeremy; Fomitcheva, Ioulia V; Croom, Deborah; Pryazhnikov, Evgeny; Lihavainen, Eero; Toptunov, Dmytro; Rauvala, Heikki; Ribeiro, Andre S; Khiroug, Leonard; Kirov, Sergei A

    2017-01-11

    Mitochondria play a variety of functional roles in cortical neurons, from metabolic support and neuroprotection to the release of cytokines that trigger apoptosis. In dendrites, mitochondrial structure is closely linked to their function, and fragmentation (fission) of the normally elongated mitochondria indicates loss of their function under pathological conditions, such as stroke and brain trauma. Using in vivo two-photon microscopy in mouse brain, we quantified mitochondrial fragmentation in a full spectrum of cortical injuries, ranging from severe to mild. Severe global ischemic injury was induced by bilateral common carotid artery occlusion, whereas severe focal stroke injury was induced by Rose Bengal photosensitization. The moderate and mild traumatic injury was inflicted by focal laser lesion and by mild photo-damage, respectively. Dendritic and mitochondrial structural changes were tracked longitudinally using transgenic mice expressing fluorescent proteins localized either in cytosol or in mitochondrial matrix. In response to severe injury, mitochondrial fragmentation developed in parallel with dendritic damage signified by dendritic beading. Reconstruction from serial section electron microscopy confirmed mitochondrial fragmentation. Unlike dendritic beading, fragmentation spread beyond the injury core in focal stroke and focal laser lesion models. In moderate and mild injury, mitochondrial fragmentation was reversible with full recovery of structural integrity after 1-2 weeks. The transient fragmentation observed in the mild photo-damage model was associated with changes in dendritic spine density without any signs of dendritic damage. Our findings indicate that alterations in neuronal mitochondria structure are very sensitive to the tissue damage and can be reversible in ischemic and traumatic injuries. During ischemic stroke or brain trauma, mitochondria can either protect neurons by supplying ATP and adsorbing excessive Ca 2+ , or kill neurons by

  1. Two-Photon Absorption Based Nanoscopic Velocimeter

    Science.gov (United States)

    Wang, Audrey; Abdalrahman, Akrm; Deng, Jianyu; Wang, Guiren

    2017-11-01

    Most velocimeters in micro/nanofluidics rely on particles as flow tracers, such as micro Particle Image Velocimetry (μPIV). However, for many microflows, such as electrokinetic and near wall flow, magnetophoresis, acoustophoresis, photophoresis and thermophoresis, particles have different velocity from their surrounding fluids. Although most molecular tracer based velocimeters can use neutral dye to measure average velocity, their temporal and spatial resolution are limited. Stimulated emission depletion (STED) based laser-induced fluorescence photobleaching anemometer (LIFPA), i.e. STED-LIFPA has achieved 70 nm spatial resolution. However, STED nanoscopy is very complicated for most users. Here we developed a two-photon absorption LIFPA (TP-LIFPA), which is relatively easier to operate. TP-LIFPA can take advantage of the two-photon microscopy to increase spatial resolution. We use a femtolaser to excite a dye. A microcapillary tube is used to test the feasibility of TP-LIFPA. TP-LIFPA can successfully measure the velocity profile in the capillary. The resolution of TP-LIFPA is estimated to be about 90 nm. The work indicates TP-LIFPA is a new promising nanoscopic velocimeter for interfacial flows, especially within 100 nm at the interfacial area between two phases in the future. The work was supported by NSF under Grant No. MRI CBET-1040227.

  2. Two Photon Exchange for Exclusive Pion Electroproduction

    Energy Technology Data Exchange (ETDEWEB)

    Afanaciev, Andrei V. [George Washington U.; Aleksejevs, Aleksandrs G. [Memorial University of Newfoundland, Newfoundland, Canada; Barkanova, Svetlana G. [Acadia University, Nova Scotia, Canada

    2013-09-01

    We perform detailed calculations of two-photon-exchange QED corrections to the cross section of pion electroproduction. The results are obtained with and without the soft-photon approximation; analytic expressions for the radiative corrections are derived. The relative importance of the two-photon correction is analyzed for the kinematics of several experiments at Jefferson Lab. A significant, over 20%, effect due to two-photon exchange is predicted for the backward angles of electron scattering at large transferred momenta.

  3. 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...

  4. Particle Tracking Facilitates Real Time Capable Motion Correction in 2D or 3D Two-Photon Imaging of Neuronal Activity

    Directory of Open Access Journals (Sweden)

    Samira Aghayee

    2017-08-01

    Full Text Available The application of 2-photon laser scanning microscopy (TPLSM techniques to measure the dynamics of cellular calcium signals in populations of neurons is an extremely powerful technique for characterizing neural activity within the central nervous system. The use of TPLSM on awake and behaving subjects promises new insights into how neural circuit elements cooperatively interact to form sensory perceptions and generate behavior. A major challenge in imaging such preparations is unavoidable animal and tissue movement, which leads to shifts in the imaging location (jitter. The presence of image motion can lead to artifacts, especially since quantification of TPLSM images involves analysis of fluctuations in fluorescence intensities for each neuron, determined from small regions of interest (ROIs. Here, we validate a new motion correction approach to compensate for motion of TPLSM images in the superficial layers of auditory cortex of awake mice. We use a nominally uniform fluorescent signal as a secondary signal to complement the dynamic signals from genetically encoded calcium indicators. We tested motion correction for single plane time lapse imaging as well as multiplane (i.e., volume time lapse imaging of cortical tissue. Our procedure of motion correction relies on locating the brightest neurons and tracking their positions over time using established techniques of particle finding and tracking. We show that our tracking based approach provides subpixel resolution without compromising speed. Unlike most established methods, our algorithm also captures deformations of the field of view and thus can compensate e.g., for rotations. Object tracking based motion correction thus offers an alternative approach for motion correction, one that is well suited for real time spike inference analysis and feedback control, and for correcting for tissue distortions.

  5. Particle Tracking Facilitates Real Time Capable Motion Correction in 2D or 3D Two-Photon Imaging of Neuronal Activity.

    Science.gov (United States)

    Aghayee, Samira; Winkowski, Daniel E; Bowen, Zachary; Marshall, Erin E; Harrington, Matt J; Kanold, Patrick O; Losert, Wolfgang

    2017-01-01

    The application of 2-photon laser scanning microscopy (TPLSM) techniques to measure the dynamics of cellular calcium signals in populations of neurons is an extremely powerful technique for characterizing neural activity within the central nervous system. The use of TPLSM on awake and behaving subjects promises new insights into how neural circuit elements cooperatively interact to form sensory perceptions and generate behavior. A major challenge in imaging such preparations is unavoidable animal and tissue movement, which leads to shifts in the imaging location (jitter). The presence of image motion can lead to artifacts, especially since quantification of TPLSM images involves analysis of fluctuations in fluorescence intensities for each neuron, determined from small regions of interest (ROIs). Here, we validate a new motion correction approach to compensate for motion of TPLSM images in the superficial layers of auditory cortex of awake mice. We use a nominally uniform fluorescent signal as a secondary signal to complement the dynamic signals from genetically encoded calcium indicators. We tested motion correction for single plane time lapse imaging as well as multiplane (i.e., volume) time lapse imaging of cortical tissue. Our procedure of motion correction relies on locating the brightest neurons and tracking their positions over time using established techniques of particle finding and tracking. We show that our tracking based approach provides subpixel resolution without compromising speed. Unlike most established methods, our algorithm also captures deformations of the field of view and thus can compensate e.g., for rotations. Object tracking based motion correction thus offers an alternative approach for motion correction, one that is well suited for real time spike inference analysis and feedback control, and for correcting for tissue distortions.

  6. An efficient two-photon fluorescent probe for human NAD(P)H:quinone oxidoreductase (hNQO1) detection and imaging in tumor cells.

    Science.gov (United States)

    Kwon, Nahyun; Cho, Myoung Ki; Park, Sang Jun; Kim, Dayoung; Nam, Sang-Jip; Cui, Lei; Kim, Hwan Myung; Yoon, Juyoung

    2017-01-03

    A new quinone propionic acid locked TP fluorophore which can be used for human NAD(P)H:quinone oxidoreductase (hNQO1) detection was developed. The probe, TPQ, which displays high selectivity and anti-interference ability, was successfully applied to endogenous hNQO1 imaging and for the identification of different cancer cells.

  7. Unravelling the Mechanism of Resonant Two-Photon Photodetachment of the Vinylidene Anion, H_2C=C^-, Using Velocity-Map Photoelectron Imaging

    Science.gov (United States)

    Gerardi, H. K.; Breen, K. J.; Gardenier, G. H.; Guasco, T. L.; Laaser, J. E.; Weddle, G. H.; Johnson, M. A.

    2009-06-01

    Vinylidene, an isomer of acetylene and the simplest unsaturated carbene, has been studied extensively due to its role as a possible intermediate in a wide range of important chemical reactions. The neutral form of vinylidene is an extremely short-lived species and therefore information about its structure and potential energy surface may be gained through the study of its stable anionic form. Using velocity-map photoelectron imaging to investigate the behavior of the anion, we observe electron loss with excitation energies lower than the established electron adiabatic detachment energy (ADE). An in-depth analysis of the velocity-map images and corresponding photoelectron spectra taken at various energies within the C-H stretching region as well as at energies around the ADE provide plausible explanations for the observed behavior. The study also reveals interesting angular distributions specific to certain vibrational modes that are not trivial to understand at this point.

  8. 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.

  9. Two-photon excited hemoglobin fluorescence

    OpenAIRE

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

    2010-01-01

    We discovered that hemoglobin emits high energy Soret fluorescence when two-photon excited by the visible femtosecond light sources. The unique spectral and temporal characteristics of hemoglobin fluorescence were measured by using a time-resolved spectroscopic detection system. The high energy Soret fluorescence of hemoglobin shows the spectral peak at 438 nm with extremely short lifetime. This discovery enables two-photon excitation fluorescence microscopy to become a potentially powerful t...

  10. 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.

  11. Spooky Phenomena in Two-Photon Processes

    Science.gov (United States)

    Li, Ming-Chiang

    2006-05-01

    A spooky phenomenon in two-photon coherent atomic absorption was discussed in 1980 [M. C. Li, Phys. Rev. A 22 (1980) 1323]. The absorption was initiated by two different laser sources. Classically, it is impossible for atoms to transit coherently in the absorption process, but quantum mechanically it is. This is one of the spooky phenomena in quantum mechanic. Around1990, there were very active experimental pursuits on a spooky phenomenon of two photons emitted from crystal parametric down conversion. The two-photon coherent atomic absorption process contained all basic ingredients as that in crystal parametric down conversion. However, the former arises from two different laser sources. The atom entangles two photons together and becomes a correlatior. The latter arises from a single laser source and two photons are entangled with each other at emission. These two spooky phenomena have been considered as disjointed. The present talk will review two spooky phenomena, and point out their similarities. The investigation on quantum spooky phenomena has led to quantum computing and quantum encryption. It is a hope that the present will stimulate the interest on bring in these two disjointed phenomena together and provide clues in advancing quantum computing and quantum encryption.

  12. 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.

  13. Signatures of plasmoemission in two photon photoemission electron microscopy

    Science.gov (United States)

    Meyer zu Heringdorf, Frank-J.; Kahl, Philip; Makris, Andreas; Sindermann, Simon; Podbiel, Daniel; Horn-von Hoegen, Michael

    2015-03-01

    The imaging of surface plasmon polariton waves in two photon photoemission microscopy has been intensely studied during the past years, with a focus on contrast mechanisms and light-plasmon interaction. The possibility of photoemission from the plasmonic fields alone has so far not been addressed in such experiments. This was justified, since the intensity of the plasmonic fields at the surface was comparatively weak and nonlinear plasmonic effects were not to be expected. Here we discuss the properties of grating couplers for creation of intense and short plasmon polariton pulses for which the emission of electrons purely from the plasmonic field cannot be neglected any more. Two examples for signatures of such nonlinear plasmoemission effects in experimental two photon photoemission microscopy images are discussed.

  14. Transition dynamics in two-photon ionisation

    Science.gov (United States)

    Vacher, Morgane; Gaillac, Romain; Maquet, Alfred; Taïeb, Richard; Caillat, Jérémie

    2017-11-01

    We review various aspects of photoemission dynamics in the case of two-photon ionisation. We first recall the definition of a transition phase specific to two-photon transitions. Numerical experiments on model atoms are used to show how the group delay associated with the transition phase is actually representative of the early dynamics of the detected photoelectron wave packets. Then we address the question of measuring these transition delays using a standard interferometric technique of experimental attosecond physics, so-called rabbit. Finally, we outline different reinterpretations of rabbit giving access to the more fundamental scattering dynamics affecting any photoemission processes.

  15. High-resolution three-dimensional imaging of a depleted CMOS sensor using an edge Transient Current Technique based on the Two Photon Absorption process (TPA-eTCT)

    CERN Document Server

    García, Marcos Fernández; Echeverría, Richard Jaramillo; Moll, Michael; Santos, Raúl Montero; Moya, David; Pinto, Rogelio Palomo; Vila, Iván

    2016-01-01

    For the first time, the deep n-well (DNW) depletion space of a High Voltage CMOS sensor has been characterized using a Transient Current Technique based on the simultaneous absorption of two photons. This novel approach has allowed to resolve the DNW implant boundaries and therefore to accurately determine the real depleted volume and the effective doping concentration of the substrate. The unprecedented spatial resolution of this new method comes from the fact that measurable free carrier generation in two photon mode only occurs in a micrometric scale voxel around the focus of the beam. Real three-dimensional spatial resolution is achieved by scanning the beam focus within the sample.

  16. High-resolution three-dimensional imaging of a depleted CMOS sensor using an edge Transient Current Technique based on the Two Photon Absorption process (TPA-eTCT)

    Energy Technology Data Exchange (ETDEWEB)

    García, Marcos Fernández; Sánchez, Javier González; Echeverría, Richard Jaramillo [Instituto de Física de Cantabria (CSIC-UC), Avda. los Castros s/n, E-39005 Santander (Spain); Moll, Michael [CERN, Organisation europénne pour la recherche nucléaire, CH-1211 Genéve 23 (Switzerland); Santos, Raúl Montero [SGIker Laser Facility, UPV/EHU, Sarriena, s/n - 48940 Leioa-Bizkaia (Spain); Moya, David [Instituto de Física de Cantabria (CSIC-UC), Avda. los Castros s/n, E-39005 Santander (Spain); Pinto, Rogelio Palomo [Departamento de Ingeniería Electrónica, Escuela Superior de Ingenieros Universidad de Sevilla (Spain); Vila, Iván [Instituto de Física de Cantabria (CSIC-UC), Avda. los Castros s/n, E-39005 Santander (Spain)

    2017-02-11

    For the first time, the deep n-well (DNW) depletion space of a High Voltage CMOS sensor has been characterized using a Transient Current Technique based on the simultaneous absorption of two photons. This novel approach has allowed to resolve the DNW implant boundaries and therefore to accurately determine the real depleted volume and the effective doping concentration of the substrate. The unprecedented spatial resolution of this new method comes from the fact that measurable free carrier generation in two photon mode only occurs in a micrometric scale voxel around the focus of the beam. Real three-dimensional spatial resolution is achieved by scanning the beam focus within the sample.

  17. Multilayer Microcapsules for FRET Analysis and Two-Photon-Activated Photodynamic Therapy.

    Science.gov (United States)

    Yang, Yang; Liu, Huiling; Han, Mingjuan; Sun, Bingbing; Li, Junbai

    2016-10-17

    Microcapsules obtained by layer-by-layer assembly provide a good platform for biological analysis owing to their component diversity, multiple binding sites, and controllable wall thickness. Herein, different assembly species were obtained from two-photon dyes and traditional photosensitizers, and further assembled into microcapsules. Fluorescence resonance energy transfer (FRET) was shown to occur between the two-photon dyes and photosensitizers. Confocal laser scanning microscopy (CLSM) with one- and two-photon lasers, fluorescence lifetime imaging microscopy (FLIM), and time-resolved fluorescence spectroscopy were used to analyze the FRET effects in the microcapsules. The FRET efficiency could easily be controlled through changing the assembly sequence. Furthermore, the capsules are phototoxic upon one- or two-photon excitation. These materials are thus expected to be applicable in two-photon-activated photodynamic therapy for deep-tissue treatment. © 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Two photon states in alkali metal clusters

    Energy Technology Data Exchange (ETDEWEB)

    Leonardi, R.; Lipparini, E. (Trento Univ. (Italy). Dipartimento di Fisica Istituto Nazionale di Fisica Nucleare, Trento (Italy))

    1991-09-01

    We study the problem of two-photon absorption in alkali metal cluster: a) by using a sum rule approach for double dipole excitation operators to have some insight on the nature of the corresponding excited states b) by using a simple random phase approximation (RPA) model to develope an harmonic model for the excited states of the system which allows an explicit prediction for the probability to excite states in the vibrational band build on the surface plasma resonance. (orig.).

  19. Two photon states in alkali metal clusters

    Science.gov (United States)

    Leonardi, R.; Lipparini, E.

    1991-09-01

    We study the problem of two-photon absorption in alkali metal clusters: a) by using a sum rule approach for double dipole excitation operators to have some insight on the nature of the corresponding excited states; b) by using a simple random phase approximation [RPA] model to develope an harmonic model for the excited states of the system which allows an explicit prediction for the probability to excite states in the vibrational band build on the surface plasma resonance.

  20. Two-Photon Fluorescence Microscopy Developed for Microgravity Fluid Physics

    Science.gov (United States)

    Fischer, David G.; Zimmerli, Gregory A.; Asipauskas, Marius

    2004-01-01

    Recent research efforts within the Microgravity Fluid Physics Branch of the NASA Glenn Research Center have necessitated the development of a microscope capable of high-resolution, three-dimensional imaging of intracellular structure and tissue morphology. Standard optical microscopy works well for thin samples, but it does not allow the imaging of thick samples because of severe degradation caused by out-of-focus object structure. Confocal microscopy, which is a laser-based scanning microscopy, provides improved three-dimensional imaging and true optical sectioning by excluding the out-of-focus light. However, in confocal microscopy, out-of-focus object structure is still illuminated by the incoming beam, which can lead to substantial photo-bleaching. In addition, confocal microscopy is plagued by limited penetration depth, signal loss due to the presence of a confocal pinhole, and the possibility of live-cell damage. Two-photon microscopy is a novel form of laser-based scanning microscopy that allows three-dimensional imaging without many of the problems inherent in confocal microscopy. Unlike one-photon microscopy, it utilizes the nonlinear absorption of two near-infrared photons. However, the efficiency of two-photon absorption is much lower than that of one-photon absorption because of the nonlinear (i.e., quadratic) electric field dependence, so an ultrafast pulsed laser source must typically be employed. On the other hand, this stringent energy density requirement effectively localizes fluorophore excitation to the focal volume. Consequently, two-photon microscopy provides optical sectioning and confocal performance without the need for a signal-limiting pinhole. In addition, there is a reduction in photo-damage because of the longer excitation wavelength, a reduction in background fluorescence, and a 4 increase in penetration depth over confocal methods because of the reduction in Rayleigh scattering.

  1. Two-Photon Fluorescence Microscope for Microgravity Research

    Science.gov (United States)

    Fischer, David G.; Zimmerli, Gregory A.; Asipauskas, Marius

    2005-01-01

    A two-photon fluorescence microscope has been developed for the study of biophysical phenomena. Two-photon microscopy is a novel form of laser-based scanning microscopy that enables three-dimensional imaging without many of the problems inherent in confocal microscopy. Unlike one-photon optical microscopy, two-photon microscopy utilizes the simultaneous nonlinear absorption of two near-infrared photons. However, the efficiency of two-photon absorption is much lower than that of one-photon absorption, so an ultra-fast pulsed laser source is typically employed. On the other hand, the critical energy threshold for two-photon absorption leads to fluorophore excitation that is intrinsically localized to the focal volume. Consequently, two-photon microscopy enables optical sectioning and confocal performance without the need for a signal-limiting pinhole. In addition, there is a reduction (relative to one-photon optical microscopy) in photon-induced damage because of the longer excitation wavelength. This reduction is especially advantageous for in vivo studies. Relative to confocal microscopy, there is also a reduction in background fluorescence, and, because of a reduction in Rayleigh scattering, there is a 4 increase of penetration depth. The prohibitive cost of a commercial two-photon fluorescence-microscope system, as well as a need for modularity, has led to the construction of a custom-built system (see Figure 1). This system includes a coherent mode-locked titanium: sapphire laser emitting 120-fs-duration pulses at a repetition rate of 80 MHz. The pulsed laser has an average output power of 800 mW and a wavelength tuning range of 700 to 980 nm, enabling the excitation of a variety of targeted fluorophores. The output from the laser is attenuated, spatially filtered, and then directed into a confocal scanning head that has been modified to provide for side entry of the laser beam. The laser output coupler has been replaced with a dichroic filter that reflects the

  2. A Two-Photon Fluorescent Probe for Lysosomal Thiols in Live Cells and Tissues

    Science.gov (United States)

    Fan, Jiangli; Han, Zhichao; Kang, Yao; Peng, Xiaojun

    2016-01-01

    Lysosome-specific fluorescent probes are exclusive to elucidate the functions of lysosomal thiols. Moreover, two-photon microscopy offers advantages of less phototoxicity, better three dimensional spatial localization, deeper penetration depth and lower self-absorption. However, such fluorescent probes for thiols are still rare. In this work, an efficient two-photon fluorophore 1,8-naphthalimide-based probe conjugating a 2,4-dinitrobenzenesulfonyl chloride and morpholine was designed and synthesized, which exhibited high selectivity and sensitivity towards lysosomal thiols by turn-on fluorescence method quantitatively and was successfully applied to the imaging of thiols in live cells and tissues by two-photon microscopy.

  3. Two-Photon Small Molecule Enzymatic Probes.

    Science.gov (United States)

    Qian, Linghui; Li, Lin; Yao, Shao Q

    2016-04-19

    Enzymes are essential for life, especially in the development of disease and on drug effects, but as we cannot yet directly observe the inside interactions and only partially observe biochemical outcomes, tools "translating" these processes into readable information are essential for better understanding of enzymes as well as for developing effective tools to fight against diseases. Therefore, sensitive small molecule probes suitable for direct in vivo monitoring of enzyme activities are ultimately desirable. For fulfilling this desire, two-photon small molecule enzymatic probes (TSMEPs) producing amplified fluorescent signals based on enzymatic conversion with better photophysical properties and deeper penetration in intact tissues and whole animals have been developed and demonstrated to be powerful in addressing the issues described above. Nonetheless, currently available TSMEPs only cover a small portion of enzymes despite the distinct advantages of two-photon fluorescence microscopy. In this Account, we would like to share design principles for TSMEPs as potential indicators of certain pathology-related biomarkers together with their applications in disease models to inspire more elegant work to be done in this area. Highlights will be addressed on how to equip two-photon fluorescent probes with features amenable for direct assessment of enzyme activities in complex pathological environments. We give three recent examples from our laboratory and collaborations in which TSMEPs are applied to visualize the distribution and activity of enzymes at cellular and organism levels. The first example shows that we could distinguish endogenous phosphatase activity in different organelles; the second illustrates that TSMEP is suitable for specific and sensitive detection of a potential Parkinson's disease marker (monoamine oxidase B) in a variety of biological systems from cells to patient samples, and the third identifies that TSMEPs can be applied to other enzyme

  4. 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...... 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....

  5. Platinum Acetylide Two-Photon Chromophores (Preprint)

    Science.gov (United States)

    2007-04-01

    L and 2-L) or 370 nm (1 and 2) with a matched optical density of 0.1. Fluorescence quantum yield for 3-L was measured relative to Coumarin 30 in...short-wavelength region, Aex= 600 - 700 run, a strong isolated 2PA peak shows up in all cases. For quantitative comparison of 2PA strength of our... isolated peaks at 300 - 350 nm in IPA). This 2PA peak can be tentatively assigned to a one-photon forbidden, but two-photon allowed gerade-gerade

  6. Compact two-photon fluorescence microscope based on a single-mode fiber coupler.

    Science.gov (United States)

    Bird, Damian; Gu, Min

    2002-06-15

    We present a two-photon fluorescence microscope based on a three-port single-mode optical fiber coupler. It is found that the coupler behaves as a low-pass filter that can deliver an ultrashort-pulsed laser beam of as much as 150 mW of power in the wavelength range from 770 to 870 nm as well as collect a two-photon fluorescence signal in the visible range. As a result of using the fiber coupler, the new two-photon imaging system exhibts a number of advantages, including a compact arrangement, freedom from vibration from lasers and electronic devices, self-alignment, reduction of multiple scattering, and an enhanced optical sectioning effect. The effectiveness of the new instrument is demonstrated with a set of three-dimensional images of biological samples. This instrument may make two-photon fluorescence endoscopy possible for in vivo medical applications.

  7. Two-color two-photon fluorescence laser scanning microscopy.

    Science.gov (United States)

    Quentmeier, S; Denicke, S; Gericke, K-H

    2009-11-01

    We present the first realization of a Two-Color Two-Photon Laser-Scanning Microscope (2c2pLSM) and UV fluorescence images of cells acquired with this technique. Fluorescence is induced by two-color two-photon absorption using the fundamental and the second harmonic of a Ti:Sa femtosecond laser. Simultaneous absorption of an 800 nm photon and a 400 nm photon energetically corresponds to one-photon absorption at 266 nm. This technique for Laser-Scanning Microscopy extends the excitation wavelength range of a Ti:Sa powered fluorescence microscope to the UV. In addition to the known advantages of multi-photon microscopy like intrinsic 3D resolution, reduced photo damage and high penetration depth 2c2pLSM offers the possibility of using standard high numeric aperture objectives for UV fluorescence imaging. The effective excitation wavelength of 266 nm corresponds especially well to the excitation spectrum of tryptophan. Hence, it is an ideal tool for label free fluorescence studies and imaging of intrinsic protein fluorescence which originates mainly from tryptophan. Thus a very sensitive natural lifetime probe can be used for monitoring protein reactions or changes in conformation. First measurements of living MIN-6 cells reveal differences between the UV fluorescence lifetimes of the nucleus and cytoplasm. The significance of this method was further demonstrated by monitoring the binding of biotin to avidin.

  8. 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

  9. Quantum fingerprinting using two-photon interference.

    Science.gov (United States)

    Jachura, Michał; Lipka, Michał; Jarzyna, Marcin; Banaszek, Konrad

    2017-10-30

    We present a quantum fingerprinting protocol relying on two-photon interference which does not require a shared phase reference between the parties preparing optical signals carrying data fingerprints. We show that the scaling of the protocol, in terms of transmittable classical information, is analogous to the recently proposed and demonstrated scheme based on coherent pulses and first-order interference, offering comparable advantage over classical fingerprinting protocols without access to shared prior randomness. We analyze the protocol taking into account non-Poissonian photon statistics of optical signals and a variety of imperfections, such as transmission losses, dark counts, and residual distinguishability. The impact of these effects on the protocol performance is quantified with the help of Chernoff information.

  10. 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.

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

    Science.gov (United States)

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

    2017-03-01

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

  12. Enhancement of two-photon photoluminescence and SERS for low-coverage gold films

    DEFF Research Database (Denmark)

    Novikov, Sergey M.; Beermann, Jonas; Frydendahl, Christian

    2016-01-01

    Electromagnetic field enhancement (FE) effects occurring in thin gold films 3-12-nm are investigated with two-photon photoluminescence (TPL) and Raman scanning optical microscopies. The samples are characterized using scanning electron microscopy images and linear optical spectroscopy. TPL images...

  13. Development of a two photon microscope for tracking Drosophila larvae

    Science.gov (United States)

    Karagyozov, Doycho; Mihovilovic Skanata, Mirna; Gershow, Marc

    Current in vivo methods for measuring neural activity in Drosophila larva require immobilization of the animal. Although we can record neural signals while stimulating the sensory organs, we cannot read the behavioral output because we have prevented the animal from moving. Many research questions cannot be answered without observation of neural activity in behaving (freely-moving) animals. We incorporated a Tunable Acoustic Gradient (TAG) lens into a two-photon microscope to achieve a 70kHz axial scan rate, enabling volumetric imaging at tens of hertz. We then implemented a tracking algorithm based on a Kalman filter to maintain the neurons of interest in the field of view and in focus during the rapid three dimensional motion of a free larva. Preliminary results show successful tracking of a neuron moving at speeds reaching 500 μm/s. NIH Grant 1DP2EB022359 and NSF Grant PHY-1455015.

  14. Rapid three dimensional two photon neural population scanning.

    Science.gov (United States)

    Schuck, Renaud; Quicke, Peter; Copeland, Caroline; Garasto, Stefania; Annecchino, Luca A; Hwang, June Kyu; Schultz, Simon R

    2015-08-01

    Recording the activity of neural populations at high sampling rates is a fundamental requirement for understanding computation in neural circuits. Two photon microscopy provides one promising approach towards this. However, neural circuits are three dimensional, and functional imaging in two dimensions fails to capture the 3D nature of neural dynamics. Electrically tunable lenses (ETLs) provide a simple and cheap method to extend laser scanning microscopy into the relatively unexploited third dimension. We have therefore incorporated them into our Adaptive Spiral Scanning (SSA) algorithm, which calculates kinematically efficient scanning strategies using radially modulated spiral paths. We characterised the response of the ETL, incorporated its dynamics using MATLAB models of the SSA algorithm and tested the models on populations of Izhikevich neurons of varying size and density. From this, we show that our algorithms can theoretically at least achieve sampling rates of 36.2Hz compared to 21.6Hz previously reported for 3D scanning techniques.

  15. High visibility two-photon interference with classical light.

    Science.gov (United States)

    Hong, Peilong; Xu, Lei; Zhai, Zhaohui; Zhang, Guoquan

    2013-06-17

    Two-photon interference with independent classical sources, in which superposition of two indistinguishable two-photon paths plays a key role, is of limited visibility with a maximum value of 50%. By using a random-phase grating to modulate the wavefront of a coherent light, we introduce superposition of multiple indistinguishable two-photon paths, which enhances the two-photon interference effect with a signature of visibility exceeding 50%. The result shows the importance of phase control in the control of high-order coherence of classical light.

  16. Two-photon fluorescence microscope with a hollow-core photonic crystal fiber.

    Science.gov (United States)

    Tai, Shih-Peng; Chan, Ming-Che; Tsai, Tsung-Han; Guol, Shi-Hao; Chen, Li-Jin; Sun, Chi-Kuang

    2004-12-13

    Self-phase-modulation and group velocity dispersion of near IR femtosecond pulses in fibers restrict their use in two-photon fluorescence microscopy (TPFM). Here we demonstrate a hollow-core photonic crystal fiber based two-photon fluorescence microscope with low nonlinearity and dispersion effects. We use this fiber-based TPFM system to take two-photon fluorescence (chlorophyll) images of mesophyll tissue in the leaf of Rhaphidophora aurea. With less than 2mW average power exposure on the leaf at 755nm, the near zero-dispersion wavelength, chloroplasts distribution inside the mesophyll cells can be identified with a sub-micron spatial resolution. The acquired image quality is comparable to that acquired by the conventional fiber-free TPFM system, due to the negligible temporal pulse broadening effect.

  17. Two-photon exchange effect on deuteron electromagnetic form factors

    OpenAIRE

    Dong, Yu Bing; Chen, D. Y.

    2009-01-01

    Corrections of two-photon exchange to proton and neutron electromagnetic form factors are employed to study the effect of two-photon exchange on the deuteron electromagnetic form factors. Numerical results of the effect are given. It is suggested to test the effect in the measurement of $P_z$ in a small angle limit.

  18. Two-photon coherent states of the radiation field

    Science.gov (United States)

    Yuen, H. P.

    1976-01-01

    The concept of a two-photon coherent state is introduced for applications in quantum optics. It is a simple generalization of the well-known minimum-uncertainty wave packets. The detailed properties of two-photon coherent states are developed and distinguished from ordinary coherent states. These two-photon coherent states are mathematically generated from coherent states through unitary operators associated with quadratic Hamiltonians. Physically they are the radiation states of ideal two-photon lasers operating far above threshold, according to the self-consistent-field approximation. The mean-square quantum noise behavior of these states, which is basically the same as those of minimum-uncertainty states, leads to applications not obtainable from coherent states or one-photon lasers. The essential behavior of two-photon coherent states is unchanged by small losses in the system. The counting rates or distributions these states generate in photocount experiments also reveal their difference from coherent states.

  19. Twofold spatial resolution enhancement by two-photon exposure of photographic film

    Science.gov (United States)

    Korobkin, Dmitriy V.; Yablonovitch, Eli

    2002-07-01

    Two-photon absorption of photosensitive media can produce interference fringes with double spatial frequency. This requires the employment of multiple-frequency beams, which interfere with one another to produce a stationary image with double spatial resolution. The required beams were produced by frequency filtering of broadband radiation from a cw mode-locked femtosecond Ti:sapphire laser ((lambda) equals 790 nm) in a dispersion-free pulse shaper. Then the two multifrequency rays converged from opposite edges of a lens, focusing on Kodak commercial film. The laser intensity was high enough to produce a two-photon exposure. The doubling of the spatial frequency of the interference pattern has been observed, but the contrast ratio of the pattern was limited by competition from the more usual one-photon absorption. Laser pulse parameters for a single-pulse two-photon exposure have been estimated.

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

    DEFF Research Database (Denmark)

    Biagioni, P.; Celebrano, M.; Savoini, M.

    2009-01-01

    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...

  1. Feasibility of using gold nanorods for optical contrast in two photon microscopy of oral carcinogenesis

    Science.gov (United States)

    Motamedi, Saam; Shilagard, Tuya; Koong, Luke; Vargas, Gracie

    2010-02-01

    Gold nanorods (GNRs) combined with two-photon microscopy were explored for potential application in imaging of oral carcinogenesis. GNRs have been shown to be effective contrast agents for two photon luminescence in that excitation laser powers required for imaging are low compared to traditional fluorophores. Imaging of cells, ex vivo tissues, and in vivo oral mucosa labeled with GNRs was performed to evaluate potential advantages of these agents in molecular imaging of epithelial carcinogenesis. Powers required to elicit a two-photon luminescence signal from GNRs were determined for cells as well as normal and malignant transformed lesions, 24 hours following injection of GNRs in a hamster model for oral cancer. The strength of the detected emission as the function of the average incident laser power was measured in tissues with and without GNRs to compare the sensitivity of GNRs against tissue autofluorescence. Finally, in vivo imaging was performed immediately following GNR injection to establish the ability to image microvasculature at low incident powers. The pilot study demonstrated uptake of GNRs by cells and in tissues yielding bright fluorescence signals using significantly lower incident powers than those needed to excite tissue autofluorescence. The in vivo imaging aspect of the study demonstrated the localization of GNRs within the microvasculature of the oral cancer model. These preliminary studies demonstrated the ability of GNRs to function as photostable, high contrast imaging agents and suggest that GNRs and multi-photon imaging have great potential for applications in the field of molecular imaging and early detection of cancer.

  2. Two-photon quantum Rabi model with superconducting circuits

    Science.gov (United States)

    Felicetti, S.; Rossatto, D. Z.; Rico, E.; Solano, E.; Forn-Díaz, P.

    2018-01-01

    We propose a superconducting circuit to implement a two-photon quantum Rabi model in a solid-state device, where a qubit and a resonator are coupled by a two-photon interaction. We analyze the input-output relations for this circuit in the strong-coupling regime and find that fundamental quantum-optical phenomena are qualitatively modified. For instance, two-photon interactions are shown to yield a single- or two-photon blockade when a pumping field is either applied to the cavity mode or to the qubit, respectively. In addition, we derive an effective Hamiltonian for perturbative ultrastrong two-photon couplings in the dispersive regime, where two-photon interactions introduce a qubit-state-dependent Kerr term. Finally, we analyze the spectral collapse of the multiqubit two-photon quantum Rabi model and find a scaling of the critical coupling with the number of qubits. Using realistic parameters with the circuit proposed, three qubits are sufficient to reach the collapse point.

  3. 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.

  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. Phase Measurement of Resonant Two-Photon Ionization in Helium

    CERN Document Server

    Swoboda, M; Klünder, K; Dahlström, J M; Miranda, M; Buth, C; Schafer, K J; Mauritsson, J; L'Huillier, A; Gisselbrecht, M

    2010-01-01

    We study resonant two-color two-photon ionization of Helium via the 1s3p 1P1 state. The first color is the 15th harmonic of a tunable titanium sapphire laser, while the second color is the fundamental laser radiation. Our method uses phase-locked high-order harmonics to determine the {\\it phase} of the two-photon process by interferometry. The measurement of the two-photon ionization phase variation as a function of detuning from the resonance and intensity of the dressing field allows us to determine the intensity dependence of the transition energy.

  6. Femtosecond photoacoustics: integrated two-photon fluorescence and photoacoustic microscopy

    Science.gov (United States)

    van Raaij, Martijn E.; Lee, Mike; Chérin, Emmanuel; Stefanovic, Bojana; Foster, F. Stuart

    2010-02-01

    Conventional photoacoustic imaging systems excite a photoacoustic wave by illuminating an area on the order of square centimeters with millijoule laser pulses. Spatial resolution is then determined by the ultrasound transducer and is typically on the order of 100 μm. We report on a system that focuses femtosecond, nanojoule pulses to a spot with a diameter of ~ 1 μm to perform laser-scanning photoacoustics with micrometer resolution. Near-infrared femtosecond laser pulses with a pulse energy of 2.4 nanojoules excite a train of photoacoustic waves at the repetition rate of the pulsed laser (80 MHz). These photoacoustic waves are detected by an unfocused single-element ultrasound transducer tuned to 80 MHz. A radiofrequency lock-in amplifier recovers the amplitude of the frequency component of the photoacoustic signal at the pulse repetition frequency. This amplitude is an indicator of the absorption coefficient of the sample at the laser focus and at the laser wavelength. Initial experiments using a graphite rod as absorber reproducibly yield signals in the 0.2 - 2 microvolt range with a signal-to-noise ratio of 18 dB, recovered from 10 mV of broadband noise. The photoacoustic imaging system is integrated in a commercial laser-scanning two-photon fluorescence microscope, enabling simultaneous three-dimensional fluorescence- and photoacoustic imaging. One major application will be to image both morphology and oxygen saturation of microvasculature in the cerebral cortex of anesthetized rodents in vivo in the context of tumor angiogenesis. In this paper we describe the physics of femtosecond photoacoustics and demonstrate initial results.

  7. 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.

  8. Two-Photon Fluorescence Microscopy for Biomedical Research

    Science.gov (United States)

    Fischer, David; Zimmerli, Greg; Asipauskas, Marius

    2007-01-01

    This viewgraph presentation gives an overview of two-photon microscopy as it applies to biomedical research. The topics include: 1) Overview; 2) Background; 3) Principles of Operation; 4) Advantages Over Confocal; 5) Modes of Operation; and 6) Applications.

  9. Two-photon physics at RHIC: Separating signals from backgrounds

    Energy Technology Data Exchange (ETDEWEB)

    Nystrand, J.; Klein, S.

    1997-11-01

    This presentation will show the feasibility of studying two-photon interactions in the STAR experiment at RHIC. Signals, detection efficiencies, backgrounds, triggering and analysis techniques will be discussed.

  10. 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.

    2015-01-01

    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. PMID:25836193

  11. 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.

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

    DEFF Research Database (Denmark)

    Nielsen, Christian Benedikt

    amount of charge-transfer is beneficial for high two-photon absorption cross sections but iscounter-productive for singlet oxygen generation. The design principles obtained from the studies in lipophilic solvents were applied to synthesize water-soluble twophoton singlet oxygen sensitizers......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...

  13. 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.

  14. Two Photon Polymerization of Microneedles for Transdermal Drug Delivery

    Science.gov (United States)

    Gittard, Shaun D.; Ovsianikov, Aleksandr; Chichkov, Boris N.; Doraiswamy, Anand; Narayan, Roger J.

    2010-01-01

    Importance of the field Microneedles are small-scale devices that are finding use for transdermal delivery of protein-based pharmacologic agents and nucleic acid-based pharmacologic agents; however, microneedles prepared using conventional microelectronics-based technologies have several shortcomings, which have limited translation of these devices into widespread clinical use. Areas covered in this review Two photon polymerization is a laser-based rapid prototyping technique that has been recently used for direct fabrication of hollow microneedles with a wide variety of geometries. In addition, an indirect rapid prototyping method that involves two photon polymerization and polydimethyl siloxane micromolding has been used for fabrication of solid microneedles with exceptional mechanical properties. What the reader will gain In this review, the use of two photon polymerization for fabricating in-plane and out-of-plane hollow microneedle arrays is described. The use of two photon polymerization-micromolding for fabrication of solid microneedles is also reviewed. In addition, fabrication of microneedles with antimicrobial properties is discussed; antimicrobial microneedles may reduce the risk of infection associated with formation of channels through the stratum corneum. Take home message It is anticipated that the use of two photon polymerization as well as two photon polymerization-micromolding for fabrication of microneedles and other microstructured drug delivery devices will increase over the coming years. PMID:20205601

  15. Two-photon polymerization of microneedles for transdermal drug delivery.

    Science.gov (United States)

    Gittard, Shaun D; Ovsianikov, Aleksandr; Chichkov, Boris N; Doraiswamy, Anand; Narayan, Roger J

    2010-04-01

    Microneedles are small-scale devices that are finding use for transdermal delivery of protein-based pharmacologic agents and nucleic acid-based pharmacologic agents; however, microneedles prepared using conventional microelectronics-based technologies have several shortcomings, which have limited translation of these devices into widespread clinical use. Two-photon polymerization is a laser-based rapid prototyping technique that has been used recently for direct fabrication of hollow microneedles with a wide variety of geometries. In addition, an indirect rapid prototyping method that involves two-photon polymerization and polydimethyl siloxane micromolding has been used for fabrication of solid microneedles with exceptional mechanical properties. In this review, the use of two-photon polymerization for fabricating in-plane and out-of-plane hollow microneedle arrays is described. The use of two-photon polymerization-micromolding for fabrication of solid microneedles is also reviewed. In addition, fabrication of microneedles with antimicrobial properties is discussed; antimicrobial microneedles may reduce the risk of infection associated with the formation of channels through the stratum corneum. It is anticipated that the use of two-photon polymerization as well as two-photon polymerization-micromolding for fabrication of microneedles and other microstructured drug delivery devices will increase over the coming years.

  16. 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.

  17. 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.

  18. Organelle-specific detection of phosphatase activities with two-photon fluorogenic probes in cells and tissues.

    Science.gov (United States)

    Li, Lin; Ge, Jingyan; Wu, Hao; Xu, Qing-Hua; Yao, Shao Q

    2012-07-25

    Two-photon fluorescence microscopy (TPFM) provides key advantages over conventional fluorescence imaging techniques, namely, increased penetration depth, lower tissue autofluorescence and self-absorption, and reduced photodamage and photobleaching and therefore is particularly useful for imaging deep tissues and animals. Enzyme-detecting, small molecule probes provide powerful alternatives over conventional fluorescent protein (FP)-based methods in bioimaging, primarily due to their favorable photophysical properties, cell permeability, and chemical tractability. In this article, we report the first fluorogenic, small molecule reporter system (Y2/Y1) capable of imaging endogenous phosphatase activities in both live mammalian cells and Drosophila brains. The one- and two-photon excited photophysical properties of the system were thoroughly investigated, thus confirming the system was indeed a suitable Turn-ON fluorescence pair for TPFM. To our knowledge, this is the first enzyme reporting two-photon fluorescence bioimaging system which was designed exclusively from a centrosymmetric dye possessing desirable two-photon properties. By conjugation of our reporter system to different cell-penetrating peptides (CPPs), we were able to achieve organelle- and tumor cell-specific imaging of phosphatase activities with good spatial and temporal resolution. The diffusion problem typically associated with most small molecule imaging probes was effectively abrogated. We further demonstrated this novel two-photon system could be used for imaging endogenous phosphatase activities in Drosophila brains with a detection depth of >100 μm.

  19. 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.

  20. 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.

  1. Selective two-photon collagen crosslinking in situ measured by Brillouin microscopy (Conference Presentation)

    Science.gov (United States)

    Kwok, Sheldon J. J.; Kuznetsov, Ivan A.; Kim, Moonseok; Choi, Myunghwan; Scarcelli, Giuliano; Yun, Seok-Hyun

    2017-02-01

    Two-photon polymerization and crosslinking are commonly used methods for microfabrication of three-dimensional structures with applications spanning from photonic microdevices, drug delivery systems, to cellular scaffolds. However, the use of two-photon processes for precise, internal modification of biological tissues has not yet been reported. One of the major challenges has been a lack of appropriate tools to monitor and characterize crosslinked regions nondestructively. Here, we demonstrate spatially selective two-photon collagen crosslinking (2P-CXL) in intact tissue for the first time. Using riboflavin photosensitizer and femtosecond laser irradiation, we crosslinked a small volume of tissue within animal corneas. Collagen fiber orientations and photobleaching were characterized by second harmonic generation and two-photon fluorescence imaging, respectively. Using confocal Brillouin microscopy, we measured local changes in longitudinal mechanical moduli and visualized the cross-linked pattern without perturbing surrounding non-irradiated regions. 2P-CXL-induced tissue stiffening was comparable to that achieved with conventional one-photon CXL. Our results demonstrate the ability to selectively stiffen biological tissue in situ at high spatial resolution, with broad implications in ophthalmology, laser surgery, and tissue engineering.

  2. Two-Photon Coherent Atomic Absorption of Multiple Laser Beams

    Science.gov (United States)

    Li, Ming-Chiang

    2006-05-01

    Physical processes on two-photon coherent atomic absorption of multiple laser beams were discussed about thirty years ago [M. C. Li, Bull. Am. Phys. Soc. 20, 654 (1975)]. These processes can be divided into two distinct groups. In the first group, laser beams are from a single source, and in the second group laser beams are from two different sources [M. C. Li, Phys. Rev. A 22 (1980) 1323]. Several experiments in the first group were carried out and have led to the 2005 Nobel Prize in physics. The second group is more interesting. Beside atoms are in random motion, two photons are from different sources. Classically, it is impossible for atoms to transit coherently in the absorption process, but quantum mechanically, such a transition is possible and that is one of the spooky phenomena in quantum mechanic. To assure the coherent transition, each photon as absorbed by the atom must have two possible paths of choices. If one photon has the choice and other one is not, then the atomic transitions cannot be coherent. Around1990, there were very active experimental pursuits on such a spooky phenomenon of two photons emitted from crystal parametric down conversion. The present talk will review various spooky phenomena associated with two-photon coherent atomic absorption. Hope that the talk will stimulate the interest on the long neglected experimental front on two-photon coherent atomic absorption from two different laser sources.

  3. Simultaneous imaging of neural activity in three dimensions

    Directory of Open Access Journals (Sweden)

    Sean eQuirin

    2014-04-01

    Full Text Available We introduce a scanless optical method to image neuronal activity in three dimensions simultaneously. Using a spatial light modulator and a custom-designed phase mask, we illuminate and collect light simultaneously from different focal planes and perform calcium imaging of neuronal activity in vitro and in vivo. This method, combining structured illumination with volume projection imaging, could be used as a technological platform for brain activity mapping.

  4. Two-photon luminescence microscopy of field enhancement at gold nanoparticles

    DEFF Research Database (Denmark)

    Beermann, Jonas; Bozhevolnyi, Sergey I.

    2005-01-01

    Using a reflection scanning optical microscope detecting two-photon luminescence (TPL) we have imaged square gold bumps positioned in a periodic array either on a smooth gold film or directly on a glass substrate. The second-harmonic (SH) and TPL response from these structures show both polarizat......Using a reflection scanning optical microscope detecting two-photon luminescence (TPL) we have imaged square gold bumps positioned in a periodic array either on a smooth gold film or directly on a glass substrate. The second-harmonic (SH) and TPL response from these structures show both...... polarization and wavelength dependence. The gold bumps on gold film showed extremely high sensitivity to the incident field, with the strongest TPL response from the gold bumps being enhanced nearly 103 times compared to the TPL response from the smooth gold surface. For gold bumps directly on glass...... the similar calculated enhancement was estimated to be ~ 10^2...

  5. 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

    2016-01-01

    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. PMID:26864567

  6. Mid-infrared two photon absorption sensitivity of commercial detectors

    Science.gov (United States)

    Boiko, D. L.; Antonov, A. V.; Kuritsyn, D. I.; Yablonskiy, A. N.; Sergeev, S. M.; Orlova, E. E.; Vaks, V. V.

    2017-10-01

    We report on broad-band two-photon absorption (TPA) in several commercially available MIR inter-band bulk semiconductor photodetectors with the spectral cutoff in the range of 4.5-6 μm. The highest TPA responsivity of 2 × 10-5 A.mm2/W2 is measured for a nitrogen-cooled InSb photovoltaic detector. Its performance as a two-photon detector is validated by measuring the second-order interferometric autocorrelation function of a multimode quantum cascade laser emitting at the wavelength of 8 μm.

  7. Two photon couplings of the lightest isoscalars from BELLE data

    Directory of Open Access Journals (Sweden)

    Ling-Yun Dai

    2014-09-01

    Full Text Available Amplitude Analysis of two photon production of ππ and K¯K, using S-matrix constraints and fitting all available data, including the latest precision results from Belle, yields a single partial wave solution up to 1.4 GeV. The two photon couplings of the σ/f0(500, f0(980 and f2(1270 are determined from the residues of the resonance poles. These amplitudes are a key input into the newly developed dispersive approach to calculating hadronic light-by-light scattering for (g−2 of the muon.

  8. A new technique for assessing hybrid layer interfacial micromorphology and integrity: two-photon laser microscopy.

    Science.gov (United States)

    D'Alpino, Paulo H P; Pereira, José C; Svizero, Nádia R; Rueggeberg, Frederick A; Carvalho, Ricardo M; Pashley, David H

    2006-10-01

    This study describes a two-photon laser fluorescence microscopy technique developed to evaluate the interfacial micromorphology of the hybrid layer in bonded restorations. Micropermeability of the hybrid layer was characterized by means of simultaneously contrasting a dye-containing adhesive with a differently colored dye placed into the pulp chamber and allowed to diffuse toward the different-colored hybrid layer. A fluorescent red dye (rhodamine B) was incorporated into a commercial dentin bonding agent. Class I preparations (margins in enamel) were made on extracted human third molars. The teeth were restored using conventional methods: bonding agent, composite, finishing, and polishing. An aqueous solution of a yellow/green dye (fluorescein) was then placed into the pulp chamber for 3 h, allowing time to diffuse toward the different-colored bonded interface. The teeth were then embedded, sectioned, and microscopically analyzed using two-photon laser microscopy at 40X magnification. Subsurface fluorescent imaging using this technique enabled interfacial micromorphology to be characterized at submicrometer resolution and provided high-contrast images. The quality of surrounding structures and potential presence of gaps were also precisely assessed. Two-photon laser microscopy provided high quality, high-resolution images of the bonded interface and surrounding areas, allowing accurate qualitative and quantitative analysis of the structure and integrity of the hybrid layer.

  9. 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.

  10. Two-Photon Interactions at Belle and BaBar

    CERN Document Server

    Eidelman, Simon

    2010-01-01

    Results on two-photon physics obtained in experiments at the B factories are discussed. BaBar used single-tag collisions to measure the transition form factor of the 0 meson. Belle studied no-tag collisions to measure cross sections of exclusive production of two baryons and two mesons. Experimental results are confronted with QCD predictions.

  11. 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...

  12. 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.

  13. New two-photon based nanoscopic modalities and optogenetics

    DEFF Research Database (Denmark)

    Glückstad, Jesper

    The science fiction inspired shrinking of macro-scale robotic manipulation and handling down to the micro- and nanoscale regime open new doors for exploiting the forces and torques of light for micro- and nanobiologic probing, actuation and control [1-3]. A generic approach for optimizing light......-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...... of two-photon polymerised microstructures equipped with features easily entering the submicron-regime. Aided by European collaborators who fabricated test structures with built-in waveguides for us, we were able to put the idea of optically steerable freestanding waveguides – coined: wave-guided optical...

  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-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).

  16. 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.

  17. Twist-three effects in two-photon processes

    Science.gov (United States)

    Belitsky, A. V.; Müller, D.

    2000-11-01

    We give a general treatment of twist-three effects in two-photon reactions. We address the issue of the gauge invariance of the Compton amplitude in generalized Bjorken kinematics and relations of twist-three `transverse' skewed parton distributions to twist-two ones and interaction dependent three-particle correlation functions. Finally, we discuss leading order evolution of twist-three functions and their impact on the deeply virtual Compton scattering.

  18. Two photon couplings of scalar and tensor mesons

    Science.gov (United States)

    Feindt, Michael; Harjes, Jens

    1991-06-01

    Experimental data on exclusive two photon reactions are investigated with respect to formation of tensor and scalar mesons. Theoretical and experimental status and progress is reviewed. Furthermore, new CELLO results on γγ → π-π- and γγ → ϱ0ϱ0 are presented. Clear evidence for a large scalar contribution is found in both reactions. The implications of these new results are discussed.

  19. 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.

  20. Spooky Phenomena in Two-Photon Coherent Atomic Absorption

    Science.gov (United States)

    Li, Ming-Chiang

    2006-03-01

    Physical processes on two-photon coherent atomic absorption of multiple laser beams were discussed more than twenty five years ago. These processes can be divided into two distinct groups. In the first group, laser beams are from a single source^1,2, and in the second group laser beams are from two different sources^3. Several experiments in the first group were carried out and have led to the 2005 Nobel Prize in physics. The second group is more interesting. Atoms are in random motion and two photons are from different sources. Classically, it is impossible for atoms to transit coherently in the absorption process, but quantum mechanically, such a transition is possible and that is one of the spooky phenomena in quantum mechanic. To assure the coherent transition, each photon as absorbed by the atom must have two possible paths of choices. If one photon has the choice and other one is not, then the atomic transitions cannot be coherent. The present talk will review various spooky phenomena associated with two-photon coherent atomic absorption, and will clarify some theoretical misunderstandings regarding these interesting transitions. Reference: *M. C. Li, Nuovo Cimento 39B (1977) 165. *M. C. Li, Phys. Rev. A 16 (1977) 2480. *M. C. Li, Phys. Rev. A 22 (1980) 1323.

  1. 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.

  2. Scanning two-photon microscopy with upconverting lanthanide nanoparticles via Richardson-Lucy deconvolution

    Science.gov (United States)

    Gainer, Christian F.; Utzinger, Urs

    2012-01-01

    Abstract. The use of upconverting lanthanide nanoparticles in fast-scanning microscopy is hindered by a long luminescence decay time, which greatly blurs images acquired in a nondescanned mode. We demonstrate herein an image processing method based on Richardson-Lucy deconvolution that mitigates the detrimental effects of their luminescence lifetime. This technique generates images with lateral resolution on par with the system’s performance, ∼1.2  μm, while maintaining an axial resolution of 5 μm or better at a scan rate comparable with traditional two-photon microscopy. Remarkably, this can be accomplished with near infrared excitation power densities of 850  W/cm2, several orders of magnitude below those used in two-photon imaging with molecular fluorophores. By way of illustration, we introduce the use of lipids to coat and functionalize these nanoparticles, rendering them water dispersible and readily conjugated to biologically relevant ligands, in this case epidermal growth factor receptor antibody. This deconvolution technique combined with the functionalized nanoparticles will enable three-dimensional functional tissue imaging at exceptionally low excitation power densities. PMID:22894486

  3. Two-photon time-lapse microscopy of BODIPY-cholesterol reveals anomalous sterol diffusion in chinese hamster ovary cells

    DEFF Research Database (Denmark)

    Lund, F. W.; Lomholt, M. A.; Solanko, L. M.

    2012-01-01

    probe has utility for prolonged live-cell imaging of sterol transport. Results: We found that BChol is very photostable under two-photon (2P)-excitation allowing the acquisition of several hundred frames without significant photobleaching. Therefore, long-term tracking and diffusion measurements...

  4. Two-photon excited fluorescence microscopy application for ex vivo investigation of ocular fundus samples

    Science.gov (United States)

    Peters, Sven; Hammer, Martin; Schweitzer, Dietrich

    2011-07-01

    Two-photon excited fluorescence (TPEF) imaging of ocular tissue has recently become a promising tool in ophthalmology for diagnostic and research purposes. The feasibility and the advantages of TPEF imaging, namely deeper tissue penetration and improved high-resolution imaging of microstructures, have been demonstrated lately using human ocular samples. The autofluorescence properties of endogenous fluorophores in ocular fundus tissue are well known from spectrophotometric analysis. But fluorophores, especially when it comes to fluorescence lifetime, typically display a dependence of their fluorescence properties on local environmental parameters. Hence, a more detailed investigation of ocular fundus autofluorescence ideally in vivo is of utmost interest. The aim of this study is to determine space-resolved the stationary and time-resolved fluorescence properties of endogenous fluorophores in ex vivo porcine ocular fundus samples by means of two-photon excited fluorescence spectrum and lifetime imaging microscopy (FSIM/FLIM). By our first results, we characterized the autofluorescence of individual anatomical structures of porcine retina samples excited at 760 nm. The fluorescence properties of almost all investigated retinal layers are relatively homogenous. But as previously unknown, ganglion cell bodies show a significantly shorter fluorescence lifetime compared to the adjacent mueller cells. Since all retinal layers exhibit bi-exponential autofluorescence decays, we were able to achieve a more precise characterization of fluorescence properties of endogenous fluorophores compared to a present in vivo FLIM approach by confocal scanning laser ophthalmoscope (cSLO).

  5. A highly specific ratiometric two-photon fluorescent probe to detect dipeptidyl peptidase IV in plasma and living systems.

    Science.gov (United States)

    Zou, Li-Wei; Wang, Ping; Qian, Xing-Kai; Feng, Lei; Yu, Yang; Wang, Dan-Dan; Jin, Qiang; Hou, Jie; Liu, Zhi-Hong; Ge, Guang-Bo; Yang, Ling

    2017-04-15

    In this study, a highly specific ratiometric two-photon fluorescent probe GP-BAN was developed and well-characterized to monitor dipeptidyl peptidase IV in plasma and living systems. GP-BAN was designed on the basis of the catalytic properties and substrate preference of DPP-IV, and it could be readily hydrolyzed upon addition of DPP-IV under physiological conditions. Both reaction phenotyping and inhibition assays demonstrated that GP-BAN displayed good reactivity and high selectivity towards DPP-IV over other human serine hydrolases including FAP, DPP-VIII, and DPP-IX. The probe was successfully used to monitor the real activities of DPP-IV in complex biological systems including diluted plasma, while it could be used for high throughput screening of DPP-IV inhibitors by using human plasma or tissue preparations as enzyme sources. As a two-photon fluorescent probe, GP-BAN was also successfully used for two-photon imaging of endogenous DPP-IV in living cells and tissues, and showed high ratiometric imaging resolution and deep-tissue penetration ability. Taken together, a ratiometric two-photon fluorescent probe GP-BAN was developed and well-characterized for highly selective and sensitive detection of DPP-IV in complex biological systems, which could serve as a promising imaging tool to explore the biological functions and physiological roles of this key enzyme in living systems. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Control of photoemission delay in resonant two-photon transitions

    Science.gov (United States)

    Argenti, L.; Jiménez-Galán, Á.; Caillat, J.; Taïeb, R.; Maquet, A.; Martín, F.

    2017-04-01

    The photoelectron emission time delay τ associated with one-photon absorption, which coincides with half the Wigner delay τW experienced by an electron scattered off the ionic potential, is a fundamental descriptor of the photoelectric effect. Although it is hard to access directly from experiment, it is possible to infer it from the time delay of two-photon transitions, τ(2 ), measured with attosecond pump-probe schemes, provided that the contribution of the probe stage can be factored out. In the absence of resonances, τ can be expressed as the energy derivative of the one-photon ionization amplitude phase, τ =∂EargDE g , and, to a good approximation, τ =τ(2 )-τcc , where τcc is associated with the dipole transition between Coulomb functions. Here we show that, in the presence of a resonance, the correspondence between τ and ∂EargDE g is lost. Furthermore, while τ(2 ) can still be written as the energy derivative of the two-photon ionization amplitude phase, ∂EargDEg (2 ) , it does not have any scattering counterpart. Indeed, τ(2 ) can be much larger than the lifetime of an intermediate resonance in the two-photon process or more negative than the lower bound imposed on scattering delays by causality. Finally, we show that τ(2 ) is controlled by the frequency of the probe pulse, ωIR , so that by varying ωIR , it is possible to radically alter the photoelectron group delay.

  7. Two-Photon Excitation of Launched Cold Atoms in Flight

    Science.gov (United States)

    Goodsell, Anne; Gonzalez, Rene; Alejandro, Eduardo; Erwin, Emma

    2017-04-01

    We demonstrate two-photon bi-chromatic excitation of cold rubidium atoms in flight, using the pathway 5S1 / 2 -> 5P3 / 2 -> 5D5 / 2 with two resonant photons. In our experiment, atoms are laser-cooled in a magneto-optical trap and launched upward in discrete clouds with a controllable vertical speed of 7.1 +/-0.6 m/s and a velocity spread that is less than 10% of the launch speed. Outside the cooling beams, as high as 14 mm above the original center of the trap, the launched cold atoms are illuminated simultaneously by spatially-localized horizontal excitation beams at 780 nm (5S1 / 2 -> 5P3 / 2) and 776 nm (5P3 / 2 -> 5D5 / 2). We monitor transmission of the 780-nm beam over a range of intensities of 780-nm and 776-nm light. As the center of the moving cloud passes the excitation beams, we observe as much as 97.9 +/-1.2% transmission when the rate of two-photon absorption is high and the 5S1 / 2 and 5P3 / 2 states are depopulated, compared to 87.6 +/-0.9% transmission if only the 780-nm beam is present. This demonstrates two-photon excitation of a launched cold-atom source with controllable launch velocity and narrow velocity spread, as a foundation for three-photon excitation to Rydberg states. Research supported by Middlebury College Bicentennial Fund, Palen Fund, and Gladstone Award.

  8. Inclusive $D*^{+-}$ Production in Two-Photon Collisions at LEP

    CERN Document Server

    Achard, P.; Aguilar-Benitez, M.; Alcaraz, J.; Alemanni, G.; Allaby, J.; Aloisio, A.; Alviggi, M.G.; Anderhub, H.; Andreev, Valery 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.; Barillere, R.; Bartalini, P.; Basile, M.; Batalova, N.; Battiston, R.; Bay, A.; Becattini, F.; Becker, U.; Behner, F.; Bellucci, L.; Berbeco, R.; Berdugo, J.; Berges, P.; Bertucci, B.; Betev, B.L.; Biasini, M.; Biglietti, M.; Biland, A.; Blaising, J.J.; Blyth, S.C.; Bobbink, G.J.; Bohm, A.; Boldizsar, L.; Borgia, B.; Bottai, S.; Bourilkov, D.; Bourquin, M.; Braccini, S.; Branson, J.G.; Brochu, F.; Burger, J.D.; Burger, W.J.; Cai, X.D.; Capell, M.; Cara Romeo, G.; Carlino, G.; Cartacci, A.; Casaus, J.; Cavallari, F.; Cavallo, N.; Cecchi, C.; Cerrada, M.; Chamizo, M.; Chang, Y.H.; Chemarin, M.; Chen, A.; Chen, G.; Chen, G.M.; Chen, H.F.; Chen, H.S.; Chiefari, G.; Cifarelli, L.; Cindolo, F.; Clare, I.; Clare, R.; Coignet, G.; Colino, N.; Costantini, S.; de la Cruz, B.; Cucciarelli, S.; van Dalen, J.A.; de Asmundis, R.; Deglon, P.; Debreczeni, J.; Degre, A.; Deiters, K.; della Volpe, D.; Delmeire, E.; Denes, P.; DeNotaristefani, F.; De Salvo, A.; Diemoz, M.; Dierckxsens, M.; Dionisi, C.; Dittmar, M.; Doria, A.; Dova, M.T.; Duchesneau, D.; Echenard, B.; Eline, A.; El Mamouni, H.; Engler, A.; Eppling, F.J.; Ewers, A.; Extermann, P.; Falagan, M.A.; Falciano, S.; Favara, A.; Fay, J.; Fedin, O.; Felcini, M.; Ferguson, T.; Fesefeldt, H.; Fiandrini, E.; Field, J.H.; Filthaut, F.; Fisher, P.H.; Fisher, W.; Fisk, I.; Forconi, G.; Freudenreich, K.; Furetta, C.; Galaktionov, Iouri; Ganguli, S.N.; Garcia-Abia, Pablo; Gataullin, M.; Gentile, S.; Giagu, S.; Gong, Z.F.; Grenier, Gerald Jean; Grimm, O.; Gruenewald, M.W.; Guida, M.; van Gulik, R.; Gupta, V.K.; Gurtu, A.; Gutay, L.J.; Haas, D.; Hakobyan, R.S.; Hatzifotiadou, D.; Hebbeker, T.; Herve, Alain; Hirschfelder, J.; Hofer, H.; Hohlmann, M.; Holzner, G.; Hou, S.R.; Hu, Y.; Jin, B.N.; Jones, Lawrence W.; de Jong, P.; Josa-Mutuberria, I.; Kafer, D.; Kaur, M.; Kienzle-Focacci, M.N.; Kim, J.K.; Kirkby, Jasper; Kittel, W.; Klimentov, A.; Konig, A.C.; Kopal, M.; Koutsenko, V.; Kraber, M.; Kraemer, R.W.; Krenz, W.; Kruger, A.; Kunin, A.; Ladron 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.; Levtchenko, P.; Li, C.; Likhoded, S.; Lin, C.H.; Lin, W.T.; Linde, F.L.; Lista, L.; Liu, Z.A.; Lohmann, W.; Longo, E.; Lu, Y.S.; Lubelsmeyer, K.; Luci, C.; Luminari, L.; Lustermann, W.; Ma, W.G.; Malgeri, L.; Malinin, A.; Mana, C.; Mangeol, D.; 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.; Kluge, Hannelies; Ofierzynski, R.; Organtini, G.; Palomares, C.; Pandoulas, D.; Paolucci, P.; Paramatti, R.; Passaleva, G.; Patricelli, S.; Paul, Thomas Cantzon; Pauluzzi, M.; Paus, C.; Pauss, F.; Pedace, M.; Pensotti, S.; Perret-Gallix, D.; Petersen, B.; Piccolo, D.; Pierella, F.; Pioppi, M.; Piroue, P.A.; Pistolesi, E.; Plyaskin, V.; Pohl, M.; Pojidaev, V.; Pothier, J.; Prokofev, D.O.; Prokofiev, D.; Quartieri, J.; Rahal-Callot, G.; Rahaman, M.A.; Raics, P.; Raja, N.; Ramelli, R.; Rancoita, P.G.; Ranieri, R.; Raspereza, A.; Razis, P.; Ren, D.; Rescigno, M.; Reucroft, S.; Riemann, S.; Riles, Keith; Roe, B.P.; Romero, L.; Rosca, A.; Rosier-Lees, S.; Roth, Stefan; Rosenbleck, C.; Roux, B.; Rubio, J.A.; Ruggiero, G.; Rykaczewski, H.; Sakharov, A.; Saremi, S.; Sarkar, S.; Salicio, J.; Sanchez, E.; Sanders, M.P.; Schafer, C.; Schegelsky, V.; Schmidt-Kaerst, S.; Schmitz, D.; Schopper, H.; Schotanus, D.J.; Schwering, G.; Sciacca, C.; Servoli, L.; Shevchenko, S.; Shivarov, N.; Shoutko, V.; Shumilov, E.; Shvorob, A.; Siedenburg, T.; Son, D.; Spillantini, P.; Steuer, M.; Stickland, D.P.; Stoyanov, B.; Straessner, A.; Sudhakar, K.; Sultanov, G.; Sun, L.Z.; Sushkov, S.; Suter, H.; Swain, J.D.; Szillasi, Z.; Tang, X.W.; Tarjan, P.; Tauscher, L.; Taylor, L.; Tellili, B.; Teyssier, D.; Timmermans, Charles; Ting, Samuel C.C.; Ting, S.M.; Tonwar, S.C.; Toth, J.; Tully, C.; Tung, K.L.; Ulbricht, J.; Valente, E.; Van de Walle, R.T.; Veszpremi, V.; Vesztergombi, G.; Vetlitsky, I.; Vicinanza, D.; Viertel, G.; Villa, S.; Vivargent, M.; Vlachos, S.; Vodopianov, 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.; Zoller, 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.

  9. Transition to perturbative QCD in two-photon collisions

    OpenAIRE

    Hsieh, Ron-Chou; Li, Hsiang-nan

    2004-01-01

    We propose that the different angular distributions in two-photon collisions observed at low and high center-of-mass energies $W_{\\gamma\\gamma}$ indicate the transition from nonperturbative to perturbative QCD. We calculate the differential cross sections of $\\gamma \\gamma\\to\\pi \\pi$, $KK$ in the angle $\\theta$ of one of the final-state mesons using QCD sum rules and the perturbative QCD approach based on $k_T$ factorization theorem. Our predictions from sum rules (perturbative QCD) decrease ...

  10. Two-photon exclusive processes in quantum chromodynamics

    Energy Technology Data Exchange (ETDEWEB)

    Brodsky, S.J.

    1986-07-01

    QCD predictions for ..gamma gamma.. annihilation into single mesons, meson pairs, and baryon pairs are reviewed. Two-photon exclusive processes provide the most sensitive and practical measure of the distribution amplitudes, and thus a critical confrontation between QCD and experiment. Both the angular distribution and virtual photon mass dependence of these amplitudes are sensitive to the shapes of the phi (chi, Q). Novel effects involving the production of qq anti q anti q states at threshold are also discussed, and a new method is presented for systematically incorporating higher-order QCD corrections in ..gamma gamma.. reactions.

  11. Two-photon spectroscopy of excitons with entangled photons.

    Science.gov (United States)

    Schlawin, Frank; Mukamel, Shaul

    2013-12-28

    The utility of quantum light as a spectroscopic tool is demonstrated for frequency-dispersed pump-probe, integrated pump-probe, and two-photon fluorescence signals which show Ramsey fringes. Simulations of the frequency-dispersed transmission of a broadband pulse of entangled photons interacting with a three-level model of matter reveal how the non-classical time-bandwidth properties of entangled photons can be used to disentangle congested spectra, and reveal otherwise unresolved features. Quantum light effects are most pronounced at weak intensities when entangled photon pairs are well separated, and are gradually diminished at higher intensities when different photon pairs overlap.

  12. Two-photon mapping of localized field enhancements in thin nanostrip antennas

    DEFF Research Database (Denmark)

    Beermann, I.; Novikov, S.M.; Søndergaard, Thomas

    2008-01-01

    scanning optical microscopy, in which two-photon-excited photoluminescence (TPL) excited with a strongly focused laser beam at the wavelength 745 nm is detected. We use TPL images to map the local field enhancements from individual nanostrips at a resolution of 0.35µm and compare results with theoretical......Resonant scattering and local field enhancements by 11-nm-thin gold nanostrip antennas due to constructive interference of counter propagating slow surface plasmon polaritons is investigated. We characterize nanostrips of widths between 50-530 nm using both reflection spectroscopy and nonlinear...

  13. Near-Infrared Photoluminescent Polymer-Carbon Nanodots with Two-Photon Fluorescence.

    Science.gov (United States)

    Lu, Siyu; Sui, Laizhi; Liu, Junjun; Zhu, Shoujun; Chen, Anmin; Jin, Mingxing; Yang, Bai

    2017-04-01

    Near-infrared-emissive polymer-carbon nanodots (PCNDs) are fabricated by a newly developed facile, high-output strategy. The PCNDs emit at a wavelength of 710 nm with a quantum yield of 26.28%, which is promising for deep biological imaging and luminescent devices. Moreover, the PCNDs possess two-photon fluorescence; in vivo bioimaging and red-light-emitting diodes based on these PCNDs are demonstrated. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Two-photon probes for intracellular free metal ions, acidic vesicles, and lipid rafts in live tissues.

    Science.gov (United States)

    Kim, Hwan Myung; Cho, Bong Rae

    2009-07-21

    Optical imaging with fluorescence microscopy is a vital tool in the study of living systems. The most common method for cell imaging, one-photon microscopy (OPM), uses a single photon of higher energy to excite the fluorophore. However, two-photon microscopy (TPM), which uses two photons of lower energy as the excitation source, is growing in popularity among biologists because of several distinct advantages. Using TPM, researchers can image intact tissue for a long period of time with minimum interference from tissue preparation artifacts, self-absorption, autofluorescence, photobleaching, and photodamage. However, to make TPM a more versatile tool in biology, researchers need a wider variety of two-photon probes for specific applications. In this Account, we describe a series of two-photon probes that we developed that can visualize the distribution of intracellular metal ions, acidic vesicles, and lipid rafts in living cells and tissues. The development of these probes requires a significant two-photon cross section for the bright image and receptors (sensing moieties) that triggers the emission of the two-photon excited fluorescence upon binding with the ions or membrane in the living system. These probes also must be sensitive to the polarity of the environment to allow selective detection of cytosolic and membrane-bound probes. In addition, they need to be cell-permeable, water-soluble for the staining of cells and tissues, and highly photostable for long-term imaging. The resulting probes-AMg1 (Mg(2+)), ACa1-ACa3 (Ca(2+)), AZn1 and AZn2 (Zn(2+)), AH1, AH2, and AL1 (acidic vesicles), and CL2 (membrane)-use 2-acetyl-6-aminonaphthalene as the fluorophore and receptors for the target ions or membrane. All of these two-photon turn-on probes can detect the intracellular free metal ions, acidic vesicles, and lipid rafts at 100-300 microm depth in live tissues. Moreover, with ACa1-AM, we could simultaneously visualize the spontaneous Ca(2+) waves in the somas of

  15. Ultrafast responses of multi-branched compounds based on 1,3,5-triazine: investigation of the reason for enhanced two-photon absorption property

    Science.gov (United States)

    Wang, Yaochuan; Jiang, Yihua; Wang, Yizhuo; Wang, Guiqiu; Liu, Dajun; Hua, Jianli

    2017-08-01

    The fluorescence and ultrafast response of several modified tri-branched compounds based on 1,3,5-triazine ( T03-1, T03-2, and T03-3) were investigated to study the effects of stronger electron donor substitution or extended conjugated length on two-photon absorption/two-photon fluorescence and excited-state decay properties. After substitution with a stronger terminal electron donor, N, N-dimethylbenzenamine, the two-photon absorption cross section and fluorescence quantum yield of T03-2 were enhanced by 1.9-fold and 1.4-fold, respectively, compared with T03-1. After extending the conjugated length with thiophene groups connected to π-bridge styryl groups, a similar fluorescence quantum yield and larger two-photon absorption (2.3-fold) of T03-3 were obtained. The ultrafast response processes revealed the effects on the intramolecular charge-transfer properties. These results can also assist in the design of new molecular probes with improved two-photon absorption and two-photon fluorescence properties. Moreover, our study also indicates that these 1,3,5-triazine-based tri-branched derivatives can be employed as molecular probes for biological two-photon fluorescence imaging.

  16. 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...

  17. Optimization of targeted two-photon PDT triads for the treatment of head and neck cancers

    Science.gov (United States)

    Spangler, Charles W.; Starkey, Jean R.; Dubinina, Galyna; Fahlstrom, Carl; Shepard, Joyce

    2012-02-01

    Synthesis of new PDT triads that incorporate a tumor-killing porphyrin with large two-photon cross-section for 150 fs laser pulses (2000 GM) in the Near-infrared (NIR) at 840 nm, a NIR imaging agent, and a small peptide that targets over-expressed EGF receptors on the tumor surface. This triad formulation has been optimized over the past year to treat FADU Head and Neck SCC xenograft tumors in SCID mice. Effective PDT triad dose (1-10 mg/Kg) and laser operating parameters (840 nm, 15-45 min, 900 mW) have been established. Light, dark and PDT treatment toxicities were determined, showing no adverse effects. Previous experiments in phantom and mouse models indicate that tumors can be treated directly through the skin to effective depths between 2 and 5 cm. Treated mice demonstrated rapid tumor regression with some complete cures in as little as 15-20 days. No adverse effects were observed in any healthy tissue through which the focused laser beam passed before reaching the tumor site, and excellent healing occurred post treatment including rapid hair re-growth. Not all irradiation protocols lead to complete cures. Since two-photon PDT is carried out by rastering focused irradiation throughout the tumor, there is the possibility that as the treatment depth increases, some parts of the tumor may escape irradiation due to increased scattering, thus raising the possibility that tumor re-growth could be triggered by small islands of untreated cells, especially at the rapidly growing tumor margins, a problem we hope to alleviate by using image-guided two-photon PDT.

  18. All-optical histology using two photon laser scanning microscopy and ablation with ultrashort pulses

    Science.gov (United States)

    Tsai, Philbert S.

    This dissertation discusses the use of ultrashort laser pulses to image and manipulate tissue for the purpose of three-dimensional histological reconstruction of extended brain structures. Two photon laser scanning microscopy (TPLSM) and ultrashort pulsed laser ablation are used to provide in situ three-dimensional imaging through thick preparations of fixed tissue. Surface regions of fixed tissue are first imaged using TPLSM. The imaged regions are then removed by ablation with amplified, ultrashort laser pulses, thereby exposing a previously underlying tissue region for imaging. This process of imaging and ablation proceeds iteratively until the desired tissue volume has been processed. First, the principles, design, and construction of a two photon laser scanning microscope are discussed, followed by a discussion of the physical mechanisms of tissue ablation with ultrashort laser pulses. The compatibility of tissue ablation using ultrashort pulses with subsequent histological analysis, particularly with fluorescent microscopy, is evaluated. Tissue ablation with ultrashort laser pulses is found to produce ablated tissue surfaces that are smooth to within a micrometer. Intrinsic fluorescence as well as immunoreactivity are found to be resilient to the ablation process. The all-optical histological technique is demonstrated on brain tissue from rats and mice, including tissue from embryonic mouse as early at E15. The ablation process is shown to preserve both macroscopic and microscopic structures within tissue. To facilitate the all-optical histological analysis of neuronal vasculature and its relative distribution to surrounding neuronal tissue, a fluorescent gel perfusion technique is developed that provides a temperature-stabilized fluorescent label of the neuronal vasculature. The use of immunohistochemistry to label specific cell populations throughout an 800 micrometer-thick tissue section is demonstrated. Additionally, the immersion of fixed tissue in high

  19. Long-wavelength, photostable, two-photon excitable BODIPY fluorophores readily modifiable for molecular probes.

    Science.gov (United States)

    Zhang, Xinfu; Xiao, Yi; Qi, Jing; Qu, Junle; Kim, Bosung; Yue, Xiling; Belfield, Kevin D

    2013-09-20

    Near-infrared (NIR) fluorescent probes are increasingly popular in biological imaging and sensing, as long-wavelength (650-900 nm) excitation and emission have the advantages of minimum photodamage, deep tissue penetration, and minimum interference from autofluorescence in living systems. Here, a series of long-wavelength BODIPY dyes SPC, DC-SPC, DPC, and DC-DPC are synthesized conveniently and efficiently. They exhibit excellent photophysical properties in far red to near-infrared region, including large extinction coefficients, high fluorescence quantum yields, good photostability, and reasonable two-photon absorption cross section. Comparison of single-molecular imaging confirms that DPC is a much more efficient and more photostable NIR fluorophore than the commonly used Cy5. Also importantly, two kinds of convenient functionalization sites have been reserved: the aryl iodide for organometallic couplings and the terminal alkyne groups for click reactions. Further derivatives DC-SPC-PPh3 exhibit specificity to localize in mitochondria. The introduction of triphenylphosphonium (TPP) moieties mediates its hydrophilic-lipophilic balance and makes DC-SPC-PPh3 appropriate for cell labeling. Their long-wavelength emission at ∼650 nm can efficiently avoid the spectral crosstalk with other probes emitting in the visible light region. Superior photostability, low cytotoxicity, and two-photon excitable properties demonstrate its utility as a standard colocalizing agent to estimate the other probes' local distribution.

  20. Enhancement of two-photon photoluminescence and SERS for low-coverage gold films

    DEFF Research Database (Denmark)

    Novikov, Sergey M.; Beermann, Jonas; Frydendahl, Christian

    2016-01-01

    Electromagnetic field enhancement (FE) effects occurring in thin gold films 3-12-nm are investigated with two-photon photoluminescence (TPL) and Raman scanning optical microscopies. The samples are characterized using scanning electron microscopy images and linear optical spectroscopy. TPL images...... exhibit a strong increase in the level of TPL signals for films thicknesses 3-8-nm, near the percolation threshold. For some thicknesses, TPL measurements reveal super-cubic dependences on the incident power. We ascribe this feature to the occurrence of very strongly localized and enhanced electromagnetic...... fields due to multiple light scattering in random nanostructures that might eventually lead to white-light generation. Raman images exhibit increasing Raman signals when decreasing the film thickness from 12 to 6-nm and decreasing signal for the 3-nm-film. This feature correlates with the TPL...

  1. Two-photon Absorption In Quantum Dots,quantum Dashes And Related Materials

    Energy Technology Data Exchange (ETDEWEB)

    Jain, Ravinder

    2009-08-31

    We have proposed the use of USQDs for various deep-tissue biological imaging applications, notably wavelength-multiplexed multicolor imaging and intra-nuclear studies such as those involving cell apoptosis, and have studied the issue of maximizing two-photon absorption-induced fluorescence (TPAF) signals from CdSe/ZnS USQDs to be used for this application. In particular, using 2 nm USQDs, we have shown that the TPAF signal at 780 nm is ~ 8 times that at 850 nm and 68 times that at 900 nm, two wavelengths that have been used in previous studies using CdSe/ZnS SQDs for deep-tissue imaging of biological studies via TPAF .

  2. 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.

  3. 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

  4. Adaptive quantum control of two photon fluorescence on Coumarin 30 by using evolutionary algorithm

    Science.gov (United States)

    Poudel, Milan; Kolomenski, Alexender; Schuessler, Hans

    2006-10-01

    Two-photon excitation fluorescence of complex molecules (Coumarin-30) is successfully optimized by using feedback control pulse shaping technique. For such an optimization we have implemented an evolutionary algorithm [1], [2] in a Lab-view programming environment with a liquid crystal pulse shaper in a folded 4f set up. In the algorithm, one generation uses 48 individuals (vectors of voltage on the LC matrix).For each generation the fitness value is measured for every setting of the mask. A new generation is built from the previous by combining parents (the fittest individuals) and producing the desired degree of mutations (changes of the vector elements by some random value) to provide reasonable convergence. By successive repetition of this scheme, individuals corresponding to the highest fitness values will survive and produce offspring's for subsequent generations. Typically, convergence to the optimum value was achieved after 30 generations. Without any prior knowledge of the molecular system, the optimization goal was automatically achieved by changing the spectral phases [3]. The pulses before and after optimization were measured with GRENOUILLE, a type of second harmonic frequency resolved optical gating (SH FROG). To find the efficient pulse with lower intensity, three types of optimization were performed, the two photon fluorescence signal, the second harmonic signal and the ratio between them. The Intensity of two photon fluorescence of coumarin-30 could be increased noticeably compared to the transform limited pulse optimizing the second harmonic generation. The experimental results appear to be the potential applications of coherent control to the complicated molecular system as well as in bio medical imaging.

  5. Two-photon excitation of rubidium atoms inside porous glass

    Science.gov (United States)

    Amy, L.; Lenci, L.; Villalba, S.; Failache, H.; Lezama, A.

    2017-10-01

    We study the two-photon laser excitation to the 5 D5 /2 energy level of 85Rb atoms contained in the interstices of a porous material made from sintered ground glass with typical pore dimensions in the 10-100 μ m range. The excitation spectra show unusual flat-top line shapes, which are shown to be the consequence of wave-vector randomization of the laser light in the porous material. For large atomic densities, the spectra are affected by radiation trapping around the D2 transitions. The effect of the transient atomic response limited by the time of flight between pores walls appears to have a minor influence in the excitation spectra. It is however revealed by the shortening of the temporal evolution of the emitted blue light following a sudden switch-off of the laser excitation.

  6. 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.

  7. High-dynamic-range cationic two-photon photopolymerization

    Science.gov (United States)

    Boiko, Yuri B.; Costa, Joannes M.; Wang, Mark M.; Esener, Sadik C.

    2001-06-01

    Cationic-induced two-photon photopolymerization is demonstrated at 710 nm, using an isopropylthioxanthone/diarylidonium salt initiating system for the cationic polymerization of an epoxide. The polymerization threshold J2th is found to be approximately 1 GW/cm2, with a dynamic range of > 100, i.e. the material can be fully polymerized at intensities > 100 times the threshold level without damage. The polymerization rate R is found to be proportional to the m equals 1.7 power of the intensity, or R equals [C (J-J2th)]m equals [C (J-J2th)]1.7, which implies a significantly stronger localization of the photochemical response than that of free radical photoinitiators. R and J2th significantly improve when the concentration z of the initiator (onium salt) increases.

  8. Results from the OLYMPUS Two-Photon Exchange Experiment

    Science.gov (United States)

    O'Connor, Colton; Olympus Collaboration

    2017-01-01

    Measurements of the proton's electric-to-magnetic form factor ratio obtained by different methods disagree significantly in a way that depends on Q2. The OLYMPUS experiment was designed to empirically quantify two-photon exchange in lepton-proton scattering, an effect that, in some models, can fully account for this disagreement. This was achieved at the DORIS storage ring at DESY by measuring the ratio of the elastic cross-sections for positron-proton and electron-proton scattering with alternating 2.01 GeV lepton beams incident on an internal hydrogen gas target. Data were collected with an integrated luminosity of over 4.0 fb-1 using a large-acceptance toroidal spectrometer and multiple luminosity monitoring systems, allowing for precise results (work is supported by DOE Grant DE-FG02-94ER40818.

  9. Two-photon quantum walk in a multimode fiber

    Science.gov (United States)

    Defienne, Hugo; Barbieri, Marco; Walmsley, Ian A.; Smith, Brian J.; Gigan, Sylvain

    2016-01-01

    Multiphoton propagation in connected structures—a quantum walk—offers the potential of simulating complex physical systems and provides a route to universal quantum computation. Increasing the complexity of quantum photonic networks where the walk occurs is essential for many applications. We implement a quantum walk of indistinguishable photon pairs in a multimode fiber supporting 380 modes. Using wavefront shaping, we control the propagation of the two-photon state through the fiber in which all modes are coupled. Excitation of arbitrary output modes of the system is realized by controlling classical and quantum interferences. This report demonstrates a highly multimode platform for multiphoton interference experiments and provides a powerful method to program a general high-dimensional multiport optical circuit. This work paves the way for the next generation of photonic devices for quantum simulation, computing, and communication. PMID:27152325

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

    Directory of Open Access Journals (Sweden)

    Denis Soulet

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

  11. Brain refractive index measured in vivo with high-NA defocus-corrected full-field OCT and consequences for two-photon microscopy.

    Science.gov (United States)

    Binding, Jonas; Ben Arous, Juliette; Léger, Jean-François; Gigan, Sylvain; Boccara, Claude; Bourdieu, Laurent

    2011-03-14

    Two-photon laser scanning microscopy (2PLSM) is an important tool for in vivo tissue imaging with sub-cellular resolution, but the penetration depth of current systems is potentially limited by sample-induced optical aberrations. To quantify these, we measured the refractive index n' in the somatosensory cortex of 7 rats in vivo using defocus optimization in full-field optical coherence tomography (ff-OCT). We found n' to be independent of imaging depth or rat age. From these measurements, we calculated that two-photon imaging beyond 200 µm into the cortex is limited by spherical aberration, indicating that adaptive optics will improve imaging depth.

  12. Confinement of pyridinium hemicyanine dye within an anionic metal-organic framework for two-photon-pumped lasing

    Science.gov (United States)

    Yu, Jiancan; Cui, Yuanjing; Xu, Hui; Yang, Yu; Wang, Zhiyu; Chen, Banglin; Qian, Guodong

    2013-10-01

    Two-photon-pumped dye lasers are very important because of their applications in wavelength up-conversion, optical data storage, biological imaging and photodynamic therapy. Such lasers are very difficult to realize in the solid state because of the aggregation-caused quenching. Here we demonstrate a new two-photon-pumped micro-laser by encapsulating the cationic pyridinium hemicyanine dye into an anionic metal-organic framework (MOF). The resultant MOF⊃dye composite exhibits significant two-photon fluorescence because of the large absorption cross-section and the encapsulation-enhanced luminescent efficiency of the dye. Furthermore, the well-faceted MOF crystal serves as a natural Fabry-Perot resonance cavity, leading to lasing around 640 nm when pumped with a 1064-nm pulse laser. This strategy not only combines the crystalline benefit of MOFs and luminescent behaviour of organic dyes but also creates a new synergistic two-photon-pumped lasing functionality, opening a new avenue for the future creation of solid-state photonic materials and devices.

  13. Targeted two-photon photodynamic therapy for the treatment of subcutaneous tumors

    Science.gov (United States)

    Spangler, Charles W.; Starkey, Jean R.; Meng, Fanqing; Gong, Aijun; Drobizhev, Mikhail; Rebane, Aleksander; Moss, B.

    2005-04-01

    Photodynamic therapy (PDT) has developed into a mature technology over the past several years, and is currently being exploited for the treatment of a variety of cancerous tumors, and more recently for age-related wet macular degeneration of the eye. However, there are still some unresolved problems with PDT that are retarding a more general acceptance in clinical settings, and thus, for the most part, the treatment of most cancerous rumors still involves some combination of invasive surgery, chemotherapy and radiation treatment, particularly subcutaneous tumors. Currently approved PDT agents are activated in the Visible portion of the spectrum below 700 nm, Laser light in this spectral region cannot penetrate the skin more than a few millimeters, and it would be more desirable if PDT could be initiated deep in the Near-infrared (NIR) in the tissue transparency window (700-1000 nm). MPA Technologies, Inc. and Rasiris, Inc. have been co-developing new porphyrin PDT designed to have greatly enhanced intrinsic two-photon cross-sections (>800 GM units) whose two-photon absorption maxima lie deep in the tissue transparency window (ca. 780-850 nm), and have solubility characteristics that would allow for direct IV injection into animal models. Classical PDT also suffers from the lengthy time necessary for accumulation at the tumor site, a relative lack of discrimination between healthy and diseased tissue, particularly at the tumor margins, and difficulty in clearing from the system in a reasonable amount of time post-PDT. We have recently discovered a new design paradigm for the delivery of our two-photon activated PDT agents by incorporating the porphyrins into a triad ensemble that includes a small molecule targeting agent that directs the triad to over-expressed tumor receptor sites, and a NIR one-photon imaging agent that allows the tracking of the triad in terms of accumulation and clearance rates. We are currently using these new two-photon PDT triads in efficacy

  14. Sensitive and rapid detection of endogenous hydrogen sulfide distributing in different mouse viscera via a two-photon fluorescent probe

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Qian [State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082 (China); Yang, Jinfeng [Tumor Hospital, Xiangya School of Medicine, Central South University, Changsha, 410013 (China); Li, Yinhui, E-mail: yinhuili16@163.com [State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082 (China); Zheng, Jing [State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082 (China); Yang, Ronghua, E-mail: yangrh@pku.edu.cn [State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082 (China); School of Chemistry and Biological Engineering, Changsha University of Science and Technology, Changsha, 410004 (China)

    2015-10-08

    Development of efficient methods for detection of endogenous H{sub 2}S in living cells and tissues is of considerable significance for better understanding the biological and pathological functions of H{sub 2}S. Two-photon (TP) fluorescent probes are favorable as powerful molecular tools for studying physiological process due to its non-invasiveness, high spatiotemporal resolution and deep-tissues imaging. Up to date, several TP probes for intracellular H{sub 2}S imaging have been designed, but real-time imaging of endogenous H{sub 2}S-related biological processes in tissues is hampered due to low sensitivity, long response time and interference from other biothiols. To address this issue, we herein report a novel two-photon fluorescent probe (TPP-H{sub 2}S) for highly sensitive and fast monitoring and imaging H{sub 2}S levels in living cells and tissues. In the presence of H{sub 2}S, it exhibits obviously improved sensitivity (LOD: 0.12 μM) and fast response time (about 2 min) compared with the reported two-photon H{sub 2}S probes. With two-photon excitation, TPP-H{sub 2}S displays high signal-to-noise ratio and sensitivity even no interference in cell growth media. As further application, TPP-H{sub 2}S is applied for fast imaging of H{sub 2}S in living cells and different fresh tissues by two-photon confocal microscope. Most importantly we first measured the endogenous H{sub 2}S level in different viscera by vivisection and found that the distribution of endogenous H{sub 2}S mostly in brain, liver and lung. The excellent sensing properties of TPP-H{sub 2}S make it a practically useful tool for further studying biological roles of H{sub 2}S. - Highlights: • This two-photon probe exhibits an improved sensitivity and response time to H{sub 2}S. • This probe shows excellent membrane permeability and fast visualization of H{sub 2}S in living cells and tissues. • This probe is successfully applied to measure the endogenously produced H{sub 2}S levels in

  15. Microspot two-photon photoemission spectroscopy for CuPc film on HOPG

    Energy Technology Data Exchange (ETDEWEB)

    Yamada, T.; Yamamoto, R.; Munakata, T., E-mail: munakata@chem.sci.osaka-u.ac.jp

    2015-10-01

    Highlights: • Unoccupied levels of CuPc/HOPG are assigned by using 2PPE microspectroscopy. • Lateral distribution of unoccupied energy levels is imaged. • Modified IPS stabilized by the hole localized in the 2nd layer molecule is identified. - Abstract: Microspot two-photon photoemission (micro-2PPE) spectroscopy has been applied to measure the lateral distribution of unoccupied levels on copper phthalocyanine (CuPc) film on HOPG. In addition to the LUMO-derived level and the image potential state (IPS) on the film, we identified the modified IPS which is stabilized by the hole localized in a molecule. We show that modified IPS is observed only on bilayer area, reflecting the localization of the hole in a molecule. The modified IPS is absent on monolayer area, because the hole strongly interacts with substrate.

  16. A coumarin-based two-photon probe for hydrogen peroxide.

    Science.gov (United States)

    Zhang, Kai-Ming; Dou, Wei; Li, Peng-Xuan; Shen, Rong; Ru, Jia-Xi; Liu, Wei; Cui, Yu-Mei; Chen, Chun-Yang; Liu, Wei-Sheng; Bai, De-Cheng

    2015-02-15

    A new fluorescence probe was developed for hydrogen peroxide (H2O2) detection based on donor-excited photo induced electron transfer (D-PET) mechanism, together with the benzil as a quenching and recognizing moiety. The benzil could convert to benzoic anhydride via a Baeyer-Villiger type reaction in the presence of H2O2, followed by hydrolysis of benzoicanhydride to give benzoic acid, and the fluorophore released. The probe was synthesized by a 6-step procedure starting from 4-(diethylamino)salicylaldehyde. A density functional theory (DFT) calculation was performed to demonstrate that the benzil was a fluorescence quencher. The probe was evaluated in both one-photon and two-photon mode, and it exhibited high selectivity toward H2O2 over other reactive oxygen species and high sensitivity with a detection limit of 0.09 μM. Furthermore, the probe was successfully applied to cell imaging of intracellular H2O2 levels with one-photon microscopy and two-photon microscopy. The superior properties of the probe made it of great potential use in more chemical and biological researches. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Two-photon-like microscopy with orders-of-magnitude lower illumination intensity via two-step fluorescence.

    Science.gov (United States)

    Ingaramo, Maria; York, Andrew G; Andrade, Eric J; Rainey, Kristin; Patterson, George H

    2015-09-03

    We describe two-step fluorescence microscopy, a new approach to non-linear imaging based on positive reversible photoswitchable fluorescent probes. The protein Padron approximates ideal two-step fluorescent behaviour: it equilibrates to an inactive state, converts to an active state under blue light, and blue light also excites this active state to fluoresce. Both activation and excitation are linear processes, but the total fluorescent signal is quadratic, proportional to the square of the illumination dose. Here, we use Padron's quadratic non-linearity to demonstrate the principle of two-step microscopy, similar in principle to two-photon microscopy but with orders-of-magnitude better cross-section. As with two-photon, quadratic non-linearity from two-step fluorescence improves resolution and reduces unwanted out-of-focus excitation, and is compatible with structured illumination microscopy. We also show two-step and two-photon imaging can be combined to give quartic non-linearity, further improving imaging in challenging samples. With further improvements, two-step fluorophores could replace conventional fluorophores for many imaging applications.

  18. Theoretical Studies on Two-Photon Fluorescent Hg2+ Probes Based on the Coumarin-Rhodamine System

    Directory of Open Access Journals (Sweden)

    Yujin Zhang

    2017-07-01

    Full Text Available The development of fluorescent sensors for Hg2+ has attracted much attention due to the well-known adverse effects of mercury on biological health. In the present work, the optical properties of two newly-synthesized Hg2+ chemosensors based on the coumarin-rhodamine system (named Pro1 and Pro2 were systematically investigated using time-dependent density functional theory. It is shown that Pro1 and Pro2 are effective ratiometric fluorescent Hg2+ probes, which recognize Hg2+ by Förster resonance energy transfer and through bond energy transfer mechanisms, respectively. To further understand the mechanisms of the two probes, we have developed an approach to predict the energy transfer rate between the donor and acceptor. Using this approach, it can be inferred that Pro1 has a six times higher energy transfer rate than Pro2. Thus the influence of spacer group between the donor and acceptor on the sensing performance of the probe is demonstrated. Specifically, two-photon absorption properties of these two probes are calculated. We have found that both probes show significant two-photon responses in the near-infrared light region. However, only the maximum two-photon absorption cross section of Pro1 is greatly enhanced with the presence of Hg2+, indicating that Pro1 can act as a potential two-photon excited fluorescent probe for Hg2+. The theoretical investigations would be helpful to build a relationship between the structure and the optical properties of the probes, providing information on the design of efficient two-photon fluorescent sensors that can be used for biological imaging of Hg2+ in vivo.

  19. Multi-color femtosecond source for simultaneous excitation of multiple fluorescent proteins in two-photon fluorescence microscopy

    Science.gov (United States)

    Wang, Ke; Liu, Tzu-Ming; Wu, Juwell; Horton, Nicholas G.; Lin, Charles P.; Xu, Chris

    2013-02-01

    Simultaneous imaging of cells expressing multiple fluorescent proteins (FPs) is of particular interest in applications such as mapping neural circuits, tracking multiple immune cell populations, etc. To visualize both in vivo and ex vivo tissue morphology and physiology at a cellular level deep within scattering tissues, two-photon fluorescence microscopy (2PM) is a powerful tool that has found wide applications. However, simultaneous imaging of multiple FPs with 2PM is greatly hampered by the lack of proper ultrafast lasers offering multi-color femtosecond pulses, each targeting the two-photon absorption peak of a different FP. Here we demonstrate simultaneous two-photon fluorescence excitation of RFP, YFP, and CFP in human melanoma cells engineered to express a "rainbow" pallet of colors, using a novel fiber-based source with energetic, three-color femtosecond pulses. The three-color pulses, centered at 775 nm, 864 nm and 950 nm, are obtained through second harmonic generation of the 1550 nm pump laser and SHG of the solitons at 1728 nm and 1900 nm generated through soliton self-frequency shift (SSFS) of the pump laser in a large-mode-area (LMA) fiber. The resulting wavelengths are well matched to the two-photon absorption peaks of the three FPs for efficient excitation. Our results demonstrate that multi-color femtosecond pulse generation using SSFS and a turn-key, fiber-based femtosecond laser can fulfill the requirements for simultaneous imaging of multiple FPs in 2PM, opening new opportunities for a wide range of biological applications where non-invasive, high-resolution imaging of multiple fluorescent indicators is required.

  20. 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.

  1. 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.

  2. Improving the fidelity of two-photon absorption reference standards

    Science.gov (United States)

    de Reguardati, S.; Pahapill, J.; Rammo, M.; Rebane, A.

    2017-02-01

    Reference standards with well-defined femtosecond two-photon absorption (2PA) properties facilitate accurate measurement of nonlinear-optical spectra by bypassing tedious characterization of the photon flux. The 2PA standards are increasingly used for developing advanced multi-photon fluorescent probes and, since recently, also for probing intra- and intermolecular electrostatic interactions. We have recently reported 2PA cross section values of a set of common organic dyes in different solvents in 680-1050 nm wavelength range with estimated accuracy of 8%. In the present work, we aim at further improving the accuracy and fidelity of the absolute 2PA cross section data by comparing in a pair-wise manner the relative 2PA efficiency of nine standards with partially overlapping absorption- and fluorescence emission spectra. We measure the relative 2PA-induced fluorescence for each pair under identical excitation conditions, which allows revealing inconsistencies potentially present in the previously published data due to errors in estimating the excitation laser beam spatial- and temporal profile, pulse energy and other critical parameters. Our current measurements confirmed and in some cases improved previously reported error margins thus improving the fidelity of the reference data. We also present refined 2PA cross section data on 9-Chloroanthracene in dichloromethane.

  3. Coherent two-photon excitation of quantum dots

    Science.gov (United States)

    Ostermann, L.; Huber, T.; Prilmüller, M.; Solomon, G. S.; Ritsch, H.; Weihs, G.; Predojević, A.

    2016-04-01

    Single semiconductor quantum dots, due to their discrete energy structure, form single photon and twin photon sources that are characterized by a well-defined frequency of the emitted photons and inherently sub-Poissonian statistics. The single photons are generated through a recombination of an electron-hole pair formed by an electron from the conduction band and a hole from the valence band. When excited to the biexciton state quantum dots can provide pairs of photons emitted in a cascade. It has been shown that this biexciton-exciton cascade can deliver entangled pairs of photons. To achieve a deterministic generation of photon pairs from a quantum dot system one requires exciting it using a two-photon resonant excitation of the biexciton. Particularly, an efficient and coherent excitation of the biexciton requires the elimination of the single exciton probability amplitude in the excitation pulse and reaching the lowest possible degree of dephasing caused by the laser excitation. These two conditions impose contradictory demands on the excitation pulse-length and its intensity. We addressed this problem from a point of view that does not include interaction of the quantum dot with the semiconductor environment. We found an optimized operation regime for the system under consideration and provide guidelines on how to extend this study to other similar systems. In particular, our study shows that an optimal excitation process requires a trade-off between the biexciton binding energy and the excitation laser pulse length.

  4. 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.

  5. Cationic two-photon induced polymerization with high dynamic range

    Science.gov (United States)

    Boiko, Yuri B.; Costa, Joannes; Wang, Mark M.; Esener, Sadik C.

    2001-05-01

    Cationic-induced two-photon photo-polymerization is demonstrated at 710 nm, using an isopropylthioxanthone / diarylidonium salt initiating system for the cationic polymerization of an epoxide. In-situ monitoring of the polymer conversion using interferometry allows for determination of the polymerization threshold J2th, polymerization rate R and its dependence of initiator's concentration z. Best J2th achieved is 1 GW/cm 2 , with a dynamic range of > 100, i.e. the material can be fully polymerized at intensities > 100 times the threshold level without damage. The R is found to be proportional to the m=1.7 power of the intensity, or R =[C(J-J2th)]m =[C(J-J2th)]1.7 , which implies a significantly stronger localization of the photochemical response than that of free radical photoinitiators. Both R and J2th significantly improve when the concentration z of the initiator (onium salt) increases, reduction of J2th exhibiting z -m trend.

  6. 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.

  7. Two-photon interference of polarization-entangled photons in a Franson interferometer.

    Science.gov (United States)

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

    2017-07-18

    We present two-photon interference experiments with polarization-entangled photon pairs in a polarization-based Franson-type interferometer. Although the two photons do not meet at a common beamsplitter, a phase-insensitive Hong-Ou-Mandel type two-photon interference peak and dip fringes are observed, resulting from the two-photon interference effect between two indistinguishable two-photon probability amplitudes leading to a coincidence detection. A spatial quantum beating fringe is also measured for nondegenerate photon pairs in the same interferometer, although the two-photon states have no frequency entanglement. When unentangled polarization-correlated photons are used as an input state, the polarization entanglement is successfully recovered through the interferometer via delayed compensation.

  8. CdSe/CdS-quantum rods: fluorescent probes for in vivo two-photon laser scanning microscopy

    Science.gov (United States)

    Dimitrijevic, Jelena; Krapf, Lisa; Wolter, Christopher; Schmidtke, Christian; Merkl, Jan-Philip; Jochum, Tobias; Kornowski, Andreas; Schüth, Anna; Gebert, Andreas; Hüttmann, Gereon; Vossmeyer, Tobias; Weller, Horst

    2014-08-01

    CdSe/CdS-Quantum-dots-quantum-rods (QDQRs) with an aspect ratio of ~6 are prepared via the seeded growth method, encapsulated within a shell of crosslinked poly(isoprene)-block-poly(ethylene glycol) (PI-b-PEG) diblock copolymer, and transferred from the organic phase into aqueous media. Their photoluminescence quantum yield (PLQY) of 78% is not compromised by the phase transfer. Within a period of two months the PLQY of QDQRs in aqueous solution at neutral pH decreases only slightly (to ~65%). The two-photon (TP) action cross sections of QDQRs (~105 GM) are two orders of magnitude higher than those of CdSe/CdS/ZnS-core/shell/shell quantum dots (QDs, ~103 GM) with comparable diameter (~5 nm). After applying PI-b-PEG encapsulated QDQRs onto the small intestinal mucosa of mice in vivo, their strong red fluorescence can easily be observed by two-photon laser scanning microscopy (TPLSM) and clearly distinguished from autofluorescent background. Our results demonstrate that PI-b-PEG encapsulated CdSe/CdS-QDQRs are excellent probes for studying the uptake and fate of nanoparticles by two-photon imaging techniques in vivo.CdSe/CdS-Quantum-dots-quantum-rods (QDQRs) with an aspect ratio of ~6 are prepared via the seeded growth method, encapsulated within a shell of crosslinked poly(isoprene)-block-poly(ethylene glycol) (PI-b-PEG) diblock copolymer, and transferred from the organic phase into aqueous media. Their photoluminescence quantum yield (PLQY) of 78% is not compromised by the phase transfer. Within a period of two months the PLQY of QDQRs in aqueous solution at neutral pH decreases only slightly (to ~65%). The two-photon (TP) action cross sections of QDQRs (~105 GM) are two orders of magnitude higher than those of CdSe/CdS/ZnS-core/shell/shell quantum dots (QDs, ~103 GM) with comparable diameter (~5 nm). After applying PI-b-PEG encapsulated QDQRs onto the small intestinal mucosa of mice in vivo, their strong red fluorescence can easily be observed by two-photon laser

  9. Coherent anti-Stokes Raman scattering and two photon excited fluorescence for neurosurgery.

    Science.gov (United States)

    Romeike, Bernd F M; Meyer, Tobias; Reichart, Rupert; Kalff, Rolf; Petersen, Iver; Dietzek, Benjamin; Popp, Jürgen

    2015-04-01

    There is no established method for in vivo imaging during biopsy and surgery of the brain, which is capable to generate competitive images in terms of resolution and contrast comparable with histopathological staining. Coherent anti-Stokes Raman scattering (CARS) and two photon excited fluorescence (TPEF) microscopy are non-invasive all optical imaging techniques that are capable of high resolution, label-free, real-time, nondestructive examination of living cells and tissues. They provide image contrast based on the molecular composition of the specimen which allows the study of large tissue areas of frozen tissue sections ex vivo. Here, preliminary data on 55 lesions of the central nervous system are presented. The generated images very nicely demonstrate cytological and architectural features required for pathological tumor typing and grading. Furthermore, information on the molecular content of a probe is provided. The tool will be implemented into a biopsy needle or endoscope in the near future for in vivo studies. With this promising multimodal imaging approach the neurosurgeon might directly see blood vessels to minimize the risk for biopsy associated hemorrhages. The attending neuropathologist might directly identify the tumor and guide the selection of representative specimens for further studies. Thus, collection of non-representative material could be avoided and the risk to injure eloquent brain tissue minimized. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. Design and performance of an ultra-flexible two-photon microscope for in vivo research

    Science.gov (United States)

    Mayrhofer, Johannes M.; Haiss, Florent; Haenni, Dominik; Weber, Stefan; Zuend, Marc; Barrett, Matthew J. P.; Ferrari, Kim David; Maechler, Philipp; Saab, Aiman S.; Stobart, Jillian L.; Wyss, Matthias T.; Johannssen, Helge; Osswald, Harald; Palmer, Lucy M.; Revol, Vincent; Schuh, Claus-Dieter; Urban, Claus; Hall, Andrew; Larkum, Matthew E.; Rutz-Innerhofer, Edith; Zeilhofer, Hanns Ulrich; Ziegler, Urs; Weber, Bruno

    2015-01-01

    We present a cost-effective in vivo two-photon microscope with a highly flexible frontend for in vivo research. Our design ensures fast and reproducible access to the area of interest, including rotation of imaging plane, and maximizes space for auxiliary experimental equipment in the vicinity of the animal. Mechanical flexibility is achieved with large motorized linear stages that move the objective in the X, Y, and Z directions up to 130 mm. 360° rotation of the frontend (rotational freedom for one axis) is achieved with the combination of a motorized high precision bearing and gearing. Additionally, the modular design of the frontend, based on commercially available optomechanical parts, allows straightforward updates to future scanning technologies. The design exceeds the mobility of previous movable microscope designs while maintaining high optical performance. PMID:26600989

  11. 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.)

  12. An aggregation-induced emission luminophore with multi-stimuli single- and two-photon fluorescence switching and large two-photon absorption cross section.

    Science.gov (United States)

    Xu, Bingjia; Xie, Mingyuan; He, Jiajun; Xu, Bin; Chi, Zhenguo; Tian, Wenjing; Jiang, Long; Zhao, Fuli; Liu, Siwei; Zhang, Yi; Xu, Zhizhan; Xu, Jiarui

    2013-01-11

    A novel aggregation- and crystallization-induced emission luminophore (ENPOMe) containing tetraphenylethene and acrylonitrile moieties with high fluorescence efficiency (Φ(F) of up to 0.85) has been easily synthesized. ENPOMe has an exceptionally large two-photon absorption cross section (σ) of 5548 GM, and exhibits striking multi-stimuli-responsive single- and two-photon fluorescence switching with excellent reversibility in the solid state.

  13. A High Performance, Cost-Effective, Open-Source Microscope for Scanning Two-Photon Microscopy that Is Modular and Readily Adaptable

    Science.gov (United States)

    Rosenegger, David G.; Tran, Cam Ha T.; LeDue, Jeffery; Zhou, Ning; Gordon, Grant R.

    2014-01-01

    Two-photon laser scanning microscopy has revolutionized the ability to delineate cellular and physiological function in acutely isolated tissue and in vivo. However, there exist barriers for many laboratories to acquire two-photon microscopes. Additionally, if owned, typical systems are difficult to modify to rapidly evolving methodologies. A potential solution to these problems is to enable scientists to build their own high-performance and adaptable system by overcoming a resource insufficiency. Here we present a detailed hardware resource and protocol for building an upright, highly modular and adaptable two-photon laser scanning fluorescence microscope that can be used for in vitro or in vivo applications. The microscope is comprised of high-end componentry on a skeleton of off-the-shelf compatible opto-mechanical parts. The dedicated design enabled imaging depths close to 1 mm into mouse brain tissue and a signal-to-noise ratio that exceeded all commercial two-photon systems tested. In addition to a detailed parts list, instructions for assembly, testing and troubleshooting, our plan includes complete three dimensional computer models that greatly reduce the knowledge base required for the non-expert user. This open-source resource lowers barriers in order to equip more laboratories with high-performance two-photon imaging and to help progress our understanding of the cellular and physiological function of living systems. PMID:25333934

  14. A high performance, cost-effective, open-source microscope for scanning two-photon microscopy that is modular and readily adaptable.

    Directory of Open Access Journals (Sweden)

    David G Rosenegger

    Full Text Available Two-photon laser scanning microscopy has revolutionized the ability to delineate cellular and physiological function in acutely isolated tissue and in vivo. However, there exist barriers for many laboratories to acquire two-photon microscopes. Additionally, if owned, typical systems are difficult to modify to rapidly evolving methodologies. A potential solution to these problems is to enable scientists to build their own high-performance and adaptable system by overcoming a resource insufficiency. Here we present a detailed hardware resource and protocol for building an upright, highly modular and adaptable two-photon laser scanning fluorescence microscope that can be used for in vitro or in vivo applications. The microscope is comprised of high-end componentry on a skeleton of off-the-shelf compatible opto-mechanical parts. The dedicated design enabled imaging depths close to 1 mm into mouse brain tissue and a signal-to-noise ratio that exceeded all commercial two-photon systems tested. In addition to a detailed parts list, instructions for assembly, testing and troubleshooting, our plan includes complete three dimensional computer models that greatly reduce the knowledge base required for the non-expert user. This open-source resource lowers barriers in order to equip more laboratories with high-performance two-photon imaging and to help progress our understanding of the cellular and physiological function of living systems.

  15. Two-Photon Microscopy Analysis of Gold Nanoparticle Uptake in 3D Cell Spheroids.

    Directory of Open Access Journals (Sweden)

    Tushar D Rane

    Full Text Available Nanomaterials can be synthesized from a wide range of material systems in numerous morphologies, creating an extremely diverse portfolio. As result of this tunability, these materials are emerging as a new class of nanotherapeutics and imaging agents. One particularly interesting nanomaterial is the gold nanoparticle. Due to its inherent biocompatibility and tunable photothermal behavior, it has made a rapid transition from the lab setting to in vivo testing. In most nanotherapeutic applications, the efficacy of the agent is directly related to the target of interest. However, the optimization of the AuNP size and shape for efficacy in vitro, prior to testing in in vivo models of a disease, has been largely limited to two dimensional monolayers of cells. Two dimensional cell cultures are unable to reproduce conditions experienced by AuNP in the body. In this article, we systematically investigate the effect of different properties of AuNP on the penetration depth into 3D cell spheroids using two-photon microscopy. The 3D spheroids are formed from the HCT116 cell line, a colorectal carcinoma cell line. In addition to studying different sizes and shapes of AuNPs, we also study the effect of an oligo surface chemistry. There is a significant difference between AuNP uptake profiles in the 2D monolayers of cells as compared to the 3D cell spheroids. Additionally, the range of sizes and shapes studied here also exhibit marked differences in uptake penetration depth and efficacy. Finally, our results demonstrate that two-photon microscopy enables quantitative AuNP localization and concentration data to be obtained at the single spheroid level without fluorescent labeling of the AuNP, thus, providing a viable technique for large scale screening of AuNP properties in 3D cell spheroids as compared to tedious and time consuming techniques like electron microscopy.

  16. Quantum correlation of path-entangled two-photon states in waveguide arrays with defects

    Directory of Open Access Journals (Sweden)

    Yiling Dou

    2014-04-01

    Full Text Available We study the quantum correlation of path-entangled states of two photons in coupled one-dimensional waveguide arrays with lattice defects. Both off-diagonal and diagonal defects are considered, which show different effects on the quantum correlation of path-entangled two-photon states. Two-photon bunching or anti-bunching effects can be observed and controlled. The two photons are found to have a tendency to bunch at the side lobes with a repulsive off-diagonal defect, and the path-entanglement of the input two-photon state can be preserved during the propagation. We also found that defect modes may play an important role on the two-photon correlation of path-entangled states in the waveguide arrays. Due to the quantum interference effect, intriguing evolution dynamics of the two-photon correlation matrix elements with oscillation frequencies being either twice of or the same as that of a classical light wave, depending on the position of the correlation matrix element, is observed. Our results show that it is possible to manipulate the two-photon correlation properties of path-entangled states in waveguide arrays with lattice defects.

  17. Characterization of gradient-index lens-fiber spacing toward applications in two-photon fluorescence endoscopy

    Science.gov (United States)

    Fu, Ling; Gan, Xiaosong; Gu, Min

    2005-12-01

    We report on the experimental investigation into the characterization of two-photon fluorescence microscopy based on the separation distance of a single-mode optical fiber coupler and a gradient-index (GRIN) rod lens. The collected two-photon fluorescence signal exhibits a maximum intensity at a defined separation distance (gap length) where the increasing effective excitation numerical aperture is balanced by the decreasing confocal emission collection. A maximum signal is found at gap lengths of approximately 2, 1.25, and 1.75 mm for GRIN lenses with pitches of 0.23, 0.25, and 0.29 wavelength at 830 nm. The maximum two-photon fluorescence signal collected corresponds to a threefold reduction of axial resolution (38.5 µm at 1.25 mm), compared with the maximum resolution (11.6 µm at 5.5 mm), as shown by the three-dimensional imaging of 10 µm beads. These results demonstrate an intrinsic trade-off between signal collection and axial resolution.

  18. Linear and nonlinear optical properties of two-photon absorption dye doped linear copolymer

    Science.gov (United States)

    Wang, D.; Zhou, G. Y.; Ren, Y.; Yu, X. Q.; Cheng, X. F.; Yang, S. J.; Xu, X. G.; Shao, Z. S.; Jiang, M. H.

    2002-02-01

    2-hydroxyethyl methacrylate (abbreviated as HEMA) and acrylonitrile have been used as the monomers to form a trans-4-[p-(N-hydroxyethyl-N-methylamino)styryl]-N-methylpyridinium p-toluene sulfonate (abbreviated as HMASPS) doped linear copolymer. The spectra of one and two-photon excited fluorescence and two-photon pumped superradiance and two-photon pumped lasing of the polymer all shifted to short wavelengths compared with the solution sample of the dye. The copolymer shows much longer one- and two-photon excited fluorescence lifetimes of nanoseconds. When pumped by the picosecond Nd:YAG laser, the superradiance and lasing can be simultaneously obtained. The maximum two-photon pumped lasing conversion efficiency is 3.3%. The place of the maximum upconversion efficiency of the copolymer does not coincide with that of the maximum nonlinear absorption.

  19. Two-photon microscopy measurement of CMRO2 using periarteriolar PO2 gradients(Conference Presentation)

    Science.gov (United States)

    Sakadžić, Sava; Yaseen, Mohammad A.; Jaswal, Rajeshwer S.; Roussakis, Emmanuel; Dale, Anders M.; Buxton, Richard B.; Vinogradov, Sergei A.; Boas, David A.; Devor, Anna

    2017-02-01

    The cerebral metabolic rate of oxygen (CMRO2) is an essential parameter for evaluating brain function and pathophysiology. Measurements of CMRO2 with high spatio-temporal resolution are critically important for understanding how the brain copes with metabolic and blood perfusion changes associated with various clinical conditions, such as stroke, periinfarct depolarizations, and various microvasculopathies (e.g., Alzheimer's disease, chronic hypertension). CMRO2 measurements are also important for understanding the physiological underpinnings of functional Magnetic Resonance Imaging signals. However, the currently available approaches for quantifying CMRO2 rely on complex multimodal imaging and mathematical modeling. Here, we introduce a novel method that allows estimation of CMRO2 based on a single measurement modality - two-photon phosphorescence lifetime microscopy (2PLM) imaging of the partial pressure of oxygen (PO2) in cortical tissue. CMRO2 is estimated by fitting the changes of tissue PO2 around cortical penetrating arterioles with the Krogh cylinder model of oxygen diffusion. We measured the baseline CMRO2 in anesthetized rats, and modulated tissue PO2 levels by manipulating the depth of anesthesia. This method has a spatial resolution of approximately 200 μm and it may provide CMRO2 measurements in individual cortical layers or within confined cortical regions such as in ischemic penumbra and the foci of functional activation.

  20. Ultrafast axial scanning for two-photon microscopy via a digital micromirror device and binary holography.

    Science.gov (United States)

    Cheng, Jiyi; Gu, Chenglin; Zhang, Dapeng; Wang, Dien; Chen, Shih-Chi

    2016-04-01

    In this Letter, we present an ultrafast nonmechanical axial scanning method for two-photon excitation (TPE) microscopy based on binary holography using a digital micromirror device (DMD), achieving a scanning rate of 4.2 kHz, scanning range of ∼180  μm, and scanning resolution (minimum step size) of ∼270  nm. Axial scanning is achieved by projecting the femtosecond laser to a DMD programmed with binary holograms of spherical wavefronts of increasing/decreasing radii. To guide the scanner design, we have derived the parametric relationships between the DMD parameters (i.e., aperture and pixel size), and the axial scanning characteristics, including (1) maximum optical power, (2) minimum step size, and (3) scan range. To verify the results, the DMD scanner is integrated with a custom-built TPE microscope that operates at 60 frames per second. In the experiment, we scanned a pollen sample via both the DMD scanner and a precision z-stage. The results show the DMD scanner generates images of equal quality throughout the scanning range. The overall efficiency of the TPE system was measured to be ∼3%. With the high scanning rate, the DMD scanner may find important applications in random-access imaging or high-speed volumetric imaging that enables visualization of highly dynamic biological processes in 3D with submillisecond temporal resolution.

  1. Reduction of the pulse duration of the ultrafast laser pulses of the Two-Photon Laser Scanning Microscopy (2PLSM

    Directory of Open Access Journals (Sweden)

    Reshak Ali

    2008-07-01

    Full Text Available Abstract Background We provide an update of our two-photon laser scanning microscope by compressing or reducing the broadening of the pulse width of ultrafast laser pulses for dispersion precompensation, to enable the pulses to penetrate deeply inside the sample. Findings The broadening comes as the pulses pass through the optical elements. We enhanced and modified the quality and the sharpness of images by enhancing the resolution using special polarizer namely Glan Laser polarizer GL10. This polarizer consists of two prisms separated by air space. This air separation between the two prisms uses to delay the red wavelength when the light leaves the first prism to the air then to second prism. We note a considerable enhancing with using the GL polarizer, and we can see the details of the leaf structure in early stages when we trying to get focus through z-stacks of images in comparison to exactly the same measurements without using GL polarizer. Hence, with this modification we able to reduce the time of exposure the sample to the laser radiation thereby we will reduce the probability of photobleaching and phototoxicity. When the pulse width reduced, the average power of the laser pulses maintained at a constant level. Significant enhancement is found between the two kinds of images of the Two-Photon Excitation Fluorescence (TPEF. Conclusion In summary reduction the laser pulse width allowed to collect more diffraction orders which will used to form the images. The more diffraction orders the higher resolution images.

  2. Two-photon absorption and two-photon circular dichroism of L-tryptophan in the near to far UV region

    Science.gov (United States)

    Vesga, Yuly; Hernandez, Florencio E.

    2017-09-01

    Herein we report on the first measurements of the two-photon absorption (TPA) spectrum of L-tryptophan in DMSO solution in the near to far UV region and the two-photon circular dichroism (TPCD) signal corresponding to a transition at 200 nm. We demonstrate the application of the Double L-scan technique in the near to far UV region to perform polarization dependent TPA measurements of chiral molecules. TPCD measurements below 400 nm reveal that chiral molecules in solution, such as tryptophan/DMSO, can undergo photochemical reactions in front of prolonged exposure to UV radiation.

  3. Sensitive and rapid detection of endogenous hydrogen sulfide distributing in different mouse viscera via a two-photon fluorescent probe.

    Science.gov (United States)

    Chen, Qian; Yang, Jinfeng; Li, Yinhui; Zheng, Jing; Yang, Ronghua

    2015-10-08

    Development of efficient methods for detection of endogenous H2S in living cells and tissues is of considerable significance for better understanding the biological and pathological functions of H2S. Two-photon (TP) fluorescent probes are favorable as powerful molecular tools for studying physiological process due to its non-invasiveness, high spatiotemporal resolution and deep-tissues imaging. Up to date, several TP probes for intracellular H2S imaging have been designed, but real-time imaging of endogenous H2S-related biological processes in tissues is hampered due to low sensitivity, long response time and interference from other biothiols. To address this issue, we herein report a novel two-photon fluorescent probe (TPP-H2S) for highly sensitive and fast monitoring and imaging H2S levels in living cells and tissues. In the presence of H2S, it exhibits obviously improved sensitivity (LOD: 0.12 μM) and fast response time (about 2 min) compared with the reported two-photon H2S probes. With two-photon excitation, TPP-H2S displays high signal-to-noise ratio and sensitivity even no interference in cell growth media. As further application, TPP-H2S is applied for fast imaging of H2S in living cells and different fresh tissues by two-photon confocal microscope. Most importantly we first measured the endogenous H2S level in different viscera by vivisection and found that the distribution of endogenous H2S mostly in brain, liver and lung. The excellent sensing properties of TPP-H2S make it a practically useful tool for further studying biological roles of H2S. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Facile synthesis of two-photon absorbing polymers through radical copolymerization

    Directory of Open Access Journals (Sweden)

    2007-08-01

    Full Text Available A two-photon absorbing polymer has been prepared through radical copolymerization of methyl acrylate and a synthesized monomer containing a two-photon absorbing chromophore (E,E,E-1,3,5-tristyrylbenzene (1, under conventional radical polymerization conditions. The synthesized polymer was characterized by nuclear magnetic resonance (NMR, infra-red spectroscopy (IR and gel permeation chromatography (GPC. The linear and nonlinear optical properties were studied by measurement of UV-Vis absorption, fluorescent emission and two-photon cross-section. This synthetic strategy provided a facile approach for synthesis of photonic materials with adjustable chromophore concentration and high molecular weights.

  5. The study of nonlinear two-photon phenomenon in photonic crystals doped with nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Mahi R [Department of Physics and Astronomy, University of Western Ontario, London, N6A 3K7 (Canada)

    2007-02-28

    A theory of the nonlinear two-photon absorption has been developed in a photonic crystal doped with an ensemble of four-level nanoparticles. We have considered that the nanoparticles are interacting with the photonic crystal. An expression of two-photon absorption has been obtained by using the density matrix method. The effect of the dipole-dipole interaction has also been included in the formulation. Interesting new phenomena have been predicted. For example, it is found that the inhibition of two-photon absorption can be turned on and off when the decay resonance energies of the four-level nanoparticles are moved within the energy band.

  6. The two-photon exchange contribution to elastic electron-nucleon scattering at large momentum transfer

    Energy Technology Data Exchange (ETDEWEB)

    Andrei V. Afanasev; Stanley J. Brodsky; Carl E. Carlson; Yu-Chun Chen; Marc Vanderhaeghen

    2005-01-01

    We estimate the two-photon exchange contribution to elastic electron-proton scattering at large momentum transfer by using a quark-parton representation of virtual Compton scattering. We thus can relate the two-photon exchange amplitude to the generalized parton distributions which also enter in other wide angle scattering processes. We find that the interference of one- and two-photon exchange contribution is able to substantially resolve the difference between electric form factor measurements from Rosenbluth and polarization transfer experiments.

  7. Toward two-photon excited fluorescence lifetime endomicroscopy (Conference Presentation)

    Science.gov (United States)

    Hage, Charles-Henri; Leclerc, Pierre; Fabert, Marc; Brevier, Julien; Habert, Rémi; Braud, Flavie; Kudlinski, Alexandre; Louradour, Frédéric

    2017-02-01

    Fluorescence lifetime imaging microscopy (FLIM) represents a powerful tool for biological studies. Endoscopic FLIM applied to the intracellular native biomarker NADH and FAD represents a promising mean for in vivo in situ malignant tissue diagnosis in the medical field. Else, 2-photon-excited fluorescence (2PEF) provides increased 3D resolution and imaging depth. But very few demonstrations about 2PEF lifetime measurement through a fiber have been reported and none about endoscopic 2P-FLIM through a practical fiber length (FLIM. Our goal is now to check that collecting fluorescence through the same endoscopic fiber does not deteriorate the lifetime measurement. Relying on the basis previously published in case of 1PEF by P. French and co-workers (J. Biophotonics, 2015), we have experimentally quantitatively evaluated the influence on the lifetime measurement of the fiber chromatic and intermodal dispersions. The main result is that the fiber contribution to the system impulse response function, even in the case of a 3-meter long double-clad optical fiber, does not hinder the separation between free and bound NADH states using FLIM. Related calibrations and measurements will be detailed. Ongoing experiments about the development of a 2P-FLIM endomicroscope on the basis of an previously reported 2P-endomicroscope (Ducourthial et al., Sc. Reports, 2015), used under various configurations (i.e. point measurement in the center of the 2P-endomicroscope image, averaged lifetime, binned endoscopic 2P-FLIM image), will be also presented.

  8. Deep in vivo two-photon microscopy with a low cost custom built mode-locked 1060 nm fiber laser.

    Science.gov (United States)

    Perillo, Evan P; McCracken, Justin E; Fernée, Daniel C; Goldak, John R; Medina, Flor A; Miller, David R; Yeh, Hsin-Chih; Dunn, Andrew K

    2016-02-01

    Here we demonstrate that a mode-locked ytterbium fiber laser for two-photon fluorescence microscopy can be built for $13,000. The laser emits at a wavelength of 1060 nm with a usable average power of 1 W at a repetition rate of 40 MHz and a compressed pulse width of 81 fs at the sample. The laser is used to obtain deep in vivo two-color images of layer-V pyramidal neurons expressing YFP and vasculature labelled with Texas Red at depths up to 900 µm. The sub-1 µm features of dendritic spines can be resolved at a 200 µm depth.

  9. 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.

  10. Dynamic characterization of hydrophobic and hydrophilic solutes in oleic-acid enhanced transdermal delivery using two-photon fluorescence microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Tseng, Te-Yu; Yang, Chiu-Sheng; Chen, Yang-Fang [Department of Physics, National Taiwan University, Taipei, Taiwan (China); Tsai, Tsung-Hua [Department of Dermatology, Far Eastern Memorial Hospital, New Taipei City, Taiwan (China); Dong, Chen-Yuan, E-mail: cydong@phys.ntu.edu.tw [Department of Physics, National Taiwan University, Taipei, Taiwan (China); Center for Quantum Science and Engineering, National Taiwan University, Taipei, Taiwan (China); Center for Optoelectronic Biomedicine, National Taiwan University, Taipei, Taiwan (China)

    2014-10-20

    In this letter, we propose an efficient methodology of investigating dynamic properties of sulforhodamine B and rhodamine B hexyl ester molecules transporting across ex-vivo human stratum corneum with and without oleic acid enhancement. Three-dimensional, time-lapse fluorescence images of the stratum corneum can be obtained using two-photon fluorescence microscopy. Furthermore, temporal quantifications of transport enhancements in diffusion parameters can be achieved with the use of Fick's second law. Dynamic characterization of solutes transporting across the stratum corneum is an effective method for understanding transient phenomena in transdermal delivery of probe molecules, leading to improved delivery strategies of molecular species for therapeutic purposes.

  11. 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...

  12. Active stabilization of a fiber-optic two-photon interferometer using continuous optical length control

    National Research Council Canada - National Science Library

    Cho, Seok-Beom; Kim, Heonoh

    2016-01-01

    ... 6-km-long fiber-optic Hong-Ou-Mandel interferometer. The two-step active control techniques are applied for measuring highly stable two-photon interference fringes by scanning the optical path-length difference...

  13. Limitations of two-level emitters as nonlinearities in two-photon controlled-phase gates

    Science.gov (United States)

    Nysteen, Anders; McCutcheon, Dara P. S.; Heuck, Mikkel; Mørk, Jesper; Englund, Dirk R.

    2017-06-01

    We investigate the origin of imperfections in the fidelity of a two-photon controlled-phase gate based on two-level-emitter nonlinearities. We focus on a passive system that operates without external modulations to enhance its performance. We demonstrate that the fidelity of the gate is limited by opposing requirements on the input pulse width for one- and two-photon-scattering events. For one-photon scattering, the spectral pulse width must be narrow compared with the emitter linewidth, while two-photon-scattering processes require the pulse width and emitter linewidth to be comparable. We find that these opposing requirements limit the maximum fidelity of the two-photon controlled-phase gate to 84 % for photons with Gaussian spectral profiles.

  14. Hanbury Brown-Twiss effect without two-photon interference in photon counting regime.

    Science.gov (United States)

    Bai, Bin; Zhou, Yu; Liu, Ruifeng; Zheng, Huaibin; Wang, Yunlong; Li, Fuli; Xu, Zhuo

    2017-05-19

    From quantum point of view, Hanbury Brown-Twiss effect is a result of constructive-destructive two-photon interference. There should be no Hanbury Brown-Twiss effect if there was no two-photon interference. In this paper, we observed Hanbury Brown- Twiss effect in a specially designed experiment, in which two-photon interference is impossible by keeping only one two-photon probability amplitude in the experimental scheme. However, our experimental results can still be interpreted by Glauber's quantum optical coherence theory. The researches in our paper are helpful to understand the physics of the second-order coherence of light, especially the physics of Hanbury Brown-Twiss effect.

  15. A compact two photon light sheet microscope for applications in neuroscience

    DEFF Research Database (Denmark)

    Piksarv, Peeter; Marti, Dominik; Le, Tuan

    2016-01-01

    We present a compact setup for two photon light sheet microscopy. By using pulsed Airy beam illumination we demonstrate eight-fold increase of the FOV compared to Gaussian light sheet with the same axial resolution....

  16. Two-photon reactions with KLOE detector at DA$\\phi$NE

    CERN Document Server

    Ong, S

    1999-01-01

    We reexamine the feasibility of two-photon reactions at DA$\\Phi$NE with the KLOE detector excluding the small angle tagging system. Event-rate predictions of interesting channels : $\\gamma \\gamma \\to \\pi^0, \\eta $ and $\\gamma \\gamma

  17. Mitochondria-targeted cationic porphyrin-triphenylamine hybrids for enhanced two-photon photodynamic therapy.

    Science.gov (United States)

    Hammerer, Fabien; Poyer, Florent; Fourmois, Laura; Chen, Su; Garcia, Guillaume; Teulade-Fichou, Marie-Paule; Maillard, Philippe; Mahuteau-Betzer, Florence

    2018-01-01

    The proof of concept for two-photon activated photodynamic therapy has already been achieved for cancer treatment but the efficiency of this approach still heavily relies on the availability of photosensitizers combining high two-photon absorption and biocompatibility. In this line we recently reported on a series of porphyrin-triphenylamine hybrids which exhibit high singlet oxygen production quantum yield as well as high two-photon absorption cross-sections but with a very poor cellular internalization. We present herein new photosensitizers of the same porphyrin-triphenylamine hybrid series but bearing cationic charges which led to strongly enhanced water solubility and thus cellular penetration. In addition the new compounds have been found localized in mitochondria that are preferential target organelles for photodynamic therapy. Altogether the strongly improved properties of the new series combined with their specific mitochondrial localization lead to a significantly enhanced two-photon activated photodynamic therapy efficiency. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Improved sensitivity for two-photon frequency-domain lifetime measurement.

    Science.gov (United States)

    Estrada, Arnold D; Dunn, Andrew K

    2010-06-21

    We demonstrate a method to improve the measurement sensitivity of two-photon frequency-domain lifetime measurements in poor signal to background conditions. This technique uses sinusoidal modulation of the two-photon excitation source and detection of the second harmonic of the modulation frequency that appears in the emission. Additionally, we present the mathematical model which describes how the observed phase shift and amplitude demodulation factor of two-photon phosphorescence emission are related to the phosphorescence lifetime and modulation frequency. We demonstrate the validity of the model by showing the existence of new frequency terms in the phosphorescence emission generated from the quadratic nature of two-photon absorption and by showing that the phase shift and demodulation match theory for all frequency components.

  19. 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.

  20. Influence of Two Photon Absorption on Soliton Self-Frequency Shift

    DEFF Research Database (Denmark)

    Steffensen, Henrik; Rottwitt, Karsten; Jepsen, Peter Uhd

    2011-01-01

    The creation of mid-infrared supercontinua necessitates the use of soft-glass fibers. However, some materials, like chalcogenide, have a substantial two photon absorption. We introduce a model for soliton self-frequency shift that successfully includes this effect.......The creation of mid-infrared supercontinua necessitates the use of soft-glass fibers. However, some materials, like chalcogenide, have a substantial two photon absorption. We introduce a model for soliton self-frequency shift that successfully includes this effect....

  1. 3D two-photon lithographic microfabrication system

    Science.gov (United States)

    Kim, Daekeun [Cambridge, MA; So, Peter T. C. [Boston, MA

    2011-03-08

    An imaging system is provided that includes a optical pulse generator for providing an optical pulse having a spectral bandwidth and includes monochromatic waves having different wavelengths. A dispersive element receives a second optical pulse associated with the optical pulse and disperses the second optical pulse at different angles on the surface of the dispersive element depending on wavelength. One or more focal elements receives the dispersed second optical pulse produced on the dispersive element. The one or more focal element recombine the dispersed second optical pulse at a focal plane on a specimen where the width of the optical pulse is restored at the focal plane.

  2. Visualization of laser tattoo removal treatment effects in a mouse model by two-photon microscopy

    Science.gov (United States)

    Jang, Won Hyuk; Yoon, Yeoreum; Kim, Wonjoong; Kwon, Soonjae; Lee, Seunghun; Song, Duke; Choi, Jong Woon; Kim, Ki Hean

    2017-01-01

    Laser tattoo removal is an effective method of eliminating tattoo particles in the skin. However, laser treatment cannot always remove the unwanted tattoo completely, and there are risks of either temporary or permanent side effects. Studies using preclinical animal models could provide detailed information on the effects of laser treatment in the skin, and might help to minimize side effects in clinical practices. In this study, two-photon microscopy (TPM) was used to visualize the laser treatment effects on tattoo particles in both phantom specimens and in vivo mouse models. Fluorescent tattoo ink was used for particle visualization by TPM, and nanosecond (ns) and picosecond (ps) lasers at 532 nm were used for treatment. In phantom specimens, TPM characterized the fragmentation of individual tattoo particles by tracking them before and after the laser treatment. These changes were confirmed by field emission scanning electron microscopy (FE-SEM). TPM was used to measure the treatment efficiency of the two lasers at different laser fluences. In the mouse model, TPM visualized clusters of tattoo particles in the skin and detected their fragmentation after the laser treatment. Longitudinal TPM imaging observed the migration of cells containing tattoo particles after the laser treatment. These results show that TPM may be useful for the assessment of laser tattoo removal treatment in preclinical studies. PMID:28856046

  3. Visualization of laser tattoo removal treatment effects in a mouse model by two-photon microscopy.

    Science.gov (United States)

    Jang, Won Hyuk; Yoon, Yeoreum; Kim, Wonjoong; Kwon, Soonjae; Lee, Seunghun; Song, Duke; Choi, Jong Woon; Kim, Ki Hean

    2017-08-01

    Laser tattoo removal is an effective method of eliminating tattoo particles in the skin. However, laser treatment cannot always remove the unwanted tattoo completely, and there are risks of either temporary or permanent side effects. Studies using preclinical animal models could provide detailed information on the effects of laser treatment in the skin, and might help to minimize side effects in clinical practices. In this study, two-photon microscopy (TPM) was used to visualize the laser treatment effects on tattoo particles in both phantom specimens and in vivo mouse models. Fluorescent tattoo ink was used for particle visualization by TPM, and nanosecond (ns) and picosecond (ps) lasers at 532 nm were used for treatment. In phantom specimens, TPM characterized the fragmentation of individual tattoo particles by tracking them before and after the laser treatment. These changes were confirmed by field emission scanning electron microscopy (FE-SEM). TPM was used to measure the treatment efficiency of the two lasers at different laser fluences. In the mouse model, TPM visualized clusters of tattoo particles in the skin and detected their fragmentation after the laser treatment. Longitudinal TPM imaging observed the migration of cells containing tattoo particles after the laser treatment. These results show that TPM may be useful for the assessment of laser tattoo removal treatment in preclinical studies.

  4. 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.

  5. Two-photon microscopy for non-invasive, quantitative monitoring of stem cell differentiation.

    Directory of Open Access Journals (Sweden)

    William L Rice

    Full Text Available BACKGROUND: The engineering of functional tissues is a complex multi-stage process, the success of which depends on the careful control of culture conditions and ultimately tissue maturation. To enable the efficient optimization of tissue development protocols, techniques suitable for monitoring the effects of added stimuli and induced tissue changes are needed. METHODOLOGY/PRINCIPAL FINDINGS: Here, we present the quantitative use of two-photon excited fluorescence (TPEF and second harmonic generation (SHG as a noninvasive means to monitor the differentiation of human mesenchymal stem cells (hMSCs using entirely endogenous sources of contrast. We demonstrate that the individual fluorescence contribution from the intrinsic cellular fluorophores NAD(PH, flavoproteins and lipofuscin can be extracted from TPEF images and monitored dynamically from the same cell population over time. Using the redox ratio, calculated from the contributions of NAD(PH and flavoproteins, we identify distinct patterns in the evolution of the metabolic activity of hMSCs maintained in either propagation, osteogenic or adipogenic differentiation media. The differentiation of these cells is mirrored by changes in cell morphology apparent in high resolution TPEF images and by the detection of collagen production via SHG imaging. Finally, we find dramatic increases in lipofuscin levels in hMSCs maintained at 20% oxygen vs. those in 5% oxygen, establishing the use of this chromophore as a potential biomarker for oxidative stress. CONCLUSIONS/SIGNIFICANCE: In this study we demonstrate that it is possible to monitor the metabolic activity, morphology, ECM production and oxidative stress of hMSCs in a non-invasive manner. This is accomplished using generally available multiphoton microscopy equipment and simple data analysis techniques, such that the method can widely adopted by laboratories with a diversity of comparable equipment. This method therefore represents a powerful tool

  6. Dynamic performance of microelectromechanical systems deformable mirrors for use in an active/adaptive two-photon microscope

    Science.gov (United States)

    Archer-Zhang, Christian Chunzi; Foster, Warren B.; Downey, Ryan D.; Arrasmith, Christopher L.; Dickensheets, David L.

    2016-12-01

    Active optics such as deformable mirrors can be used to control both focal depth and aberrations during scanning laser microscopy. If the focal depth can be changed dynamically during scanning, then imaging of oblique surfaces becomes possible. If aberrations can be corrected dynamically during scanning, an image can be optimized throughout the field of view. Here, we characterize the speed and dynamic precision of a Boston Micromachines Corporation Multi-DM 140 element aberration correction mirror and a Revibro Optics 4-zone focus control mirror to assess suitability for use in an active and adaptive two-photon microscope. Tests for the multi-DM include both step response and sinusoidal frequency sweeps of specific Zernike modes (defocus, spherical aberration, coma, astigmatism, and trefoil). We find wavefront error settling times for mode amplitude steps as large as 400 nm to be less than 52 μs, with 3 dB frequencies ranging from 6.5 to 10 kHz. The Revibro Optics mirror was tested for step response only, with wavefront error settling time less than 80 μs for defocus steps up to 3000 nm, and less than 45 μs for spherical aberration steps up to 600 nm. These response speeds are sufficient for intrascan correction at scan rates typical of two-photon microscopy.

  7. Multi-point scanning two-photon excitation microscopy by utilizing a high-peak-power 1042-nm laser.

    Science.gov (United States)

    Otomo, Kohei; Hibi, Terumasa; Murata, Takashi; Watanabe, Hirotaka; Kawakami, Ryosuke; Nakayama, Hiroshi; Hasebe, Mitsuyasu; Nemoto, Tomomi

    2015-01-01

    The temporal resolution of a two-photon excitation laser scanning microscopy (TPLSM) system is limited by the excitation laser beam's scanning speed. To improve the temporal resolution, the TPLSM system is equipped with a spinning-disk confocal scanning unit. However, the insufficient energy of a conventional Ti:sapphire laser source restricts the field of view (FOV) for TPLSM images to a narrow region. Therefore, we introduced a high-peak-power Yb-based laser in order to enlarge the FOV. This system provided three-dimensional imaging of a sufficiently deep and wide region of fixed mouse brain slices, clear four-dimensional imaging of actin dynamics in live mammalian cells and microtubule dynamics during mitosis and cytokinesis in live plant cells.

  8. 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.

  9. Two-photon Photoemission of Organic Semiconductor Molecules on Ag(111)

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Aram [Univ. of California, Berkeley, CA (United States)

    2008-05-01

    Angle- and time-resolved two-photon photoemission (2PPE) was used to study systems of organic semiconductors on Ag(111). The 2PPE studies focused on electronic behavior specific to interfaces and ultrathin films. Electron time dynamics and band dispersions were characterized for ultrathin films of a prototypical n-type planar aromatic hydrocarbon, PTCDA, and representatives from a family of p-type oligothiophenes.In PTCDA, electronic behavior was correlated with film morphology and growth modes. Within a fewmonolayers of the interface, image potential states and a LUMO+1 state were detected. The degree to which the LUMO+1 state exhibited a band mass less than a free electron mass depended on the crystallinity of the layer. Similarly, image potential states were measured to have free electron-like effective masses on ordered surfaces, and the effective masses increased with disorder within the thin film. Electron lifetimes were correlated with film growth modes, such that the lifetimes of electrons excited into systems created by layer-by-layer, amorphous film growth increased by orders of magnitude by only a few monolayers from the surface. Conversely, the decay dynamics of electrons in Stranski-Krastanov systems were limited by interaction with the exposed wetting layer, which limited the barrier to decay back into the metal.Oligothiophenes including monothiophene, quaterthiophene, and sexithiophene were deposited on Ag(111), and their electronic energy levels and effective masses were studied as a function of oligothiophene length. The energy gap between HOMO and LUMO decreased with increasing chain length, but effective mass was found to depend on domains from high- or low-temperature growth conditions rather than chain length. In addition, the geometry of the molecule on the surface, e.g., tilted or planar, substantially affected the electronic structure.

  10. Quantifying the cellular uptake of antibody-conjugated Au nanocages by two-photon microscopy and inductively coupled plasma mass spectrometry.

    Science.gov (United States)

    Au, Leslie; Zhang, Qiang; Cobley, Claire M; Gidding, Michael; Schwartz, Andrea G; Chen, Jingyi; Xia, Younan

    2010-01-26

    Gold nanocages with localized surface plasmon resonance peaks in the near-infrared region exhibited a broad two-photon photoluminescence band extending from 450 to 650 nm when excited by a Ti:sapphire laser at 800 nm. The bright luminescence makes it possible to explore the use of Au nanocages as a new class of optical imaging agents for two-photon microscopy. In this work, we have demonstrated the use of two-photon microscopy as a convenient tool to directly examine the uptake of antibody-conjugated and PEGylated Au nanocages by U87MGwtEGFR cells. We have also correlated the results from two-photon microscopy with the data obtained by inductively coupled plasma mass spectrometry. Combined together, these results indicate that the antibody-conjugated Au nanocages were attached to the surface of the cells through antibody-antigen binding and then internalized into the cells via receptor-mediated endocytosis. The cellular uptake process was dependent on a number of parameters, including incubation time, incubation temperature, size of the Au nanocages, and the number of antibodies immobilized on each nanocage.

  11. Metal Complexes for Two-Photon Photodynamic Therapy: A Cyclometallated Iridium Complex Induces Two-Photon Photosensitization of Cancer Cells under Near-IR Light

    OpenAIRE

    Mckenzie, L.; Sazanovich, I.; Baggaley, E.; Bonneau, M.; Guerchais, V.; Williams, G; Weinstein, J; Bryant, H. E.

    2017-01-01

    International audience; Photodynamic therapy (PDT) uses photosensitizers (PS) which only become cytotoxic upon light-irradiation. Transition-metal complexes are highly promising PS due to long excited-state lifetimes, and high photo-stabilities. However, these complexes usually absorb higher-energy UV/Vis light, whereas the optimal tissue transparency is in the lower-energy NIR region. Two-photon excitation (TPE) can overcome this dichotomy, with simultaneous absorption of two lower-energy NI...

  12. Note: Derivation of two-photon circular dichroism - Addendum to "two-photon circular dichroism" [J. Chem. Phys. 62, 1006 (1975)

    OpenAIRE

    Friese, Daniel Henrik

    2015-01-01

    Published version, also available at http://dx.doi.org/10.1063/1.4930017 This addendum shows the detailed derivation of the fundamental equations for two-photon circular dichroism which are given in a very condensed form in the original publication [I. Tinoco, J. Chem. Phys. 62, 1006 (1975)]. In addition, some minor errors are corrected and some of the derivations in the original publication are commented.

  13. Two-photon photosensitized production of singlet oxygen: optical and optoacoustic characterization of absolute two-photon absorption cross sections for standard sensitizers in different solvents.

    Science.gov (United States)

    Arnbjerg, Jacob; Johnsen, Mette; Frederiksen, Peter K; Braslavsky, Silvia E; Ogilby, Peter R

    2006-06-15

    Singlet molecular oxygen, O2(a1Deltag), can be produced upon resonant two-photon excitation of a photosensitizer. In the present study, two molecules that have received recent attention in studies of nonlinear organic materials were characterized for use as standard two-photon sensitizers: 2,5-dicyano-1,4-bis(2-(4-diphenylaminophenyl)vinyl)-benzene, CNPhVB, and 2,5-dibromo-1,4-bis(2-(4-diphenylaminophenyl)vinyl)-benzene, BrPhVB. Absolute two-photon absorption cross sections, delta, were independently determined for these molecules using two techniques that have heretofore not been applied to this problem: an optical technique (time-resolved detection of O2(a1Deltag) phosphorescence) and a nonoptical technique (a time-resolved laser-induced optoacoustic experiment). For experiments performed in toluene, a solvent commonly used for such nonlinear optical studies, appreciable absorption by the solvent itself complicates the measurements. In cyclohexane, however, delta values could be obtained without the interfering effects of solvent absorption. On the basis of these results, we discuss key aspects of the respective techniques used to quantify values of delta. The information reported herein provides some explanation for the lack of consensus that is routinely observed in published values of delta, certainly for experiments performed in aromatic solvents such as toluene and benzene.

  14. Pseudopotential calculations and photothermal lensing measurements of two-photon absorption in solids

    Energy Technology Data Exchange (ETDEWEB)

    White, W.T. III

    1985-11-04

    We have studied two-photon absorption in solids theoretically and experimentally. We have shown that it is possible to use accurate band structure techniques to compute two-photon absorption spectra within 15% of measured values in a wide band-gap material, ZnS. The empirical pseudopotential technique that we used is significantly more accurate than previous models of two-photon absorption in zinc blende materials, including present tunneling theories (which are essentially parabolic-band results in disguise) and the nonparabolic-band formalism of Pidgeon et al. and Weiler. The agreement between our predictions and previous measurements allowed us to use ZnS as a reference material in order to validate a technique for measuring two-photon absorption that was previously untried in solids, pulsed dual-beam thermal lensing. With the validated technique, we examined nonlinear absorption in one other crystal (rutile) and in several glasses, including silicates, borosilicates, and one phosphate glass. Initially, we believed that the absorption edges of all the materials were comparable; however, subsequent evidence suggested that the effective band-gap energies of the glasses were above the energy of two photons in our measurement. Therefore, we attribute the nonlinear absorption that we observed in glasses to impurities or defects. The measured nonlinear absorption coefficients were of the order of a few cm/TW in the glasses and of the order of 10 cm/GW in the crystals, four orders of magnitude higher than in glasses. 292 refs.

  15. Optical Biomedical Diagnostics: Sensors with Optical Response Based on Two-Photon Excited Luminescent Dyes for Biomolecules Detection

    Directory of Open Access Journals (Sweden)

    V. M. Yashchuk

    2008-01-01

    Full Text Available The spectral properties of novel styryl dyes developed for the biomacromolecules (such as DNA detection and imaging were investigated. The energy structures of dye molecules were examined. The spectral data prove that dyes aggregate and interact with DNA. The essential increase of the fluorescence intensity of dyes in the presence of DNA was observed. The photostability and phototoxic influence on the DNA of several styryl dyes were studied by analyzing absorption, fluorescence, and phosphorescence spectra of these dyes and dye-DNA systems. Changes of the optical density value of dye-DNA solutions caused by the irradiation were fixed in the DNA and dye absorption wavelength regions. Fluorescence emission of dye-DNA complexes upon two-photon excitation at wavelength 1064 nm with the 20-nanosecond pulsed YAG:Nd3+ laser and at 840 nm with the 90 famtosecond pulsed Ti:sapphire laser was registered. The values of two-photon absorption cross-sections of dye-DNA complexes were evaluated.

  16. Active stabilization of a fiber-optic two-photon interferometer using continuous optical length control.

    Science.gov (United States)

    Cho, Seok-Beom; Kim, Heonoh

    2016-05-16

    The practical realization of long-distance entanglement-based quantum communication systems strongly rely on the observation of highly stable quantum interference between correlated single photons. This task must accompany active stabilization of the optical path lengths within the single-photon coherence length. Here, we provide two-step interferometer stabilization methods employing continuous optical length control and experimentally demonstrate two-photon quantum interference using an actively stabilized 6-km-long fiber-optic Hong-Ou-Mandel interferometer. The two-step active control techniques are applied for measuring highly stable two-photon interference fringes by scanning the optical path-length difference. The obtained two-photon interference visibilities with and without accidental subtraction are found to be approximately 90.7% and 65.4%, respectively.

  17. 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.

  18. Fabrication of microneedles using two photon polymerization for transdermal delivery of nanomaterials.

    Science.gov (United States)

    Doraiswamy, Anand; Ovsianikov, Aleksandr; Gittard, Shaun D; Monteiro-Riviere, Nancy A; Crombez, Rene; Montalvo, Eva; Shen, Weidian; Chichkov, Boris N; Narayan, Roger J

    2010-10-01

    Microneedle devices for transdermal delivery of nanoscale pharmacologic agents were fabricated out of organically-modified ceramic (Ormocer) materials using two photon polymerization. Out-of-plane hollow microneedle arrays with various aspect ratios were fabricated using this rapid prototyping process. Human epidermal keratinocyte (HEK) viability on Ormocer surfaces fabricated using two photon polymerization was similar to that on control surfaces. Nanoindentation studies were performed to determine hardness and Young's modulus values for Ormocer materials. Microneedies were shown to enable more rapid distribution of the PEG-amine quantum dot solution to the deep epidermis and dermis layers of porcine skin than topical administration. Our results suggest that two photon polymerization may be used to create microneedle arrays for transdermal delivery of nanoscale pharmacologic agents.

  19. THz/Infrared Double Resonance Two-Photon Spectroscopy of HD+ for Determination of Fundamental Constants

    Directory of Open Access Journals (Sweden)

    Florin Lucian Constantin

    2017-10-01

    Full Text Available A double resonance two-photon spectroscopy scheme is discussed to probe jointly rotational and rovibrational transitions of ensembles of trapped HD+ ions. The two-photon transition rates and lightshifts are calculated with the two-photon tensor operator formalism. The rotational lines may be observed with sub-Doppler linewidth at the hertz level and good signal-to-noise ratio, improving the resolution in HD+ spectroscopy beyond the 10−12 level. The experimental accuracy, estimated at the 10−12 level, is comparable with the accuracy of theoretical calculations of HD+ energy levels. An adjustment of selected rotational and rovibrational HD+ lines may add clues to the proton radius puzzle, may provide an independent determination of the Rydberg constant, and may improve the values of proton-to-electron and deuteron-to-proton mass ratios beyond the 10−11 level.

  20. 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.

  1. Resonance enhancement of two photon absorption by magnetically trapped atoms in strong rf-fields

    Science.gov (United States)

    Chakraborty, A.; Mishra, S. R.

    2018-01-01

    Applying a many mode Floquet formalism for magnetically trapped atoms interacting with a polychromatic rf-field, we predict a large two photon transition probability in the atomic system of cold 87Rb atoms. The physical origin of this enormous increase in the two photon transition probability is due to the formation of avoided crossings between eigen-energy levels originating from different Floquet sub-manifolds and redistribution of population in the resonant intermediate levels to give rise to the resonance enhancement effect. Other exquisite features of the studied atom-field composite system include the splitting of the generated avoided crossings at the strong field strength limit and a periodic variation of the single and two photon transition probabilities with the mode separation frequency of the polychromatic rf-field. This work can find applications to characterize properties of cold atom clouds in the magnetic traps using rf-spectroscopy techniques.

  2. 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.

  3. Search for a Higgs Boson Decaying into Two Photons at LEP

    CERN Document Server

    Achard, P.; Aguilar-Benitez, M.; Alcaraz, J.; Alemanni, G.; Allaby, J.; Aloisio, A.; Alviggi, M.G.; Anderhub, H.; Andreev, Valery 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.; Barillere, R.; Bartalini, P.; Basile, M.; Batalova, N.; Battiston, R.; Bay, A.; Becattini, F.; Becker, U.; Behner, F.; Bellucci, L.; Berbeco, R.; Berdugo, J.; Berges, P.; Bertucci, B.; Betev, B.L.; Biasini, M.; Biglietti, M.; Biland, A.; Blaising, J.J.; Blyth, S.C.; Bobbink, G.J.; Bohm, A.; Boldizsar, L.; Borgia, B.; Bottai, S.; Bourilkov, D.; Bourquin, M.; Braccini, S.; Branson, J.G.; Brochu, F.; Burger, J.D.; Burger, W.J.; Cai, X.D.; Capell, M.; Cara Romeo, G.; Carlino, G.; Cartacci, A.; Casaus, J.; Cavallari, F.; Cavallo, N.; Cecchi, C.; Cerrada, M.; Chamizo, M.; Chang, Y.H.; Chemarin, M.; Chen, A.; Chen, G.; Chen, G.M.; Chen, H.F.; Chen, H.S.; Chiefari, G.; Cifarelli, L.; Cindolo, F.; Clare, I.; Clare, R.; Coignet, G.; Colino, N.; Costantini, S.; de la Cruz, B.; Cucciarelli, S.; van Dalen, J.A.; de Asmundis, R.; Deglon, P.; Debreczeni, J.; Degre, A.; Deiters, K.; della Volpe, D.; Delmeire, E.; Denes, P.; DeNotaristefani, F.; De Salvo, A.; Diemoz, M.; Dierckxsens, M.; Dionisi, C.; Dittmar, M.; Doria, A.; Dova, M.T.; Duchesneau, D.; Echenard, B.; Eline, A.; El Mamouni, H.; Engler, A.; Eppling, F.J.; Ewers, A.; Extermann, P.; Falagan, M.A.; Falciano, S.; Favara, A.; Fay, J.; Fedin, O.; Felcini, M.; Ferguson, T.; Fesefeldt, H.; Fiandrini, E.; Field, J.H.; Filthaut, F.; Fisher, P.H.; Fisher, W.; Fisk, I.; Forconi, G.; Freudenreich, K.; Furetta, C.; Galaktionov, Iouri; Ganguli, S.N.; Garcia-Abia, Pablo; Gataullin, M.; Gentile, S.; Giagu, S.; Gong, Z.F.; Grenier, Gerald Jean; Grimm, O.; Gruenewald, M.W.; Guida, M.; van Gulik, R.; Gupta, V.K.; Gurtu, A.; Gutay, L.J.; Haas, D.; Hakobyan, R.S.; Hatzifotiadou, D.; Hebbeker, T.; Herve, Alain; Hirschfelder, J.; Hofer, H.; Hohlmann, M.; Holzner, G.; Hou, S.R.; Hu, Y.; Jin, B.N.; Jones, Lawrence W.; de Jong, P.; Josa-Mutuberria, I.; Kafer, D.; Kaur, M.; Kienzle-Focacci, M.N.; Kim, J.K.; Kirkby, Jasper; Kittel, W.; Klimentov, A.; Konig, A.C.; Kopal, M.; Koutsenko, V.; Kraber, M.; Kraemer, R.W.; Krenz, W.; Kruger, A.; Kunin, A.; Ladron 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.; Levtchenko, P.; Li, C.; Likhoded, S.; Lin, C.H.; Lin, W.T.; Linde, F.L.; Lista, L.; Liu, Z.A.; Lohmann, W.; Longo, E.; Lu, Y.S.; Lubelsmeyer, K.; Luci, C.; Luminari, L.; Lustermann, W.; Ma, W.G.; Malgeri, L.; Malinin, A.; Mana, C.; Mangeol, D.; 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.; Kluge, Hannelies; Ofierzynski, R.; Organtini, G.; Palomares, C.; Pandoulas, D.; Paolucci, P.; Paramatti, R.; Passaleva, G.; Patricelli, S.; Paul, Thomas Cantzon; Pauluzzi, M.; Paus, C.; Pauss, F.; Pedace, M.; Pensotti, S.; Perret-Gallix, D.; Petersen, B.; Piccolo, D.; Pierella, F.; Pioppi, M.; Piroue, P.A.; Pistolesi, E.; Plyaskin, V.; Pohl, M.; Pojidaev, V.; Pothier, J.; Prokofev, D.O.; Prokofiev, D.; Quartieri, J.; Rahal-Callot, G.; Rahaman, M.A.; Raics, P.; Raja, N.; Ramelli, R.; Rancoita, P.G.; Ranieri, R.; Raspereza, A.; Razis, P.; Ren, D.; Rescigno, M.; Reucroft, S.; Riemann, S.; Riles, Keith; Roe, B.P.; Romero, L.; Rosca, A.; Rosier-Lees, S.; Roth, Stefan; Rosenbleck, C.; Roux, B.; Rubio, J.A.; Ruggiero, G.; Rykaczewski, H.; Sakharov, A.; Saremi, S.; Sarkar, S.; Salicio, J.; Sanchez, E.; Sanders, M.P.; Schafer, C.; Schegelsky, V.; Schmidt-Kaerst, S.; Schmitz, D.; Schopper, H.; Schotanus, D.J.; Schwering, G.; Sciacca, C.; Servoli, L.; Shevchenko, S.; Shivarov, N.; Shoutko, V.; Shumilov, E.; Shvorob, A.; Siedenburg, T.; Son, D.; Spillantini, P.; Steuer, M.; Stickland, D.P.; Stoyanov, B.; Straessner, A.; Sudhakar, K.; Sultanov, G.; Sun, L.Z.; Sushkov, S.; Suter, H.; Swain, J.D.; Szillasi, Z.; Tang, X.W.; Tarjan, P.; Tauscher, L.; Taylor, L.; Tellili, B.; Teyssier, D.; Timmermans, Charles; Ting, Samuel C.C.; Ting, S.M.; Tonwar, S.C.; Toth, J.; Tully, C.; Tung, K.L.; Ulbricht, J.; Valente, E.; Van de Walle, R.T.; Veszpremi, V.; Vesztergombi, G.; Vetlitsky, I.; Vicinanza, D.; Viertel, G.; Villa, S.; Vivargent, M.; Vlachos, S.; Vodopianov, 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.; Zoller, M.

    2002-01-01

    A Higgs particle produced in association with a Z boson and decaying into two photons is searched for in the data collected by the L3 experiment at LEP. All possible decay modes of the Z boson are investigated. No signal is observed in 447.5 pb^-1 of data recorded at centre-of-mass energies up to 209 GeV. Limits on the branching fraction of the Higgs boson decay into two photons as a function of the Higgs mass are derived. A lower limit on the mass of a fermiophobic Higgs boson is set at 105.4 GeV at 95% confidence level.

  4. Two-photon quantum walks in an elliptical direct-write waveguide array

    Science.gov (United States)

    Owens, J. O.; Broome, M. A.; Biggerstaff, D. N.; Goggin, M. E.; Fedrizzi, A.; Linjordet, T.; Ams, M.; Marshall, G. D.; Twamley, J.; Withford, M. J.; White, A. G.

    2011-07-01

    Integrated optics provides an ideal testbed for the emulation of quantum systems via continuous-time quantum walks. Here, we study the evolution of two-photon states in an elliptic array of waveguides. We characterize the photonic chip via coherent light tomography and use the results to predict distinct differences between temporally indistinguishable and distinguishable two-photon inputs, which we then compare with experimental observations. This work highlights the feasibility of emulation of coherent quantum phenomena in three-dimensional waveguide structures.

  5. Two-photon quantum walks in an elliptical direct-write waveguide array

    Energy Technology Data Exchange (ETDEWEB)

    Owens, J O; Broome, M A; Biggerstaff, D N; Goggin, M E; Fedrizzi, A; White, A G [ARC Centre for Engineered Quantum Systems, ARC Centre for Quantum Computer and Communication Technology, School of Mathematics and Physics, University of Queensland, Brisbane, QLD 4072 (Australia); Linjordet, T; Twamley, J [ARC Centre for Engineered Quantum Systems, Department of Physics and Astronomy, Macquarie University, North Ryde, NSW 2109 (Australia); Ams, M; Marshall, G D; Withford, M J, E-mail: m.a.broome@googlemail.com [ARC Centre for Ultrahigh Bandwidth Devices for Optical Systems, Centre for Quantum Science and Technology, MQ Photonics Research Centre, Department of Physics and Astronomy, Macquarie University, North Ryde, NSW 2109 (Australia)

    2011-07-15

    Integrated optics provides an ideal testbed for the emulation of quantum systems via continuous-time quantum walks. Here, we study the evolution of two-photon states in an elliptic array of waveguides. We characterize the photonic chip via coherent light tomography and use the results to predict distinct differences between temporally indistinguishable and distinguishable two-photon inputs, which we then compare with experimental observations. This work highlights the feasibility of emulation of coherent quantum phenomena in three-dimensional waveguide structures.

  6. Superluminality effect for laser pulse propagation in medium containing nanorods under the two-photon luminescence

    Science.gov (United States)

    Trofimov, Vyacheslav A.; Lysak, Tatyana M.

    2016-10-01

    We investigate the possibility for superluminality effects at the femtosecond pulse propagation in the medium with nanorods, if second harmonic generation (SGH) and two-photon luminescence (TPL) take place. On basis of semiclassical approach, we derive a set of equations, which describe the atomic states and electric fields dynamics. We show the possibility of light acceleration and soliton formation, if nanorod aspect ratio changing occurs due to their reshaping because two photon absorption (TPA) of optical energy, and investigate TPL and SHG in the medium with nanorods under sequential one-photon absorptions.

  7. In situ electrical and thermal monitoring of printed electronics by two-photon mapping

    DEFF Research Database (Denmark)

    Pastorelli, Francesco; Accanto, Nicolo; Jørgensen, Mikkel

    2017-01-01

    Printed electronics is emerging as a new, large scale and cost effective technology that will be disruptive in fields such as energy harvesting, consumer electronics and medical sensors. The performance of printed electronic devices relies principally on the carrier mobility and molecular packing......-destructive and low-cost testing method is needed. In this study, we demonstrate that nonlinear optical microscopy is a promising technique to achieve this goal. Using ultrashort laser pulses we stimulate two-photon absorption in a roll coated polymer semiconductor and map the resulting two-photon induced...

  8. Multi-focus two-photon polymerization technique based on individually controlled phase modulation.

    Science.gov (United States)

    Obata, Kotaro; Koch, Jürgen; Hinze, Ulf; Chichkov, Boris N

    2010-08-02

    Multi-focus two-photon polymerization with a spatial light modulator is demonstrated. The spatial light modulator generates multi-focus spots via phase modulation technique controlled by a computer generated hologram (CGH) pattern. Each focus spot can be individually addressed in position and laser intensity. The multi-focus two-photon polymerization technique allows the fabrication of complex 2-D and 3-D structures both symmetric and asymmetric. Smooth sine curved polymerized lines with amplitude of 5 microm and a period of 200 microm were obtained by fast switching of the CGH patterns.

  9. 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.

  10. From one-photon to two-photon probes: "caged" compounds, actuators, and photoswitches.

    Science.gov (United States)

    Bort, Guillaume; Gallavardin, Thibault; Ogden, David; Dalko, Peter I

    2013-04-22

    Molecular systems that can be remotely controlled by light are gaining increasing importance in cell biology, physiology, and neurosciences because of the spatial and temporal precision that is achievable with laser microscopy. Two-photon excitation has significant advantages deep in biological tissues, but raises problems in the design of "smart" probes compatible with cell physiology. This Review discusses the chemical challenges in generating suitable two-photon probes. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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

    DEFF Research Database (Denmark)

    Pedersen, Liselotte Jauffred; Kyrsting, Anders Højbo; 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...... two-photon absorption likely. This is in accordance with the observation that a trapped quantum dot is only fluorescing 5-10 percent of the time. The results are important for realizing nano-scale quantum dot control and visualization and for correct interpretation of experiments using two...

  12. 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.

  13. All possible bipartite positive-operator-value measurements of two-photon polarization states

    Science.gov (United States)

    Ahnert, S. E.; Payne, M. C.

    2006-02-01

    Here we propose an implementation of all possible positive-operator-value measures (POVMs) of two-photon polarization states. POVMs are the most general class of quantum measurements. Our setup requires linear optics, Bell state measurements, and an entangled three-photon ancilla state, which can be prepared separately and in advance (or “off-line”). As an example we give the detailed settings for a simultaneous measurement of all four Bell states for an arbitrary two-photon polarization state, which is impossible with linear optics alone.

  14. 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......, an increase in the field of view over the use of a standard Gaussian beam by a factor of six is demonstrated. This implementation for light sheet microscopy opens up new possibilities across a wide range of biomedical applications, especially for the study of neuronal processes....

  15. Higgs decay into two photons from a 3HDM with flavor symmetry

    Energy Technology Data Exchange (ETDEWEB)

    Aranda, Alfredo, E-mail: fefo@ucol.mx [Facultad de Ciencias, CUICBAS, Universidad de Colima, Colima (Mexico); Dual C-P Institute of High Energy Physics (Mexico); Bonilla, Cesar, E-mail: rasec.cmbd@gmail.com [Facultad de Ciencias Físico–Matemáticas, Benemérita Universidad Autónoma de Puebla (Mexico); Anda, Francisco de, E-mail: franciscojosedea@gmail.com [Departamento de Fisica, CUCEI, Universidad de Guadalajara (Mexico); Delgado, Antonio, E-mail: antonio.delgado@nd.edu [Department of Physics, University of Notre Dame, Notre Dame, IN 46556 (United States); Hernández-Sánchez, Jaime, E-mail: jaimeh@ece.buap.mx [Dual C-P Institute of High Energy Physics (Mexico); Facultad de Ciencias de la Electrónica, Benemérita Universidad Autónoma de Puebla, Apdo. Postal 542, 72570 Puebla, Puebla (Mexico)

    2013-08-09

    In this short Letter we show that the excess of events in the decay of Higgs to two photons reported by ATLAS and CMS can be easily accommodated in a flavor renormalizable three Higgs doublet model (3HDM). The model is consistent with all fermion masses, mixing angles, and flavor changing neutral current constraints.

  16. Two-photon excitation of atoms by ultrashort electromagnetic pulses in a discrete spectrum

    CERN Document Server

    Astapenko, Valery

    2015-01-01

    The present work is dedicated to the theoretical analysis of two-photon excitation of atoms in a discrete energy spectrum by ultrashort electromagnetic pulses of femto- and subfemtosecond ranges of durations. As examples, excitation of hydrogen and sodium atoms from the ground state to excited states with a zero orbital moment is considered.

  17. Combining Two-Photon Polymerisation and Photoreduction to Enable the Manufacture of Metamaterials at the Nanosale

    Science.gov (United States)

    2016-01-13

    laser beam is conditioned through a series of optics, e.g. a λ/2 wave plate, polarizer and beam expander , before it is divided into an objective lens...221102. Vurth, L., P. Baldeck, et al. (2008). "Two-photon induced fabrication of gold microstructures in polystyrene sulfonate thin films using a

  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. Programmable two-photon quantum interference in 10^3 channels in opaque scattering media

    NARCIS (Netherlands)

    Wolterink, Tom; Uppu, Ravitej; Ctistis, Georgios; Vos, Willem L.; Boller, Klaus J.; Pinkse, Pepijn Willemszoon Harry

    2016-01-01

    We investigate two-photon quantum interference in an opaque scattering medium that intrinsically supports a large number of transmission channels. By adaptive spatial phase modulation of the incident wave fronts, the photons are directed at targeted speckle spots or output channels. From 10 3

  20. Stability and bandgaps of layered perovskites for one- and two-photon water splitting

    DEFF Research Database (Denmark)

    Castelli, Ivano Eligio; García Lastra, Juan Maria; Hüser, Falco

    2013-01-01

    in the Ruddlesden–Popper phase of the layered perovskite structure. Based on screening criteria for the stability, bandgaps and band edge positions, we suggest 20 new materials for the light harvesting photo-electrode of a one-photon water splitting device and 5 anode materials for a two-photon device with silicon...

  1. Two-photon luminescence and stimulated emission depletion with gold nanorods by a single wavelength

    Science.gov (United States)

    Chen, Jianling; You, Minghai; Peng, Yiru; Yang, Hongqin; Xie, Shusen

    2017-06-01

    Single-wavelength two photon STED technique simplifies common STED setup, and decreases the cost of the system. However, this technique limits the fluorescent dye to a few kinds. Because the dye should be excited and depleted by the same single wavelength laser. And the gold nanorods can be excited by near-infrared pulsed laser and produces two photon luminescence, which is brighter than general two-photon autofluorescence by three orders. We found that the gold nanorods can also be depleted by near-infrared pulsed laser. We studied the stimulated emission depletion of gold nanorods with a single wavelength laser in the near infrared. Two photon luminescence was excited with a femtosecond pulse, then depleted by stimulated emission with a stretched pulse. The depletion efficiency with different wavelength was measured to pick out the optimal wavelength. We showed that this method can be applied to super-resolved STED microscopy, which combines the high brightness of the nanorods and the simplicity of the single-wavelength STED system.

  2. 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...

  3. Observation of high-$p_{T}$ jets in two-photon interactions

    CERN Document Server

    Bartel, Wulfrin; Dittmann, P; Eichler, R; Felst, R; Haidt, Dieter; Krehbiel, H; Meier, K; Naroska, Beate; O'Neill, L H; Steffen, P; Wenninger, Horst; Zhang, Y; Elsen, E E; Helm, M; Petersen, A; Warming, P; Weber, G; Bethke, Siegfried; Drumm, H; Heintze, J; Heinzelmann, G; Hellenbrand, K H; Heuer, R D; Von Krogh, J; Lennert, P; Kawabata, S; Matsumura, H; Nozaki, T; Olsson, J; Rieseberg, H; Wagner, A; Bell, A; Foster, F; Hughes, G; Wriedt, H; Allison, J; Ball, A H; Bamford, G; Barlow, R; Bowdery, C K; Duerdoth, I P; Hassard, J F; King, B T; Loebinger, F K; MacBeth, A A; McCann, H; Mills, H E; Murphy, P G; Stephens, K; Clarke, D; Goddard, M C; Marshall, R; Pearce, G F; Kobayashi, T; Komamiya, S; Koshiba, M; Minowa, M; Nosaki, M; Orito, S; Sato, A; Suda, T; Takeda, H; Totsuka, Y; Watanabe, Y; Yamada, S; Yanagisawa, C

    1981-01-01

    Events with a characteristic two-jet topology have been observed in two-photon interactions. The production cross section is found to be higher than the point-like gamma gamma -qq cross section, which is approached only at transverse momenta larger than 3 GeV/c. (11 refs).

  4. Spectral distribution of the 2S → 1S two-photon transition in atoms ...

    Indian Academy of Sciences (India)

    electron ions. We have measured the spectral ... the photon field operator and ωi is the frequency of transition i. The frequency .... two-photon decay continuum was normalized to the measured spectra at the reduced pho- ton energy of 0.50, i.e., ...

  5. One- and two-photon spectroscopy of a flux qubit coupled to a microscopic defect

    NARCIS (Netherlands)

    Lupa?cu, A.; Bertet, P.; Driessen, E.F.C.; Harmans, C.J.P.M.; Mooij, J.E.

    2009-01-01

    We observed the dynamics of a superconducting flux qubit coupled to an extrinsic quantum system (EQS). The presence of the EQS is revealed by an anticrossing in the spectroscopy of the qubit. The excitation of a two-photon transition to the third excited state of the qubit-EQS system allows us to

  6. Observation of two-photon absorption at UV radiation in ZnS ...

    Indian Academy of Sciences (India)

    2014-02-12

    YAG laser radiation having pulsed duration of 10 ns with the repetition rate of 10 Hz. The measured experimental data have been analysed by using analytical models and two-photon absorption coefficients of the ZnS QDs at ...

  7. Atomic Dipole Squeezing in the Correlated Two-Mode Two-Photon Jaynes-Cummings Model

    Science.gov (United States)

    Dong, Zhengchao; Zhao, Yonglin

    1996-01-01

    In this paper, we study the atomic dipole squeezing in the correlated two-mode two-photon JC model with the field initially in the correlated two-mode SU(1,1) coherent state. The effects of detuning, field intensity and number difference between the two field modes are investigated through numerical calculation.

  8. A study of Two Photon Decays of Charmonium Resonances Formed in Proton Anti-Proton Annihilations

    Energy Technology Data Exchange (ETDEWEB)

    Pedlar, Todd Kristofer [Northwestern Univ., Evanston, IL (United States)

    1999-06-01

    In this dissertation we describe the results of an investigation of the production of charmonium states (ηc, η'c, χ0 and χ2) in Fermilab experiment E835 via antiproton-proton annihilation and their detection via their decay into two photons.

  9. Virtual-pion and two-photon production in pp scattering

    NARCIS (Netherlands)

    Scholten, O; Korchin, AY

    Two-photon production in pp scattering is proposed as a means of studying virtual-pion emission. Such a process is complementary to real-pion emission in pp scattering. The virtual-pion signal is embedded in a background of double-photon bremsstrahlung. We have developed a model to describe this

  10. Search for a Higgs boson decaying into two photons in the CMS ...

    Indian Academy of Sciences (India)

    http://www.ias.ac.in/article/fulltext/pram/079/05/1341-1344. Keywords. CMS; Large Hadron Collider; low-mass Higgs; photons; electromagnetic calorimeter. Abstract. A search for a Higgs boson decaying into two photons in collisions at the LHC at a centre-of-mass energy of 7 TeV is presented. The analysis is performed ...

  11. Cell penetrating silica nanoparticles doped with two-photon absorbing fluorophores

    NARCIS (Netherlands)

    Bertazza, Loris; Celotti, Lucia; Fabbrini, Graziano; Loi, Maria Antonietta; Maggini, Michele; Mancin, Fabrizio; Marcuz, Silvia; Menna, Enzo; Muccini, Michele; Tonellato, Umberto

    2006-01-01

    Organosilica nanoparticles, doped with two-photon absorbing distyrylbenzene derivatives, were prepared and studied as cell staining agents. Two dyes were used, bearing either two peripheral dimethylamino groups or one dimethylamino and one cyano group. Due to the internal charge transfer character

  12. 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.

  13. Development and design of up-to-date laser scanning two-photon microscope using in neuroscience

    Science.gov (United States)

    Doronin, Maxim; Popov, Alexander

    2017-02-01

    Today one of the main areas of application of two-photon microscopy is biology. This is due to the fact that this technique allows to obtain 3D images of tissues due to laser focus change, that is possible due to substantially greater penetration depth on the main wavelength into biological tissues. Self-developed microscopy system provides possibility to service it and modify the structure of microscope depending on highly specialized experimental design and scientific goals. This article may be regarded as a quick reference to laboratory staff who are wishing to develop their own microscopy system for self-service and modernization of the system and in order to save the lab budget.

  14. Two-photon photodetector in a multiquantum well GaAs laser structure at 1.55 microm.

    Science.gov (United States)

    Duchesne, D; Razzari, L; Halloran, L; Morandotti, R; SpringThorpe, A J; Christodoulides, D N; Moss, D J

    2009-03-30

    We report two-photon photocurrent in a GaAs/AlGaAs multiple quantum well laser at 1.55 microm. Using 1ps pulses, a purely quadratic photocurrent is observed. We measure the device efficiency, sensitivity, as well as the two-photon absorption coefficient. The results show that the device has potential for signal processing, autocorrelation and possibly two-photon source applications at sub-Watt power levels.

  15. Metal Complexes for Two-Photon Photodynamic Therapy: A Cyclometallated Iridium Complex Induces Two-Photon Photosensitization of Cancer Cells under Near-IR Light.

    Science.gov (United States)

    McKenzie, Luke K; Sazanovich, Igor V; Baggaley, Elizabeth; Bonneau, Mickaële; Guerchais, Véronique; Williams, J A Gareth; Weinstein, Julia A; Bryant, Helen E

    2017-01-05

    Photodynamic therapy (PDT) uses photosensitizers (PS) which only become cytotoxic upon light-irradiation. Transition-metal complexes are highly promising PS due to long excited-state lifetimes, and high photo-stabilities. However, these complexes usually absorb higher-energy UV/Vis light, whereas the optimal tissue transparency is in the lower-energy NIR region. Two-photon excitation (TPE) can overcome this dichotomy, with simultaneous absorption of two lower-energy NIR-photons populating the same PS-active excited state as one higher-energy photon. We introduce two low-molecular weight, long-lived and photo-stable iridium complexes of the [Ir(N^C)2 (N^N)](+) family with high TP-absorption, which localise to mitochondria and lysosomal structures in live cells. The compounds are efficient PS under 1-photon irradiation (405 nm) resulting in apoptotic cell death in diverse cancer cell lines at low light doses (3.6 J cm(-2) ), low concentrations, and photo-indexes greater than 555. Remarkably 1 also displays high PS activity killing cancer cells under NIR two-photon excitation (760 nm), which along with its photo-stability indicates potential future clinical application. © 2016 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.

  16. Effects of conjugation length and resonance enhancement on two-photon absorption in phenylene–vinylene oligomers

    DEFF Research Database (Denmark)

    Johnsen, Mette; Paterson, M.J.; Arnbjerg, J.

    2008-01-01

    . This phenomenon of the so-called resonance enhancement allows for greater control in obtaining an optimal response when using existing two-photon chromophores, and provides a much-needed guide for the systematic development and efficient use of two-photon singlet oxygen sensitizers....... optical properties in large organic molecules. Specifically, data are provided to indicate that when the frequency of the laser used in the two-photon experiment is nearly-resonant with an allowed one-photon transition, significant increases in the two-photon absorption cross section can be realized...

  17. 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.

  18. Single- and Two-Photon-Induced Processes at the B Factories

    Energy Technology Data Exchange (ETDEWEB)

    Li, Selina Z.; /SLAC

    2012-06-15

    We discuss single- and two-photon-induced processes in e{sup +}e{sup -} annihilations with center-of-mass energy near 10.58 GeV from the BaBar and Belle experiments. In particular, we present experimental results from two-photon physics of {gamma}{gamma} {yields} {pi}{sup 0}{pi}{sup 0} and {gamma}{gamma}* {yields} {pi}{sup 0}. We also review the observation of the Two-Virtual-Photon-Annihilation process (e{sup +}e{sup -} {yields} {rho}{sup 0}{rho}{sup 0} and e{sup +}e{sup -} {yields} {phi}{rho}{sup 0}) and the observation of e{sup +}e{sup -} {yields} {rho}{sup +}{rho}{sup -}, which should be primarily a one virtual photon process, but whose angular distributions may imply potential interference effects.

  19. 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.

  20. Relativistic calculations of the non-resonant two-photon ionization of neutral atoms

    CERN Document Server

    Hofbrucker, Jiri; Fritzsche, Stephan

    2016-01-01

    The non-resonant two-photon one-electron ionization of neutral atoms is studied theoretically in the framework of relativistic second-order perturbation theory and independent particle approximation. In particular, the importance of relativistic and screening effects in the total two-photon ionization cross section is investigated. Detailed computations have been carried out for the K-shell ionization of neutral Ne, Ge, Xe, and U atoms. The relativistic effects significantly decrease the total cross section, for the case of U, for example, they reduce the total cross section by a factor of two. Moreover, we have found that the account for the screening effects of the remaining electrons leads to occurrence of an unexpected minimum in the total cross section at the total photon energies equal to the ionization threshold, for the case of Ne, for example, the cross section drops there by a factor of three.

  1. Holographic multi-focus 3D two-photon polymerization with real-time calculated holograms.

    Science.gov (United States)

    Vizsnyiczai, Gaszton; Kelemen, Lóránd; Ormos, Pál

    2014-10-06

    Two-photon polymerization enables the fabrication of micron sized structures with submicron resolution. Spatial light modulators (SLM) have already been used to create multiple polymerizing foci in the photoresist by holographic beam shaping, thus enabling the parallel fabrication of multiple microstructures. Here we demonstrate the parallel two-photon polymerization of single 3D microstructures by multiple holographically translated foci. Multiple foci were created by phase holograms, which were calculated real-time on an NVIDIA CUDA GPU, and displayed on an electronically addressed SLM. A 3D demonstrational structure was designed that is built up from a nested set of dodecahedron frames of decreasing size. Each individual microstructure was fabricated with the parallel and coordinated motion of 5 holographic foci. The reproducibility and the high uniformity of features of the microstructures were verified by scanning electron microscopy.

  2. Inhibition of two-photon absorption due to dipole-dipole interaction in nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Mahi R. [Department of Physics and Astronomy, University of Western Ontario, London, N6A 3K7 (Canada)], E-mail: msingh@uwo.ca

    2008-07-21

    We have investigated the inhibition of two-photon absorption in photonic crystals doped with an ensemble of four-level nanoparticles. The particles are interacting with one another by the dipole-dipole interaction. Dipoles in nanoparticles are induced by a selected transition. Numerical simulations have been performed for an isotropic photonic crystal. Interesting phenomena have been predicted such as the inhibition of the two-photon absorption due to the dipole-dipole interaction. It has also been found that the inhibition effect can be switched on and off by tuning a decay resonance energy within the energy band of the crystal. A theory of dressed states has been used to explain the results.

  3. Vibrational excitation of hydrogen molecules by two-photon absorption and third-harmonic generation

    Science.gov (United States)

    Miyamoto, Yuki; Hara, Hideaki; Hiraki, Takahiro; Masuda, Takahiko; Sasao, Noboru; Uetake, Satoshi; Yoshimi, Akihiro; Yoshimura, Koji; Yoshimura, Motohiko

    2018-01-01

    We report the coherent excitation of the vibrational state of hydrogen molecules by two-photon absorption and the resultant third-harmonic generation (THG). Parahydrogen molecules cooled by liquid nitrogen are irradiated by mid-infrared nanosecond pulses at 4.8 μm with a nearly Fourier-transform-limited linewidth. The first excited vibrational state of parahydrogen is populated by two-photon absorption of the mid-infrared photons. Because of the narrow linewidth of the mid-infrared pulses, coherence between the ground and excited states is sufficient to induce higher-order processes. Near-infrared photons from the THG are observed at 1.6 μm. The dependence of the intensity of the near-infrared radiation on mid-infrared pulse energy, target pressure, and cell length is determined. We used a simple formula for THG with consideration of realistic experimental conditions to explain the observed results.

  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. Synthesis of Photoresponsive Dual NIR Two-Photon Absorptive [60]Fullerene Triads and Tetrads

    Directory of Open Access Journals (Sweden)

    Long Y. Chiang

    2013-08-01

    Full Text Available Broadband nonlinear optical (NLO organic nanostructures exhibiting both ultrafast photoresponse and a large cross-section of two-photon absorption throughout a wide NIR spectrum may make them suitable for use as nonlinear biophotonic materials. We report here the synthesis and characterization of two C60-(antennax analogous compounds as branched triad C60(>DPAF-C18(>CPAF-C2M and tetrad C60(>DPAF-C18(>CPAF-C2M2 nanostructures. These compounds showed approximately equal extinction coefficients of optical absorption over 400–550 nm that corresponds to near-IR two-photon based excitation wavelengths at 780–1,100 nm. Accordingly, they may be utilized as potential precursor candidates to the active-core structures of photosensitizing nanodrugs for 2γ-PDT in the biological optical window of 800–1,050 nm.

  6. Molecular Tuning of Phenylene-Vinylene Derivatives for Two-Photon Photosensitized Singlet Oxygen Production

    DEFF Research Database (Denmark)

    Nielsen, Christian B.; Arnbjerg, Jacob; Johnsen, Mette

    2009-01-01

    Substituent-dependent features and properties of the sensitizer play an important role in the photosensitized production of singlet oxygen, O2(a1Δg). In this work, we systematically examine the effect of molecular changes in the sensitizer on the efficiency of singlet oxygen production using......, as the sensitizer, oligophenylene-vinylene derivatives designed to optimally absorb light in a nonlinear two-photon process. We demonstrate that one cannot always rely on rule-of-thumb guidelines when attempting to construct efficient two-photon singlet oxygen sensitizers. Rather, as a consequence of behavior...... that can deviate from the norm, a full investigation of the photophysical properties of the system is generally required. For example, it is acknowledged that the introduction of a ketone moiety to the sensitizer chromophore often results in more efficient production of singlet oxygen. However, we show...

  7. Momentum-resolved dynamics of Ar/Cu(1 0 0) interface states probed by time-resolved two-photon photoemission

    Energy Technology Data Exchange (ETDEWEB)

    Rohleder, M; Duncker, K; Berthold, W; Guedde, J; Hoefer, U [Fachbereich Physik und Zentrum fuer Materialwissenschaften, Philipps-Universitaet, D-35032 Marburg (Germany)

    2005-04-01

    The electron dynamics of buried Ar/Cu(1 0 0) image-potential states was investigated by time-resolved two-photon photoemission (2PPE) as a function of parallel momentum. The first interface state shows a parabolic dispersion with an effective mass of 0.6. Its lifetime of 110 fs at the {gamma}-bar-point decreases with increasing parallel momentum. The momentum dependence of the decay can be understood by intra- and inter-band decay processes mediated by Cu electrons, just as the decay of image-potential states on the clean Cu(1 0 0) surface.

  8. Aggregation induced enhanced emission of conjugated dendrimers with a large intrinsic two-photon absorption cross-section

    NARCIS (Netherlands)

    Xu, Bin; Zhang, Jibo; Fang, Honghua; Ma, Suqian; Chen, Qidai; Sun, Hongbo; Im, Chan; Tian, Wenjing

    2014-01-01

    Organic nonlinear optical materials combining high luminescence quantum yields and large two-photon absorption cross-sections are attractive for both fundamental research and practical applications, such as up-converted lasers and two-photon fluorescence microscopy. Herein, we reported a series of

  9. 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).

  10. Resummation of target mass corrections in two-photon processes: twist-two sector

    Science.gov (United States)

    Belitsky, A. V.; Müller, D.

    2001-05-01

    We develop a formalism for the resummation of target mass corrections in off-forward two-photon amplitudes given by a chronological product of electromagnetic currents, arising in, e.g., deeply virtual Compton scattering. The method is based on a relation of composite operators with a definite twist to harmonic tensors, which form an irreducible representation of the Lorentz group. We give an application of the framework for the matrix elements of twist-two operators.

  11. Development and design of advanced two-photon microscope used in neuroscience

    Science.gov (United States)

    Doronin, M. S.; Popov, A. V.

    2016-08-01

    This work represents the real steps to development and design advanced two-photon microscope by efforts of laboratory staff. Self-developed microscopy system provides possibility to service it and modify the structure of microscope depending on highly specialized experimental design and scientific goals. We are presenting here module-based microscopy system which provides an opportunity to looking for new applications of this setup depending on laboratories needs using with galvo and resonant scanners.

  12. Recent Results on Two-Photon Physics at BaBar

    Energy Technology Data Exchange (ETDEWEB)

    Druzhinin, Vladimir P.; /Novosibirsk, IYF

    2011-12-02

    The authors present measurements of the {gamma}{gamma}* {yields} {pi}{sup 0} transition form factor for the momentum transfer range Q{sup 2} = 4-40 GeV{sup 2} and the {gamma}{gamma}* {yields} {eta}{sub c} transition form factor for the range Q{sup 2} = 2-50 GeV{sup 2}. The results of the measurement of the {eta}{sub c} mass, total and two-photon widths are also presented.

  13. Polarization-resolved two-photon luminescence microscopy of V-groove arrays

    DEFF Research Database (Denmark)

    Beermann, J.; Novikov, S. M.; Holmgaard, T.

    2012-01-01

    Using two-photon luminescence (TPL) microscopy and local reflection spectroscopy we investigate electromagnetic field enhancement effects from a mu m-sized composition of 450-nm-deep V-grooves milled by focused ion beam in a thick gold film and assembled to feature, within the same structure...... obtained to evaluation of local field enhancements using TPL microscopy, especially when investigating extended structures exhibiting different radiation channels, are discussed. (C)2011 Optical Society of America...

  14. Bandgap Engineering of Double Perovskites for One- and Two-photon Water Splitting

    DEFF Research Database (Denmark)

    Castelli, Ivano Eligio; Thygesen, Kristian Sommer; Jacobsen, Karsten Wedel

    2013-01-01

    Computational screening is becoming increasingly useful in the search for new materials. We are interested in the design of new semiconductors to be used for light harvesting in a photoelectrochemical cell. In the present paper, we study the double perovskite structures obtained by combining 46 s...... in the volume has an effect on the size of the bandgap. In addition, we suggest some new candidate materials that can be used as photocatalysts in one- and two-photon water splitting devices....

  15. Higgs boson decay into two photons in an electromagnetic background field

    DEFF Research Database (Denmark)

    Nielsen, N. K.

    2014-01-01

    The amplitude for Higgs boson decay into two photons in a homogeneous and time-independent magnetic field is investigated by proper-time regularization in a gauge-invariant manner and is found to be singular at large field values. The singularity is caused by the component of the charged vector...... boson field that is tachyonic in a strong magnetic field. Also, tools for the computation of the amplitude in a more general electromagnetic background are developed....

  16. Search for a Higgs boson decaying into two photons in the CMS ...

    Indian Academy of Sciences (India)

    2012-11-15

    Nov 15, 2012 ... H → γγ is one of the most promising channels for Higgs discovery in the very low mass region. Despite its small branching ratio of about 0.2% in this region of interest, the. H → γγ decay channel provides a clean final-state topology, since the invariant mass of the two photons can be reconstructed with great ...

  17. New cubic perovskites for one- and two-photon water splitting using the computational materials repository

    DEFF Research Database (Denmark)

    Castelli, Ivano Eligio; Landis, David; Thygesen, Kristian Sommer

    2012-01-01

    screening of around 19 000 oxides, oxynitrides, oxysulfides, oxyfluorides, and oxyfluoronitrides in the cubic perovskite structure with PEC applications in mind. We address three main applications: light absorbers for one- and two-photon water splitting and high-stability transparent shields to protect...... against corrosion. We end up with 20, 12, and 15 different combinations of oxides, oxynitrides and oxyfluorides, respectively, inviting further experimental investigation....

  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. Rapid Prototyping of Chemical Microsensors Based on Molecularly Imprinted Polymers Synthesized by Two-Photon Stereolithography.

    Science.gov (United States)

    Gomez, Laura Piedad Chia; Spangenberg, Arnaud; Ton, Xuan-Anh; Fuchs, Yannick; Bokeloh, Frank; Malval, Jean-Pierre; Tse Sum Bui, Bernadette; Thuau, Damien; Ayela, Cédric; Haupt, Karsten; Soppera, Olivier

    2016-07-01

    Two-photon stereolithography is used for rapid prototyping of submicrometre molecularly imprinted polymer-based 3D structures. The structures are evaluated as chemical sensing elements and their specific recognition properties for target molecules are confirmed. The 3D design capability is exploited and highlighted through the fabrication of an all-organic molecularly imprinted polymeric microelectromechanical sensor. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  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....

  1. Ultrashort coherence times in partially polarized stationary optical beams measured by two-photon absorption.

    Science.gov (United States)

    Shevchenko, Andriy; Roussey, Matthieu; Friberg, Ari T; Setälä, Tero

    2015-11-30

    We measure the recently introduced electromagnetic temporal degree of coherence of a stationary, partially polarized, classical optical beam. Instead of recording the visibility of intensity fringes, the spectrum, or the polarization characteristics, we introduce a novel technique based on two-photon absorption. Using a Michelson interferometer equipped with polarizers and a specific GaAs photocount tube, we obtain the two fundamental quantities pertaining to the fluctuations of light: the degree of coherence and the degree of polarization. We also show that the electromagnetic intensity-correlation measurements with two-photon absorption require that the polarization dynamics, i.e., the time evolution of the instantaneous polarization state, is properly taken into account. We apply the technique to unpolarized and polarized sources of amplified spontaneous emission (Gaussian statistics) and to a superposition of two independent, narrow-band laser beams of different mid frequencies (non-Gaussian statistics). For these two sources femtosecond-range coherence times are found that are in good agreement with the traditional spectral measurements. Although previously employed for laser pulses, two-photon absorption provides a new physical principle to study electromagnetic coherence phenomena in classical and quantum continuous-wave light at extremely short time scales.

  2. 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.

  3. Two-Photon Excitation of Conjugated Molecules in Solution: Spectroscopy and Excited-State Dynamics

    Science.gov (United States)

    Elles, Christopher G.; Houk, Amanda L.; de Wergifosse, Marc; Krylov, Anna

    2017-06-01

    We examine the two-photon absorption (2PA) spectroscopy and ultrafast excited-state dynamics of several conjugated molecules in solution. By controlling the relative wavelength and polarization of the two photons, the 2PA measurements provide a more sensitive means of probing the electronic structure of a molecule compared with traditional linear absorption spectra. We compare experimental spectra of trans-stilbene, cis-stilbene, and phenanthrene in solution with the calculated spectra of the isolated molecules using EOM-EE-CCSD. The calculated spectra show good agreement with the low-energy region of the experimental spectra (below 6 eV) after suppressing transitions with strong Rydberg character and accounting for solvent and method-dependent shifts of the valence transitions. We also monitor the excited state dynamics following two-photon excitation to high-lying valence states of trans-stilbene up to 6.5 eV. The initially excited states rapidly relax to the lowest singlet excited state and then follow the same reaction path as observed following direct one-photon excitation to the lowest absorption band at 4.0 eV.

  4. 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.

  5. Design and Analysis of XOR Logic Gate Based on Two-Photon Absorption

    Directory of Open Access Journals (Sweden)

    Nazir N.S.

    2016-01-01

    Full Text Available Two-photon absorption with high intensity pump beam occurs in a SOA depends on fast phase change of a weak probe signal. This work analysed optical XOR logic function using two-photon absorption induced fast phase change. A rate equation for SOA and both input data signals A and B with high intensity, configured in the form of MZI has also been proved. The model shows that XOR operation at 10 Gb/s with good signal to noise ratio is obtained with high input intensities. The result on the generation of XOR indicates that operations on 10 Gb/s with a high signal to noise ratio can easily be implemented. The average input power into the SOA is 20 dbm corresponding to the peak power of 5.5 dBm at 10 Gb/s when the width of the input pulse 3.6 ps. The short narrow pulse width is utilised in the study for stronger effect of two-photon absorption.

  6. A Study of Two-Prong Two-Photon Events at the Z0 Resonance

    Energy Technology Data Exchange (ETDEWEB)

    Kowalski, L.

    2004-08-09

    A study of electron-positron scattering leading to two electron-positron pairs via a two-photon interaction has been carried out at the Stanford Linear Accelerator Center (SLAC). The case where one pair is observed in the detector was investigated. These events were produced at the SLAC Linear Collider (SLC) operating in the center-of-mass energy range from 89.2 to 9.30 GeV. The data was collected using the Mark II detector. Two-photon interactions are described by the theory of quantum electrodynamics (QED). Such processes can be a significant background to new particle searches; consequently, and understanding of their production is imperative. A deviation from the event rate predicted by QED might indicate the existence of new particles. The event rate may also be useful as a luminosity monitor during data collection. The data sample from the Mark II is searched for events which have features indicative of two-photon events. For comparison with theory, the Berends, Daverveldt, and Keiss Monte Carlo event generator is used to simulate events according to QED theory. The data is compared to the theoretical predictions. Given the low event statistics from the SLC data run, the results are consistent with the QED theoretical prediction. However, due to the low statistics, this measurement cannot be used to indicate non-deviation from QED predictions.

  7. Two-photon production of neutrinos as a factor in supernova formation

    CERN Document Server

    Campbell, J A

    1973-01-01

    The reaction in which a neutrino-antineutrino pair is produced by the annihilation of two photons is suggested as having a strong influence on the formation of supernovae. Results of calculations in two different non-local theories of the weak interaction are given, which become identical if the different heavy particles in the two theories have a common mass just below 20 GeV. Use of the consequent single expression for neutrino luminosity in astrophysical models is recommended because of its unique ability to distinguish between local and non-local theories in particle physics, and to indicate whether there is a chance of removing divergences from non-local theories. Specific effects of the two-photon reaction on general models of the formation of supernovae are pointed out. In particular, the maximum mass of a star which can turn into a supernova may be quite low, and the fraction of mass ejected as cosmic rays may be increased significantly over the fraction in models without two-photon annihilation.

  8. Two-Photon-Excited Silica and Organosilica Nanoparticles for Spatiotemporal Cancer Treatment.

    Science.gov (United States)

    Croissant, Jonas G; Zink, Jeffrey I; Raehm, Laurence; Durand, Jean-Olivier

    2018-01-18

    Coherent two-photon-excited (TPE) therapy in the near-infrared (NIR) provides safer cancer treatments than current therapies lacking spatial and temporal selectivities because it is characterized by a 3D spatial resolution of 1 µm 3 and very low scattering. In this review, the principle of TPE and its significance in combination with organosilica nanoparticles (NPs) are introduced and then studies involving the design of pioneering TPE-NIR organosilica nanomaterials are discussed for bioimaging, drug delivery, and photodynamic therapy. Organosilica nanoparticles and their rich and well-established chemistry, tunable composition, porosity, size, and morphology provide ideal platforms for minimal side-effect therapies via TPE-NIR. Mesoporous silica and organosilica nanoparticles endowed with high surface areas can be functionalized to carry hydrophobic and biologically unstable two-photon absorbers for drug delivery and diagnosis. Currently, most light-actuated clinical therapeutic applications with NPs involve photodynamic therapy by singlet oxygen generation, but low photosensitizing efficiencies, tumor resistance, and lack of spatial resolution limit their applicability. On the contrary, higher photosensitizing yields, versatile therapies, and a unique spatial resolution are available with engineered two-photon-sensitive organosilica particles that selectively impact tumors while healthy tissues remain untouched. Patients suffering pathologies such as retinoblastoma, breast, and skin cancers will greatly benefit from TPE-NIR ultrasensitive diagnosis and therapy. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Molecular tuning of phenylene-vinylene derivatives for two-photon photosensitized singlet oxygen production.

    Science.gov (United States)

    Nielsen, Christian B; Arnbjerg, Jacob; Johnsen, Mette; Jorgensen, Mikkel; Ogilby, Peter R

    2009-12-04

    Substituent-dependent features and properties of the sensitizer play an important role in the photosensitized production of singlet oxygen, O(2)(a(1)Delta(g)). In this work, we systematically examine the effect of molecular changes in the sensitizer on the efficiency of singlet oxygen production using, as the sensitizer, oligophenylene-vinylene derivatives designed to optimally absorb light in a nonlinear two-photon process. We demonstrate that one cannot always rely on rule-of-thumb guidelines when attempting to construct efficient two-photon singlet oxygen sensitizers. Rather, as a consequence of behavior that can deviate from the norm, a full investigation of the photophysical properties of the system is generally required. For example, it is acknowledged that the introduction of a ketone moiety to the sensitizer chromophore often results in more efficient production of singlet oxygen. However, we show here that the introduction of a carbonyl into a given phenylene-vinylene can, rather, have adverse effects on the yield of singlet oxygen produced. Using these molecules, we show that care must also be exercised when using qualitative symmetry-derived arguments to predict the relationship between one-and two-photon absorption spectra.

  10. 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.

  11. Effects of cholesterol on plasma membrane lipid order in MCF-7 cells by two-photon microscopy

    Science.gov (United States)

    Zeng, Yixiu; Chen, Jianling; Yang, Hongqin; Wang, Yuhua; Li, Hui; Xie, Shusen

    2014-09-01

    Lipid rafts are cholesterol- and glycosphingolipids- enriched microdomains on plasma membrane surface of mammal cells, involved in a variety of cellular processes. Depleting cholesterol from the plasma membrane by drugs influences the trafficking of lipid raft markers. Optical imaging techniques are powerful tools to study lipid rafts in live cells due to its noninvasive feature. In this study, breast cancer cells MCF-7 were treated with different concentrations of MβCD to deplete cholesterol and an environmentally sensitive fluorescence probe, Laurdan was loaded to image lipid order by two-photon microscopy. The generalized polarization (GP) values were calculated to distinguish the lipid order and disorder phase. GP images and GP distributions of native and cholesterol-depleted MCF-7 cells were obtained. Our results suggest that even at low concentration (0.5 mM) of MβCD, the morphology of the MCF-7 cells changes. Small high GP areas (lipid order phase) decrease more rapidly than low GP areas (lipid disorder phase), indicating that lipid raft structure was altered more severely than nonraft domains. The data demonstrates that cholesterol dramatically affect raft coverage and plasma membrane fluidity in living cells.

  12. An organic dye with very large Stokes-shift and broad tunability of fluorescence: Potential two-photon probe for bioimaging and ultra-sensitive solid-state gas sensor

    Energy Technology Data Exchange (ETDEWEB)

    He, Tingchao; Tian, Xiaoqing; Lin, Xiaodong, E-mail: linxd@szu.edu.cn, E-mail: hdsun@ntu.edu.sg [College of Physics Science and Technology, Shenzhen University, Shenzhen 518060 (China); Wang, Yue; Zhao, Xin; Sun, Handong, E-mail: linxd@szu.edu.cn, E-mail: hdsun@ntu.edu.sg [Division of Physics and Applied Physics, and Centre for Disruptive Photonic Technologies (CDPT), School of Physical and Mathematical Sciences, Nanyang Technological University, 21 Nanyang Link, Singapore 637371 (Singapore); Gao, Yang; Grimsdale, Andrew C. [School of Materials Science and Engineering, Nanyang Technological University, Singapore 639798 (Singapore)

    2016-01-04

    Light-emitting nonlinear optical molecules, especially those with large Stokes shifts and broad tunability of their emission wavelength, have attracted considerable attention for various applications including biomedical imaging and fluorescent sensors. However, most fluorescent chromophores have only limited potential for such applications due to small Stokes shifts, narrow tunability of fluorescence emissions, and small optical nonlinearity in highly polar solvents. In this work, we demonstrate that a two-photon absorbing stilbene chromophore exhibits a large two-photon absorption action cross-section (ηδ = 320 GM) in dimethylsulfoxide (DMSO) and shows broad fluorescence tunability (125 nm) by manipulating the polarity of the surrounding medium. Importantly, a very large Stokes shift of up to 227 nm is achieved in DMSO. Thanks to these features, this chromophore can be utilized as a two-photon probe for bioimaging applications and in an ultrasensitive solid-state gas detector.

  13. Three-Dimensional Segmentation and Quantitative Measurement of the Aqueous Outflow System of Intact Mouse Eyes Based on Spectral Two-Photon Microscopy Techniques.

    Science.gov (United States)

    Zhang, Xianzeng; Liu, Nenrong; Mak, Peng Un; Pun, Sio Hang; Vai, Mang I; Masihzadeh, Omid; Kahook, Malik Y; Lei, Tim C; Ammar, David A

    2016-06-01

    To visualize and quantify the three-dimensional (3D) spatial relationships of the structures of the aqueous outflow system (AOS) within intact enucleated mouse eyes using spectral two-photon microscopy (TPM) techniques. Spectral TPM, including two-photon autofluorescence (TPAF) and second-harmonic generation (SHG), were used to image the small structures of the AOS within the limbal region of freshly enucleated mouse eyes. Long infrared excitation wavelengths (930 nm) were used to reduce optical scattering and autofluorescent background. Image stacks were collected for 3D image rendering and structural segmentation. For anatomical reference, vascular perfusion with fluorescent-conjugated dextran (150 KDa) and trans-corneal perfusion with 0.1 μm fluorescent polystyrene beads were separately performed to identify the episcleral veins (EV) and the trabecular meshwork (TM) and Schlemm's canal (SC), respectively. Three-dimensional rendering and segmentation of spectral two-photon images revealed detailed structures of the AOS, including SC, collector channels (CC), and aqueous veins (AV). The collagen of the TM was detected proximal to SC. The long and short axes of the SC were 82.2 ± 22.2 μm and 6.7 ± 1.4 μm. The diameters of the CC averaged 25.6 ± 7.9 μm where they originated from the SC (ostia), enlarged to 34.1 ± 13.1 μm at the midpoint, and narrowed to 18.3 ± 4.8 μm at the junction of the AV. The diameter of the AV averaged 12.5 ± 3.4 μm. Spectral TPM can be used to reconstruct and measure the spatial relationships of both large and small AOS structures, which will allow for better understanding of distal aqueous outflow dynamics.

  14. Fabrication of microscale medical devices by two-photon polymerization with multiple foci via a spatial light modulator.

    Science.gov (United States)

    Gittard, Shaun D; Nguyen, Alexander; Obata, Kotaro; Koroleva, Anastasia; Narayan, Roger J; Chichkov, Boris N

    2011-11-01

    Two-photon polymerization is an appealing technique for producing microscale devices due to its flexibility in producing structures with a wide range of geometries as well as its compatibility with materials suitable for biomedical applications. The greatest limiting factor in widespread use of two-photon polymerization is the slow fabrication times associated with line-by-line, high-resolution structuring. In this study, a recently developed technology was used to produce microstructures by two-photon polymerization with multiple foci, which significantly reduces the production time. Computer generated hologram pattern technology was used to generate multiple laser beams in controlled positions from a single laser. These multiple beams were then used to simultaneously produce multiple microstructures by two-photon polymerization. Arrays of micro-Venus structures, tissue engineering scaffolds, and microneedle arrays were produced by multifocus two-photon polymerization. To our knowledge, this work is the first demonstration of multifocus two-photon polymerization technology for production of a functional medical device. Multibeam fabrication has the potential to greatly improve the efficiency of two-photon polymerization production of microscale devices such as tissue engineering scaffolds and microneedle arrays.

  15. Two-photon absorption induced by electric field gradient of optical near-field and its application to photolithography

    Science.gov (United States)

    Yamaguchi, Maiku; Nobusada, Katsuyuki; Kawazoe, Tadashi; Yatsui, Takashi

    2015-05-01

    An electric field gradient is an inherent property of the optical near-field (ONF). We investigated its effect on electron excitation in a quantum dot via model calculations combining a density matrix formalism and a classical Lorentz model. The electric field gradient of the ONF was found to cause two-photon absorption by an unusual mechanism. Furthermore, the absorption exhibits a nonmonotonic dependence on the spatial arrangement of the nanosystem, completely different from that of conventional two-photon absorption induced by an intense electric field. The present two-photon absorption process was verified in a previous experimental observation by reinterpreting the results of ONF photolithography.

  16. Two-photon absorption induced by electric field gradient of optical near-field and its application to photolithography

    Energy Technology Data Exchange (ETDEWEB)

    Yamaguchi, Maiku; Kawazoe, Tadashi; Yatsui, Takashi [Department of Electrical Engineering and Information Systems, Graduate School of Engineering, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-8656 (Japan); Nobusada, Katsuyuki, E-mail: nobusada@ims.ac.jp [Department of Theoretical and Computational Molecular Science, Institute for Molecular Science, Myodaiji, Okazaki 444-8585 (Japan)

    2015-05-11

    An electric field gradient is an inherent property of the optical near-field (ONF). We investigated its effect on electron excitation in a quantum dot via model calculations combining a density matrix formalism and a classical Lorentz model. The electric field gradient of the ONF was found to cause two-photon absorption by an unusual mechanism. Furthermore, the absorption exhibits a nonmonotonic dependence on the spatial arrangement of the nanosystem, completely different from that of conventional two-photon absorption induced by an intense electric field. The present two-photon absorption process was verified in a previous experimental observation by reinterpreting the results of ONF photolithography.

  17. Watching Electrons Transfer from Metals to Insulators using Two Photon Photoemission

    Energy Technology Data Exchange (ETDEWEB)

    Johns, James E. [Univ. of California, Berkeley, CA (United States)

    2010-05-01

    Ultrafast angle-resolved two photon photoemission was used to study the dynamics and interfacial band structure of ultrathin films adsorbed onto Ag(111). Studies focused on the image potential state (IPS) in each system as a probe for measuring changes in electronic behavior in differing environments. The energetics and dynamics of the IPS at the toluene/Ag(111) interface are strongly dependent upon coverage. For a single monolayer, the first IPS is bound by 0.81 eV below the vacuum level and has a lifetime of 50 femtoseconds (fs). Further adsorption of toluene creates islands of toluene with an exposed wetting layer underneath. The IPS is then split into two peaks, one corresponding to the islands and one corresponding to the monolayer. The wetting layer IPS shows the same dynamics as the monolayer, while the lifetime of the islands increases exponentially with increasing thickness. Furthermore, the island IPS transitions from delocalized to localized within 500 fs, and electrons with larger parallel momenta decay much faster. Attempts were made using a stochastic model to extract the rates of localization and intraband cooling at differing momenta. In sexithiophene (6T) and dihexyl-sexithiophene (DH6T), the IPS was used as a probe to see if the nuclear motion of spectating side chains can interfere with molecular conduction. The energy and band mass of the IPS was measured for 6T and two geometries of DH6T on Ag(111). Electrons injected into the thicker coverages of DH6T grew exponentially heavier until they were completely localized by 230 fs, while those injected into 6T remained nearly free electron like. Based off of lifetime arguments and the density of defects, the most likely cause for the mass enhancement of the IPS in this system is small polaron formation caused by coupling of the electron to vibrations of the alkyl substituents. The energetic relaxation of the molecular adsorbate was also measured to be 20 meV/100 fs for the DH6T, and 0 meV/100 fs for

  18. Polarization dependence of two-photon transition intensities in rare-earth doped crystals

    Energy Technology Data Exchange (ETDEWEB)

    Le Nguyen, An-Dien [Univ. of California, Berkeley, CA (United States)

    1996-05-01

    A polarization dependence technique has been developed as a tool to investigate phonon scattering (PS), electronic Raman scattering (ERS), and two-photon absorption (TPA) transition intensities in vanadate and phosphate crystals. A general theory for the polarization dependence (PD) of two-photon transition intensities has been given. Expressions for the polarization dependent behavior of two-photon transition intensities have been tabulated for the 32 crystallographic point groups. When the wavefunctions for the initial and final states of a rare-earth doped in crystals are known, explicit PD expressions with no unknown parameters can be obtained. A spectroscopic method for measuring and interpreting phonon and ERS intensities has been developed to study PrVO4, NdVO4, ErVO4, and TmVO4 crystals. Relative phonon intensities with the polarization of the incident and scattered light arbitrarily varied were accurately predicted and subsequently used for alignment and calibration in ERS measurements in these systems for the first time. Since ERS and PS intensities generally follow different polarization curves as a function of polar angles, the two can be uniquely identified by comparing their respective polarization behavior. The most crucial application of the technique in ERS spectroscopy is the establishment of a stringent test for the Axe theory. For the first time, the F1/F2 ratio extracted from the experimental fits of the ERS intensities were compared with those predicted by theories which include both the second- and third-order contributions. Relatively good agreement between the fitted values of F1/F2 and the predicted values using the second-order theory has been found.

  19. Sub-millisecond optogenetic control of neuronal firing with two-photon holographic photoactivation of Chronos.

    Science.gov (United States)

    Ronzitti, E; Conti, R; Zampini, V; Tanese, D; Foust, A J; Klapoetke, N; Boyden, E S; Papagiakoumou, E; Emiliani, V

    2017-10-02

    Optogenetic neuronal network manipulation promises to unravel a long-standing mystery in neuroscience: how does microcircuit activity causally relate to behavioral and pathological states? The challenge to evoke spikes with high spatial and temporal complexity necessitates further joint development of light-delivery approaches and custom opsins. Two-photon light-targeting strategies demonstrated, in-depth generation of action potentials in photosensitive neurons both in-vitro and in-vivo, but thus far lack the temporal precision necessary to induce precisely timed spiking events. Here, we show that efficient current integration enabled by two-photon holographic amplified laser illumination of Chronos, a highly light-sensitive and fast opsin, can evoke spikes with submillisecond precision and repeated firing up to 100 Hz in brain slices from Swiss male mice. These results pave the way for optogenetic manipulation with the spatial and temporal sophistication necessary to mimic natural microcircuit activity.SIGNIFICANT STATEMENTTo reveal causal links between neuronal activity and behavior, it is necessary to develop experimental strategies to induce spatially and temporally sophisticated perturbation of network microcircuits. Two-photon computer generated holography (2P-CGH) recently demonstrated three-dimensional optogenetic control of selected pools of neurons with single-cell accuracy in-depth in the brain. Here we show that exciting the fast opsin Chronos with amplified laser 2P-CGH enables cellular-resolution targeting with unprecedented temporal control, driving spiking up to 100 Hz with submillisecond onset precision, using low laser power densities. This system achieves a unique combination of spatial flexibility and temporal precision needed to optogenetically pattern inputs that mimic natural neuronal network activity patterns. Copyright © 2017 the authors.

  20. Muon-Pair and Tau-Pair Production in Two-Photon Collisions at LEP

    CERN Document Server

    Achard, P.; Aguilar-Benitez, M.; Alcaraz, J.; Alemanni, G.; Allaby, J.; Aloisio, A.; Alviggi, M.G.; Anderhub, H.; Andreev, Valery 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.; Barillere, R.; Bartalini, P.; Basile, M.; Batalova, N.; Battiston, R.; Bay, A.; Becattini, F.; Becker, U.; Behner, F.; Bellucci, L.; Berbeco, R.; Berdugo, J.; Berges, P.; Bertucci, B.; Betev, B.L.; Biasini, M.; Biglietti, M.; Biland, A.; Blaising, J.J.; Blyth, S.C.; Bobbink, G.J.; Bohm, A.; Boldizsar, L.; Borgia, B.; Bottai, S.; Bourilkov, D.; Bourquin, M.; Braccini, S.; Branson, J.G.; Brochu, F.; Burger, J.D.; Burger, W.J.; Cai, X.D.; Capell, M.; Cara Romeo, G.; Carlino, G.; Cartacci, A.; Casaus, J.; Cavallari, F.; Cavallo, N.; Cecchi, C.; Cerrada, M.; Chamizo, M.; Chang, Y.H.; Chemarin, M.; Chen, A.; Chen, G.; Chen, G.M.; Chen, H.F.; Chen, H.S.; Chiefari, G.; Cifarelli, L.; Cindolo, F.; Clare, I.; Clare, R.; Coignet, G.; Colino, N.; Costantini, S.; de la Cruz, B.; Cucciarelli, S.; van Dalen, J.A.; de Asmundis, R.; Deglon, P.; Debreczeni, J.; Degre, A.; Dehmelt, K.; Deiters, K.; della Volpe, D.; Delmeire, E.; Denes, P.; DeNotaristefani, F.; De Salvo, A.; Diemoz, M.; Dierckxsens, M.; Dionisi, C.; Dittmar, M.; Doria, A.; Dova, M.T.; Duchesneau, D.; Duda, M.; Echenard, B.; Eline, A.; El Hage, A.; El Mamouni, H.; Engler, A.; Eppling, F.J.; Extermann, P.; Falagan, M.A.; Falciano, S.; Favara, A.; Fay, J.; Fedin, O.; Felcini, M.; Ferguson, T.; Fesefeldt, H.; Fiandrini, E.; Field, J.H.; Filthaut, F.; Fisher, P.H.; Fisher, W.; Fisk, I.; Forconi, G.; Freudenreich, K.; Furetta, C.; Galaktionov, Iouri; Ganguli, S.N.; Garcia-Abia, Pablo; Gataullin, M.; Gentile, S.; Giagu, S.; Gong, Z.F.; Grenier, Gerald Jean; Grimm, O.; Gruenewald, M.W.; Guida, M.; van Gulik, R.; Gupta, V.K.; Gurtu, A.; Gutay, L.J.; Haas, D.; Hatzifotiadou, D.; Hebbeker, T.; Herve, Alain; Hirschfelder, J.; Hofer, H.; Hohlmann, M.; Holzner, G.; Hou, S.R.; Hu, Y.; Jin, B.N.; Jones, Lawrence W.; de Jong, P.; Josa-Mutuberria, I.; Kaur, M.; Kienzle-Focacci, M.N.; Kim, J.K.; Kirkby, Jasper; Kittel, W.; Klimentov, A.; Konig, A.C.; Kopal, M.; Koutsenko, V.; Kraber, M.; Kraemer, R.W.; Kruger, A.; Kunin, A.; Ladron 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.; Levtchenko, P.; Li, C.; Likhoded, S.; Lin, C.H.; Lin, W.T.; Linde, F.L.; Lista, L.; Liu, Z.A.; Lohmann, W.; Longo, E.; Lu, Y.S.; Luci, C.; Luminari, L.; Lustermann, W.; Ma, W.G.; Malgeri, L.; Malinin, A.; Mana, C.; Mans, J.; Martin, J.P.; Marzano, F.; Mazumdar, K.; McNeil, R.R.; Mele, S.; Merola, L.; Meschini, M.; Metzger, W.J.; Mihul, A.; Milcent, H.; Mirabelli, G.; Mnich, J.; Mohanty, G.B.; Muanza, G.S.; Muijs, A.J.M.; Musicar, B.; Musy, M.; Nagy, S.; Natale, S.; Napolitano, M.; Nessi-Tedaldi, F.; Newman, H.; Nisati, A.; Novak, T.; Kluge, Hannelies; Ofierzynski, R.; Organtini, G.; Pal, I.; Palomares, C.; Paolucci, P.; Paramatti, R.; Passaleva, G.; Patricelli, S.; Paul, Thomas Cantzon; Pauluzzi, M.; Paus, C.; Pauss, F.; Pedace, M.; Pensotti, S.; Perret-Gallix, D.; Petersen, B.; Piccolo, D.; Pierella, F.; Pioppi, M.; Piroue, P.A.; Pistolesi, E.; Plyaskin, V.; Pohl, M.; Pojidaev, V.; Pothier, J.; Prokofev, D.; Quartieri, J.; Rahal-Callot, G.; Rahaman, Mohammad Azizur; Raics, P.; Raja, N.; Ramelli, R.; Rancoita, P.G.; Ranieri, R.; Raspereza, A.; Razis, P.; Ren, D.; Rescigno, M.; Reucroft, S.; Riemann, S.; Riles, Keith; Roe, B.P.; Romero, L.; Rosca, A.; Rosemann, C.; Rosenbleck, C.; Rosier-Lees, S.; Roth, Stefan; Rubio, J.A.; Ruggiero, G.; Rykaczewski, H.; Sakharov, A.; Saremi, S.; Sarkar, S.; Salicio, J.; Sanchez, E.; Schafer, C.; Schegelsky, V.; Schopper, H.; Schotanus, D.J.; Sciacca, C.; Servoli, L.; Shevchenko, S.; Shivarov, N.; Shoutko, V.; Shumilov, E.; Shvorob, A.; Son, D.; Souga, C.; Spillantini, P.; Steuer, M.; Stickland, D.P.; Stoyanov, B.; Straessner, A.; Sudhakar, K.; Sultanov, G.; Sun, L.Z.; Sushkov, S.; Suter, H.; Swain, J.D.; Szillasi, Z.; Tang, X.W.; Tarjan, P.; Tauscher, L.; Taylor, L.; Tellili, B.; Teyssier, D.; Timmermans, Charles; Ting, Samuel C.C.; Ting, S.M.; Tonwar, S.C.; Toth, J.; Tully, C.; Tung, K.L.; Ulbricht, J.; Valente, E.; Van de Walle, R.T.; Vasquez, R.; Veszpremi, V.; Vesztergombi, G.; Vetlitsky, I.; Vicinanza, D.; Viertel, G.; Villa, S.; Vivargent, M.; Vlachos, S.; Vodopianov, I.; Vogel, H.; Vogt, H.; Vorobev, I.; Vorobyov, A.A.; Wadhwa, M.; Wang, Q.Wang X.L.; Wang, Z.M.; Weber, M.; 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, An.; Zalite, Yu.; Zhang, Z.P.; Zhao, J.; Zhu, G.Y.; Zhu, R.Y.; Zhuang, H.L.; Zichichi, A.; Zimmermann, B.; Zoller, M.

    2004-01-01

    The QED processes e^+ e^- -> e^+ e^- \\mu^+ \\mu^- and e^+ e^- -> e^+ e^- \\tau^+ \\tau^- are studied with the L3 detector at LEP using an untagged data sample collected at centre-of-mass energies 161 GeV \\mu^+\\mu^- process is also measured as a function of the two-photon centre-of-mass energy for 3 GeV < W_{\\gamma\\gamma} < 40 GeV. Good agreement is found between these measurements and the O(\\alpha^4) QED expectations. In addition, limits on the anomalous magnetic and electric dipole moments of the tau lepton are extracted.

  1. $\\Lambda$ and $\\Sigma^{0}$ Pair Production in Two-Photon Collisions at LEP

    CERN Document Server

    Achard, P.; Aguilar-Benitez, M.; Alcaraz, J.; Alemanni, G.; Allaby, J.; Aloisio, A.; Alviggi, M.G.; Anderhub, H.; Andreev, Valery 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.; Barillere, R.; Bartalini, P.; Basile, M.; Batalova, N.; Battiston, R.; Bay, A.; Becattini, F.; Becker, U.; Behner, F.; Bellucci, L.; Berbeco, R.; Berdugo, J.; Berges, P.; Bertucci, B.; Betev, B.L.; Biasini, M.; Biglietti, M.; Biland, A.; Blaising, J.J.; Blyth, S.C.; Bobbink, G.J.; Bohm, A.; Boldizsar, L.; Borgia, B.; Bottai, S.; Bourilkov, D.; Bourquin, M.; Braccini, S.; Branson, J.G.; Brochu, F.; Burger, J.D.; Burger, W.J.; Cai, X.D.; Capell, M.; Cara Romeo, G.; Carlino, G.; Cartacci, A.; Casaus, J.; Cavallari, F.; Cavallo, N.; Cecchi, C.; Cerrada, M.; Chamizo, M.; Chang, Y.H.; Chemarin, M.; Chen, A.; Chen, G.; Chen, G.M.; Chen, H.F.; Chen, H.S.; Chiefari, G.; Cifarelli, L.; Cindolo, F.; Clare, I.; Clare, R.; Coignet, G.; Colino, N.; Costantini, S.; de la Cruz, B.; Cucciarelli, S.; van Dalen, J.A.; de Asmundis, R.; Deglon, P.; Debreczeni, J.; Degre, A.; Deiters, K.; della Volpe, D.; Delmeire, E.; Denes, P.; DeNotaristefani, F.; De Salvo, A.; Diemoz, M.; Dierckxsens, M.; Dionisi, C.; Dittmar, M.; Doria, A.; Dova, M.T.; Duchesneau, D.; Echenard, B.; Eline, A.; El Mamouni, H.; Engler, A.; Eppling, F.J.; Ewers, A.; Extermann, P.; Falagan, M.A.; Falciano, S.; Favara, A.; Fay, J.; Fedin, O.; Felcini, M.; Ferguson, T.; Fesefeldt, H.; Fiandrini, E.; Field, J.H.; Filthaut, F.; Fisher, P.H.; Fisher, W.; Fisk, I.; Forconi, G.; Freudenreich, K.; Furetta, C.; Galaktionov, Iouri; Ganguli, S.N.; Garcia-Abia, Pablo; Gataullin, M.; Gentile, S.; Giagu, S.; Gong, Z.F.; Grenier, Gerald Jean; Grimm, O.; Gruenewald, M.W.; Guida, M.; van Gulik, R.; Gupta, V.K.; Gurtu, A.; Gutay, L.J.; Haas, D.; Hakobyan, R.S.; Hatzifotiadou, D.; Hebbeker, T.; Herve, Alain; Hirschfelder, J.; Hofer, H.; Hohlmann, M.; Holzner, G.; Hou, S.R.; Hu, Y.; Jin, B.N.; Jones, Lawrence W.; de Jong, P.; Josa-Mutuberria, I.; Kafer, D.; Kaur, M.; Kienzle-Focacci, M.N.; Kim, J.K.; Kirkby, Jasper; Kittel, W.; Klimentov, A.; Konig, A.C.; Kopal, M.; Koutsenko, V.; Kraber, M.; Kraemer, R.W.; Krenz, W.; Kruger, A.; Kunin, A.; Ladron 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.; Levtchenko, P.; Li, C.; Likhoded, S.; Lin, C.H.; Lin, W.T.; Linde, F.L.; Lista, L.; Liu, Z.A.; Lohmann, W.; Longo, E.; Lu, Y.S.; Lubelsmeyer, K.; Luci, C.; Luminari, L.; Lustermann, W.; Ma, W.G.; Malgeri, L.; Malinin, A.; Mana, C.; Mangeol, D.; 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.; Kluge, Hannelies; Ofierzynski, R.; Organtini, G.; Palomares, C.; Pandoulas, D.; Paolucci, P.; Paramatti, R.; Passaleva, G.; Patricelli, S.; Paul, Thomas Cantzon; Pauluzzi, M.; Paus, C.; Pauss, F.; Pedace, M.; Pensotti, S.; Perret-Gallix, D.; Petersen, B.; Piccolo, D.; Pierella, F.; Pioppi, M.; Piroue, P.A.; Pistolesi, E.; Plyaskin, V.; Pohl, M.; Pojidaev, V.; Pothier, J.; Prokofev, D.O.; Prokofiev, D.; Quartieri, J.; Rahal-Callot, G.; Rahaman, M.A.; Raics, P.; Raja, N.; Ramelli, R.; Rancoita, P.G.; Ranieri, R.; Raspereza, A.; Razis, P.; Ren, D.; Rescigno, M.; Reucroft, S.; Riemann, S.; Riles, Keith; Roe, B.P.; Romero, L.; Rosca, A.; Rosier-Lees, S.; Roth, Stefan; Rosenbleck, C.; Roux, B.; Rubio, J.A.; Ruggiero, G.; Rykaczewski, H.; Sakharov, A.; Saremi, S.; Sarkar, S.; Salicio, J.; Sanchez, E.; Sanders, M.P.; Schafer, C.; Schegelsky, V.; Schmidt-Kaerst, S.; Schmitz, D.; Schopper, H.; Schotanus, D.J.; Schwering, G.; Sciacca, C.; Servoli, L.; Shevchenko, S.; Shivarov, N.; Shoutko, V.; Shumilov, E.; Shvorob, A.; Siedenburg, T.; Son, D.; Spillantini, P.; Steuer, M.; Stickland, D.P.; Stoyanov, B.; Straessner, A.; Sudhakar, K.; Sultanov, G.; Sun, L.Z.; Sushkov, S.; Suter, H.; Swain, J.D.; Szillasi, Z.; Tang, X.W.; Tarjan, P.; Tauscher, L.; Taylor, L.; Tellili, B.; Teyssier, D.; Timmermans, Charles; Ting, Samuel C.C.; Ting, S.M.; Tonwar, S.C.; Toth, J.; Tully, C.; Tung, K.L.; Ulbricht, J.; Valente, E.; Van de Walle, R.T.; Veszpremi, V.; Vesztergombi, G.; Vetlitsky, I.; Vicinanza, D.; Viertel, G.; Villa, S.; Vivargent, M.; Vlachos, S.; Vodopianov, 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.; Zoller, M.

    2002-01-01

    Strange baryon pair production in two-photon collisions is studied with the L3 detector at LEP. The analysis is based on data collected at e+e- centre-of-mass energies from 91 GeV to 208 GeV, corresponding to an integrated luminosity of 844 pb-1. The processes gamma gamma -> Lambda Anti-lambda and gamma gamma -> Sigma0 Anti-sigma0 are identified. Their cross sections as a function of the gamma gamma centre-of-mass energy are measured and results are compared to predictions of the quark-diquark model.

  2. Proton-Antiproton Pair Production in Two-Photon Collisions at LEP

    CERN Document Server

    Achard, P.; Aguilar-Benitez, M.; Alcaraz, J.; Alemanni, G.; Allaby, J.; Aloisio, A.; Alviggi, M.G.; Anderhub, H.; Andreev, Valery 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.; Barillere, R.; Bartalini, P.; Basile, M.; Batalova, N.; Battiston, R.; Bay, A.; Becattini, F.; Becker, U.; Behner, F.; Bellucci, L.; Berbeco, R.; Berdugo, J.; Berges, P.; Bertucci, B.; Betev, B.L.; Biasini, M.; Biglietti, M.; Biland, A.; Blaising, J.J.; Blyth, S.C.; Bobbink, G.J.; Bohm, A.; Boldizsar, L.; Borgia, B.; Bottai, S.; Bourilkov, D.; Bourquin, M.; Braccini, S.; Branson, J.G.; Brochu, F.; Burger, J.D.; Burger, W.J.; Cai, X.D.; Capell, M.; Cara Romeo, G.; Carlino, G.; Cartacci, A.; Casaus, J.; Cavallari, F.; Cavallo, N.; Cecchi, C.; Cerrada, M.; Chamizo, M.; Chang, Y.H.; Chemarin, M.; Chen, A.; Chen, G.; Chen, G.M.; Chen, H.F.; Chen, H.S.; Chiefari, G.; Cifarelli, L.; Cindolo, F.; Clare, I.; Clare, R.; Coignet, G.; Colino, N.; Costantini, S.; de la Cruz, B.; Cucciarelli, S.; van Dalen, J.A.; de Asmundis, R.; Deglon, P.; Debreczeni, J.; Degre, A.; Dehmelt, K.; Deiters, K.; della Volpe, D.; Delmeire, E.; Denes, P.; DeNotaristefani, F.; De Salvo, A.; Diemoz, M.; Dierckxsens, M.; Dionisi, C.; Dittmar, M.; Doria, A.; Dova, M.T.; Duchesneau, D.; Duda, M.; Echenard, B.; Eline, A.; El Hage, A.; El Mamouni, H.; Engler, A.; Eppling, F.J.; Extermann, P.; Falagan, M.A.; Falciano, S.; Favara, A.; Fay, J.; Fedin, O.; Felcini, M.; Ferguson, T.; Fesefeldt, H.; Fiandrini, E.; Field, J.H.; Filthaut, F.; Fisher, P.H.; Fisher, W.; Fisk, I.; Forconi, G.; Freudenreich, K.; Furetta, C.; Galaktionov, Iouri; Ganguli, S.N.; Garcia-Abia, Pablo; Gataullin, M.; Gentile, S.; Giagu, S.; Gong, Z.F.; Grenier, Gerald Jean; Grimm, O.; Gruenewald, M.W.; Guida, M.; van Gulik, R.; Gupta, V.K.; Gurtu, A.; Gutay, L.J.; Haas, D.; Hakobyan, R.S.; Hatzifotiadou, D.; Hebbeker, T.; Herve, Alain; Hirschfelder, J.; Hofer, H.; Hohlmann, M.; Holzner, G.; Hou, S.R.; Hu, Y.; Jin, B.N.; Jones, Lawrence W.; de Jong, P.; Josa-Mutuberria, I.; Kafer, D.; Kaur, M.; Kienzle-Focacci, M.N.; Kim, J.K.; Kirkby, Jasper; Kittel, W.; Klimentov, A.; Konig, A.C.; Kopal, M.; Koutsenko, V.; Kraber, M.; Kraemer, R.W.; Kruger, A.; Kunin, A.; Ladron 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.; Levtchenko, P.; Li, C.; Likhoded, S.; Lin, C.H.; Lin, W.T.; Linde, F.L.; Lista, L.; Liu, Z.A.; Lohmann, W.; Longo, E.; Lu, Y.S.; Luci, C.; Luminari, L.; Lustermann, W.; Ma, W.G.; Malgeri, L.; Malinin, A.; Mana, C.; Mans, J.; Martin, J.P.; Marzano, F.; Mazumdar, K.; McNeil, R.R.; Mele, S.; Merola, L.; Meschini, M.; Metzger, W.J.; Mihul, A.; Milcent, H.; Mirabelli, G.; Mnich, J.; Mohanty, G.B.; Muanza, G.S.; Muijs, A.J.M.; Musicar, B.; Musy, M.; Nagy, S.; Natale, S.; Napolitano, M.; Nessi-Tedaldi, F.; Newman, H.; Nisati, A.; Kluge, Hannelies; Ofierzynski, R.; Organtini, G.; Pal, I.; Palomares, C.; Paolucci, P.; Paramatti, R.; Passaleva, G.; Patricelli, S.; Paul, Thomas Cantzon; Pauluzzi, M.; Paus, C.; Pauss, F.; Pedace, M.; Pensotti, S.; Perret-Gallix, D.; Petersen, B.; Piccolo, D.; Pierella, F.; Pioppi, M.; Piroue, P.A.; Pistolesi, E.; Plyaskin, V.; Pohl, M.; Pojidaev, V.; Pothier, J.; Prokofev, D.; Quartieri, J.; Rahal-Callot, G.; Rahaman, Mohammad Azizur; Raics, P.; Raja, N.; Ramelli, R.; Rancoita, P.G.; Ranieri, R.; Raspereza, A.; Razis, P.; Ren, D.; Rescigno, M.; Reucroft, S.; Riemann, S.; Riles, Keith; Roe, B.P.; Romero, L.; Rosca, A.; Rosier-Lees, S.; Roth, Stefan; Rosenbleck, C.; Rubio, J.A.; Ruggiero, G.; Rykaczewski, H.; Sakharov, A.; Saremi, S.; Sarkar, S.; Salicio, J.; Sanchez, E.; Schafer, C.; Schegelsky, V.; Schopper, H.; Schotanus, D.J.; Sciacca, C.; Servoli, L.; Shevchenko, S.; Shivarov, N.; Shoutko, V.; Shumilov, E.; Shvorob, A.; Son, D.; Souga, C.; Spillantini, P.; Steuer, M.; Stickland, D.P.; Stoyanov, B.; Straessner, A.; Sudhakar, K.; Sultanov, G.; Sun, L.Z.; Sushkov, S.; Suter, H.; Swain, J.D.; Szillasi, Z.; Tang, X.W.; Tarjan, P.; Tauscher, L.; Taylor, L.; Tellili, B.; Teyssier, D.; Timmermans, Charles; Ting, Samuel C.C.; Ting, S.M.; Tonwar, S.C.; Toth, J.; Tully, C.; Tung, K.L.; Ulbricht, J.; Valente, E.; Van de Walle, R.T.; Vasquez, R.; Veszpremi, V.; Vesztergombi, G.; Vetlitsky, I.; Vicinanza, D.; Viertel, G.; Villa, S.; Vivargent, M.; Vlachos, S.; Vodopianov, I.; Vogel, H.; Vogt, H.; Vorobev, I.; Vorobyov, A.A.; Wadhwa, M.; Wang, Q.; Wang, X.L.; Wang, Z.M.; Weber, M.; Wienemann, P.; Wilkens, H.; Wynhoff, S.; Xia, L.; Xu, Z.Z.; Yamamoto, J.; Yang, B.Z.; Yang, C.G.; Yang, H.J.; Yang, M.; Yeh, S.C.; Zalite, An.; Zalite, Yu.; Zhang, Z.P.; Zhao, J.; Zhu, G.Y.; Zhu, R.Y.; Zhuang, H.L.; Zichichi, A.; Zimmermann, B.; Zoller, M.

    2003-01-01

    The reaction e+e- -> e+e- proton antiproton is studied with the L3 detector at LEP. The analysis is based on data collected at e+e- center-of-mass energies from 183 GeV to 209 GeV, corresponding to an integrated luminosity of 667 pb-1. The gamma gamma -> proton antiproton differential cross section is measured in the range of the two-photon center-of-mass energy from 2.1 GeV to 4.5 GeV. The results are compared to the predictions of the three-quark and quark-diquark models.

  3. Experimental preparation of two-photon Knill-Laflamme-Milburn states

    Science.gov (United States)

    Lemr, Karel; Černoch, Antonín; Soubusta, Jan; Fiurášek, Jaromír

    2010-01-01

    We report an experimental preparation of the so-called Knill-Laflamme-Milburn states (KLM states) that have been known to have interesting properties related to quantum information processing. Our experiment has demonstrated successful preparation of entangled two-photon four-mode KLM states using spontaneous parametric down-conversion as a source of entangled photon pairs, linear optical components, and avalanche photodiodes for single-photon detection. We have verified the successful generation of KLM states by complete quantum state tomography.

  4. Search for Anomalous Production of Events with Two Photons and Additional Energetic Objects at CDF

    Energy Technology Data Exchange (ETDEWEB)

    Aaltonen, T.; /Helsinki Inst. of Phys.; Adelman, J.; /Chicago U., EFI; Alvarez Gonzalez, B.; /Oviedo U. /Cantabria Inst. of Phys.; Amerio, S.; /Padua U. /INFN, Padua; Amidei, D.; /Michigan U.; Anastassov, A.; /Northwestern U.; Annovi, A.; /Frascati; Antos, J.; /Comenius U. /Kosice, IEF; Apollinari, G.; /Fermilab; Apresyan, A.; /Purdue U.; Arisawa, T.; /Waseda U. /Dubna, JINR

    2009-10-01

    The authors present results of a search for anomalous production of two photons together with an electron, muon, {tau} lepton, missing transverse energy, or jets using p{bar p} collision data from 1.1-2.0 fb{sup -1} of integrated luminosity collected by the Collider Detector at Fermilab (CDF). The event yields and kinematic distributions are examined for signs for new physics without favoring a specific model of new physics. The results are consistent with the standard model expectations. The search employs several new analysis techniques that significantly reduce instrumental backgrounds in channels with an electron and missing transverse energy.

  5. Two-photon production of leptons at hadron colliders in semielastic and elastic cases

    Energy Technology Data Exchange (ETDEWEB)

    Manko, A. Yu., E-mail: andrej.j.manko@gmail.com [National Academy of Sciences of Belarus, Institute of Physics (Belarus); Shulyakovsky, R. G., E-mail: shul@ifanbel.bas-net.by, E-mail: shulyakovsky@iaph.bas-net.by [National Academy of Sciences of Belarus, Institute of Applied Physics (Belarus)

    2016-03-15

    The mechanism of two-photon dilepton production is studied in the equivalent-photon (Weizsäcker–Williams) approximation. This approximation is shown to describe well experimental data from hadron accelerators. The respective total and differential cross sections were obtained for the LHC and for the Tevatron collider at various energies of colliding hadrons. The differential cross sections were studied versus the dilepton invariant mass, transverse momentum, and emission angle in the reference frame comoving with the center of mass of colliding hadrons. The cases of semielastic and inelastic collisions were examined.

  6. Exclusive production of pion and kaon meson pairs in two photon collisions at LEP

    CERN Document Server

    Heister, A; Antonelli, A; Antonelli, M; Armstrong, S R; Awunor, O; Azzurri, P; Badaud, F; Bagliesi, G; Barate, R; Barklow, Timothy L; Bencivenni, G; Berkelman, K; Beuselinck, R; Blair, G A; Bloch-Devaux, B; Blondel, A; Blumenschein, U; Boccali, T; Bonissent, A; Booth, C N; Borean, C; Bossi, F; Boucrot, J; Bouhova-Thacker, E; Boumediene, D E; Bowdery, C K; Brandt, S; Bravo, S; Brient, J C; Brunelière, R; Buchmüller, O L; Böhrer, A; Callot, O; Cameron, W; Capon, G; Cartwright, S; Casado, M P; Cattaneo, M; Cavanaugh, R J; Cerutti, F; Chiarella, V; Chmeissani, M; Ciulli, V; Clarke, D P; Clerbaux, B; Clifft, R W; Colaleo, A; Colas, P; Combley, F; Cowan, G; Coyle, P; Cranmer, K; Creanza, D; Crespo, J M; Curtil, C; David, A; Davier, M; Davies, G; De Bonis, I; De Filippis, N; De Palma, M; Delaere, C; Dessagne, S; Dhamotharan, S; Dietl, H; Dissertori, G; Dornan, P J; Drevermann, H; Duflot, L; Décamp, D; Ealet, A; Edgecock, T R; Ellis, G; Fabbro, B; Falvard, A; Fayolle, D; Ferguson, D P S; Fernández-Bosman, M; Fernández, E; Finch, A J; Focardi, E; Forty, R W; Foster, F; Fouchez, D; Foà, L; Frank, M; Ganis, G; Gao, Y; García-Bellido, A; Garrido, L; Gay, P; Geweniger, C; Ghete, V M; Giammanco, A; Giannini, G; Gianotti, F; Giassi, A; Girone, M; Girtler, P; González, S; Goy, C; Green, M G; Grivaz, J F; Grupen, C; Hanke, P; Hansen, J B; Hansen, J D; Hansen, J R; Hansen, P H; Harvey, J; Hayes, O J; He, H; Hepp, V; Hess, J; Heusse, P; Hill, R D; Hodgson, P N; Hu, H; Huang, X; Hughes, G; Hutchcroft, D E; Hölldorfer, F; Hüttmann, K; Iaselli, G; Jacholkowska, A; Jakobs, K; Janot, P; Jin, S; Jones, L T; Jones, R W L; Jost, B; Jousset, J; Jézéquel, S; Kado, M; Kayser, F; Kennedy, J; Kile, J; Kleinknecht, K; Kluge, E E; Kneringer, E; Kraan, A C; Kuhn, D; Kyriakis, A; Lançon, E; Laurelli, P; Lees, J P; Lehto, M H; Leibenguth, G; Lemaire, M C; Lemaître, V; Ligabue, F; Lin, J; Litke, A M; Locci, E; Lynch, J G; Lütjens, G; Machefert, F P; Maggi, G; Maggi, M; Mannocchi, G; Marinelli, N; Markou, C; Martin, F; Martínez, M; Mato, P; McNamara, P A; Medcalf, T; Merle, E; Messineo, A; Michel, B; Minard, M N; Misiejuk, A; Monteil, S; Moser, H G; Moutoussi, A; Murtas, G P; Männer, W; Müller, A S; Negus, P; Ngac, A; Nielsen, J; Nilsson, B S; Norton, P R; Nowell, J; Nuzzo, S; O'Shea, V; Ouyang, Q; Pacheco, A; Palla, Fabrizio; Pallin, D; Pan, Y B; Parrini, G; Pascolo, J M; Passalacqua, L; Payre, P; Pearson, M R; Perret, P; Pietrzyk, B; Prange, G; Putzer, A; Pérez, P; Pütz, J; Ragusa, F; Rander, J; Ranieri, A; Ranjard, F; Raso, G; Renk, B; Robertson, N A; Rolandi, Luigi; Rothberg, J E; Rougé, A; Rudolph, G; Ruggieri, F; Ruiz, H; Rutherford, S A; Sander, H G; Sanguinetti, G; Schael, S; Schlatter, W D; Schmeling, S; Sciabà, A; Sedgbeer, J K; Selvaggi, G; Serin, L; Settles, Ronald; Sguazzoni, G; Silvestris, L; Simopoulou, Errietta; Smizanska, M; Spagnolo, R; Stenzel, H; Strong, J A; Taylor, G; Teixeira-Dias, P; Tempesta, P; Tenchini, A; Teubert, F; Thompson, A S; Thompson, J C; Thompson, L F; Tilquin, A; Tittel, K; Tomalin, I R; Tricomi, A; Trocmé, B; Tuchming, B; Valassi, Andrea; Vallage, B; Vayaki, Anna; Veillet, J J; Venturi, P; Verdini, P G; Videau, H L; Videau, I; Villegas, M; Von Wimmersperg-Töller, J H; Wachsmuth, H W; Wang, T; Ward, J J; Wasserbaech, S R; White, R; Wiedenmann, W; Wolf, G; Wu, J; Wu Sau Lan; Wu, X; Wunsch, M; Xie, Y; Xu, R; Xue, S; Zachariadou, K; Zeitnitz, C; Zhang, J; Zhang, L; Zhao, W; Ziegler, T; Zito, G; Zobernig, G; van der Aa, O

    2003-01-01

    Exclusive production of $\\pi$ and K meson pairs in two photon collisions is measured with ALEPH data collected between 1992 and 2000. Cross sections are presented as a function of \\cos\\theta^* and invariant mass, for |\\cos\\theta^* |< 0.6 and invariant masses between 2.0 and 6.0 \\mathrm{GeV}/c^2 (2.25 and 4.0 \\mathrm{GeV}/c^2) for pions (kaons). The shape of the distributions are found to be well described by QCD predictions but the data have a significantly higher normalisation.

  7. Polarizable Embedded RI-CC2 Method for Two-Photon Absorption Calculations

    DEFF Research Database (Denmark)

    Hršak, Dalibor; Khah, Alireza Marefat; Christiansen, Ove

    2015-01-01

    We present a novel polarizable embedded resolution-of-identity coupled cluster singles and approximate doubles (PERI-CC2) method for calculation of two-photon absorption (TPA) spectra of large molecular systems. The method was benchmarked for three types of systems: a water-solvated molecule...... of formamide, a uracil molecule in aqueous solution, and a set of mutants of the channelrhodopsin (ChR) protein. The first test case shows that the PERI-CC2 method is in excellent agreement with the PE-CC2 method and in good agreement with the PE-CCSD method. The uracil test case indicates that the effects...

  8. Two-photon excitation/ionization of the 1s-shell of the argon atom

    CERN Document Server

    Novikov, S A

    2002-01-01

    The absolute values and the shape of the two-photon excitation/ionization cross section of the 1s-shell of the argon atom are calculated with inclusion of the many-particle effects, i.e., the relaxation of the atomic residue in the field of the vacancies created, and the decay of the vacancies into the channels of Auger and (or) radiative types. The wavefunctions of the one-particle states are calculated in non-relativistic approximation. The calculations are performed for both linear and circular polarization of the laser beam.

  9. Two photon laser spectroscopy of antiprotonic helium atoms at CERN’s AD

    CERN Document Server

    Hori, M

    2014-01-01

    The ASACUSA collaboration of CERN has carried out two-photon laser spectroscopy of antiprotonic helium atoms using counter-propagating ultraviolet laser beams. This excited some non-linear transitions of the antiproton at the wavelengths λ = 139.8–197.0 nm, in a way that reduced the thermal Doppler broadening of the observed resonances. The resulting narrow spectral lines allowed the measurement of three transition frequencies with fractional precisions of 2.3–5 parts in 109. By comparing these values with three-body QED calculations, the antiproton-to-electron mass ratio was derived as 1836.1526736(23). We briefly review these results.

  10. Electron angular distributions in two-photon double ionization of helium

    Energy Technology Data Exchange (ETDEWEB)

    Makris, M.G. [Univ. of Crete, Physics Dept., Heraklion (Greece); Nikolopoulos, L.A.A. [Inst. of Electronic Structure and Laser, Forth, Heraklion, Crete (Greece); Lambropoulos, P. [Univ. of Crete, Physics Dept., Heraklion (Greece); Inst. of Electronic Structure and Laser, Forth, Heraklion, Crete (Greece)

    2001-06-15

    We present photoelectron angular distributions (PAD) in two-photon double ionization of helium for photon energies in the range of 45 eV, where the direct double ejection is unequivocally distinguishable from the sequential. Through an approach that allows us to explore the role of correlation, we obtain conditions under which the two electrons have the tendency to be emitted in the same direction. Relevant experiments should be feasible either using high-order harmonic generation or the upcoming short-wavelength free electron laser sources. (orig.)0.

  11. Tunable two-photon correlation in a double-cavity optomechanical system

    Directory of Open Access Journals (Sweden)

    Zhi-Bo Feng

    2015-12-01

    Full Text Available Correlated photons are essential sources for quantum information processing. We propose a practical scheme to generate pairs of correlated photons in a controllable fashion from a double-cavity optomechanical system, where the variable optomechanical coupling strength makes it possible to tune the photon correlation at our will. The key operation is based on the repulsive or attractive interaction between the two photons intermediated by the mechanical resonator. The present protocol could provide a potential approach to coherent control of the photon correlation using the optomechanical cavity.

  12. Forward two-photon exchange in elastic lepton-proton scattering and hyperfine-splitting correction

    Energy Technology Data Exchange (ETDEWEB)

    Tomalak, Oleksandr [Johannes Gutenberg Universitaet, Institut fuer Kernphysik and PRISMA Cluster of Excellence, Mainz (Germany)

    2017-08-15

    We relate the forward two-photon exchange (TPE) amplitudes to integrals of the inclusive lepton-proton scattering cross sections. These relations yield an alternative way for the evaluation of the TPE correction to hyperfine-splitting (HFS) in the hydrogen-like atoms with an equivalent to the standard approach (Iddings, Drell and Sullivan) result implying the Burkhardt-Cottingham sum rule. For evaluation of the individual effects (e.g., elastic contribution) our approach yields a distinct result. We compare both methods numerically on examples of the elastic contribution and the full TPE correction to HFS in electronic and muonic hydrogen. (orig.)

  13. Search for Standard Model Higgs boson in the two-photon final state in ATLAS

    Directory of Open Access Journals (Sweden)

    Davignon Olivier

    2012-06-01

    Full Text Available We report on the search for the Standard Model Higgs boson decaying into two photons based on proton-proton collision data with a center-of-mass energy of 7 TeV recorded by the ATLAS experiment at the LHC. The dataset has an integrated luminosity of about 1:08 fb−1. The expected cross section exclusion at 95% confidence level varies between 2:0 and 5:8 times the Standard Model cross section over the diphoton mass range 110 – 150 GeV. The maximum deviations from the background-only expectation are consistent with statistical fluctuations.

  14. Autocorrelation and phase retrieval in the UV using two-photon absorption in diamond pin photodiodes.

    Science.gov (United States)

    Kleimeier, Nils Fabian; Haarlammert, Thorben; Witte, Henrik; Schühle, Udo; Hochedez, Jean-Francois; BenMoussa, Ali; Zacharias, Helmut

    2010-03-29

    We report on the utilization of the two-photon induced free carrier generation in a diamond pin-type photodiode to record fringe-resolved second-order autocorrelations of femtosecond pulses in the UV. Measurements in photovoltaic mode are performed at the second and third harmonic of a Ti:sapphire laser (lambda(0)=401 nm and lambda(0)=265 nm) with pulse energies down to about 2 nJ. The band gap of diamond of 5.5 eV sets a short wavelength limit at about 225 nm. Combined with the simultaneously recorded linear autocorrelation the spectral phase is reconstructed employing an iterative algorithm.

  15. Two-Photon Ionization of He through a Superposition of Higher Harmonics.

    Science.gov (United States)

    Papadogiannis, N A; Nikolopoulos, L A A; Charalambidis, D; Tsakiris, G D; Tzallas, P; Witte, K

    2003-04-04

    We present experimental results and theoretical analysis of two-photon ionization of He by a superposition of the 7th to the 13th harmonic of a Ti:sapphire laser. Solving the time-dependent Schrödinger equation for He in a coherent polychromatic field, the He+ yield is calculated. From this yield the number of He+ ions produced has been estimated and found in reasonable agreement with its measured value. The present results establish the feasibility of a second-order autocorrelation measurement of superposition of harmonics, and thus they represent the precursor towards the direct temporal characterization of attosecond pulse trains.

  16. General calculation of the cross section for dark matter annihilations into two photons

    Science.gov (United States)

    Garcia-Cely, Camilo; Rivera, Andres

    2017-03-01

    Assuming that the underlying model satisfies some general requirements such as renormalizability and CP conservation, we calculate the non-relativistic one-loop cross section for any self-conjugate dark matter particle annihilating into two photons. We accomplish this by carefully classifying all possible one-loop diagrams and, from them, reading off the dark matter interactions with the particles running in the loop. Our approach is general and leads to the same results found in the literature for popular dark matter candidates such as the neutralinos of the MSSM, minimal dark matter, inert Higgs and Kaluza-Klein dark matter.

  17. TWO-PHOTON PHYSICS IN NUCLEUS-NUCLEUS COLLISIONS AT RHIC.

    Energy Technology Data Exchange (ETDEWEB)

    NYSTRAND,J.

    1998-09-10

    Ultra-relativistic heavy-ions carry strong electromagnetic and nuclear fields. Interactions between these fields in peripheral nucleus-nucleus collisions can probe many interesting physics topics. This presentation will focus on coherent two-photon and photonuclear processes at RHIC. The rates for these interactions will be high. The coherent coupling of all the protons in the nucleus enhances the equivalent photon flux by a factor Z{sup 2} up to an energy of {approx} 3 GeV. The plans for studying coherent interactions with the STAR experiment will be discussed. Experimental techniques for separating signal from background will be presented.

  18. Two-photon physics in nucleus-nucleus collisions at RHIC

    Energy Technology Data Exchange (ETDEWEB)

    Nystrand, J.; Klein, S.

    1998-09-01

    Ultra-relativistic heavy-ions carry strong electromagnetic and nuclear fields. Interactions between these fields in peripheral nucleus-nucleus collisions can probe many interesting physics topics. This presentation will focus on coherent two-photon and photonuclear processes at RHIC. The rates for these interactions will be high. The coherent coupling of all the protons in the nucleus enhances the equivalent photon flux by a factor Z{sup 2} up to an energy of {approx} 3 GeV. The plans for studying coherent interactions with the STAR experiment will be discussed. Experimental techniques for separating signal from background will be presented.

  19. Strategies for rapid and reliable fabrication of microoptical structures using two-photon polymerization

    Science.gov (United States)

    Steenhusen, Sönke; Hasselmann, Sebastian; Domann, Gerhard

    2017-02-01

    Two-Photon Polymerization (2PP) has attracted broad interest for the fabrication of microoptical elements due to its design flexibility and precision. Along with tailored hybrid polymers a higher level of functional integration and new application concepts are enabled. As the entire volume of the desired 3D structure is filled in a point-by-point fashion, the fabrication can require several days inhibiting the adoption of 2PP as an additive manufacturing process at industrial level. We review different strategies to overcome the limitation in throughput and their impact on the patterning result. Particularly, processing using galvoscanner technology and replication of 2PP structures are highlighted.

  20. Suitable photo-resists for two-photon polymerization using femtosecond fiber lasers

    KAUST Repository

    Rajamanickam, V.P.

    2014-06-01

    We present suitable materials with good optical and mechanical properties, simple processing, efficient and optimized for two-photon polymerization (TPP) with femtosecond fiber lasers. We selected readily available acrylic monomer Bisphenol A ethoxylate diacrylate (BPA-EDA) with three different photo-initiators (PIs), isopropyl thioxanthone (ITX), 7-diethylamino-3-thenoylcoumarin (DETC), and 4,4′ bis(diethylamino) benzophenone (BDEB), since their absorption spectra match well with the laser wavelength at 780 nm. These PIs grant efficient radical generation, reactivity and high solubility in acrylic monomers. Finally, good optical and mechanical properties are demonstrated by the fabrication of different micro-structures.

  1. Novel xenon calibration scheme for two-photon absorption laser induced fluorescence of hydrogen

    Energy Technology Data Exchange (ETDEWEB)

    Elliott, Drew; Scime, Earl; Short, Zachary, E-mail: zdshort@mix.wvu.edu [Department of Physics and Astronomy, West Virginia University, Morgantown, West Virginia 26056 (United States)

    2016-11-15

    Two photon absorption laser induced fluorescence (TALIF) measurements of neutral hydrogen and its isotopes are typically calibrated by performing TALIF measurements on krypton with the same diagnostic system and using the known ratio of the absorption cross sections [K. Niemi et al., J. Phys. D 34, 2330 (2001)]. Here we present the measurements of a new calibration method based on a ground state xenon scheme for which the fluorescent emission wavelength is nearly identical to that of hydrogen, thereby eliminating chromatic effects in the collection optics and simplifying detector calibration. We determine that the ratio of the TALIF cross sections of xenon and hydrogen is 0.024 ± 0.001.

  2. A stable frequency comb directly referenced to rubidium electromagnetically induced transparency and two-photon transitions

    Energy Technology Data Exchange (ETDEWEB)

    Hou, Dong; Wu, Jiutao; Zhang, Shuangyou; Ren, Quansheng; Zhang, Zhigang; Zhao, Jianye, E-mail: zhaojianye@pku.edu.cn [Department of Electronics, Peking University, Beijing, 100871 (China)

    2014-03-17

    We demonstrate an approach to create a stable erbium-fiber-based frequency comb at communication band by directly locking the combs to two rubidium atomic transitions resonances (electromagnetically induced transparency absorption and two-photon absorption), respectively. This approach directly transfers the precision and stability of the atomic transitions to the comb. With its distinguishing feature of compactness by removing the conventional octave-spanning spectrum and f-to-2f beating facilities and the ability to directly control the comb's frequency at the atomic transition frequency, this stable optical comb can be widely used in optical communication, frequency standard, and optical spectroscopy and microscopy.

  3. Two-photon polymerization of cylinder microstructures by femtosecond Bessel beams

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Liang [Department of Precision Machinery and Precision Instrumentation, University of Science and Technology of China, Hefei 230026 (China); Laser Zentrum Hannover e.V., 30419 Hannover (Germany); El-Tamer, Ayman; Hinze, Ulf; Chichkov, Boris N [Laser Zentrum Hannover e.V., 30419 Hannover (Germany); Li, Jiawen, E-mail: jwl@ustc.edu.cn; Hu, Yanlei; Huang, Wenhao; Chu, Jiaru [Department of Precision Machinery and Precision Instrumentation, University of Science and Technology of China, Hefei 230026 (China)

    2014-07-28

    In this work, we present an approach to modulate femtosecond laser beams into Bessel beams with a spatial light modulator (SLM) for two-photon polymerization applications. Bessel beams with different parameters are generated and annular optical fields are produced at the focal plane of the objective. Uniform cylinder microstructures are fabricated by a single illumination during a few seconds without stage translation. By modulating the holograms encoded on the SLM, the diameters of the fabricated annular structures can be flexibly controlled in a wide range with no need of changing the optical elements and realignment of the optical path.

  4. K$^{0}$($\\overline{K}^{0}$) production in two-photon processes at TRISTAN

    CERN Document Server

    Enomoto, R; Abe, T; Adachi, I; Adachi, K; Aoki, M; Awa, S; Emi, K; Fujii, H; Fujii, K; Fujii, T; Fujimoto, J; Fujita, K; Fujiwara, N; Hayashii, H; Howell, B; Iida, N; Itoh, R; Inoue, Y; Iwasaki, H; Iwasaki, M; Kaneyuki, K; Kajikawa, R; Kato, S; Kawabata, S; Kichimi, H; Kobayashi, M; Koltick, D S; Levine, I; Minami, S; Miyabayashi, K; Miyamoto, A; Muramatsu, K; Nagai, K; Nakabayashi, K; Nakano, E; Nitoh, O; Noguchi, S; Ochi, A; Ochiai, F; Ohishi, N; Ohnishi, Y; Ohshima, Y; Okuno, H; Okusawa, T; Shinohara, T; Sugiyama, A; Suzuki, S; Takahashi, K; Takahashi, T; Tanimori, T; Tauchi, T; Teramoto, Y; Toomi, N; Tsukamoto, T; Tsumura, O; Uno, S; Watanabe, T; Watanabe, Y; Yamaguchi, A; Yamamoto, A; Yamauchi, M

    1994-01-01

    We have carried out an inclusive measurement of K^0(\\overline{K^0}) production in two-photon processes at TRISTAN. The mean \\sqrt{s} was 58 GeV and the integrated luminosity was 199 pb^{-1}. High-statistics K_s samples were obtained under such conditions as no-, anti-electron, and remnant-jet tags. The remnant-jet tag, in particular, allowed us, for the first time, to measure the cross sections separately for the resolved-photon and direct processes.

  5. Observation of excess $\\lambda\\overline{\\lambda}$ production in two-photon processes at TRISTAN

    CERN Document Server

    Enomoto, R; Abe, T; Adachi, I; Adachi, K; Aoki, M; Awa, S; Emi, K; Fujii, H; Fujii, K; Fujii, T; Fujimoto, J; Fujita, K; Fujiwara, N; Hayashii, H; Howell, B; Iida, N; Itoh, R; Inoue, Y; Iwasaki, H; Iwasaki, M; Kaneyuki, K; Kajikawa, R; Kato, S; Kawabata, S; Kichimi, H; Kobayashi, M; Koltick, D S; Levine, I; Minami, S; Miyabayashi, K; Miyamoto, A; Muramatsu, K; Nagai, K; Nakabayashi, K; Nakano, E; Nitoh, O; Noguchi, S; Ochi, A; Ochiai, F; Ohishi, N; Ohnishi, Y; Ohshima, Y; Okuno, H; Okusawa, T; Shinohara, T; Sugiyama, A; Suzuki, S; Takahashi, K; Takahashi, T; Tanimori, T; Tauchi, T; Teramoto, Y; Toomi, N; Tsukamoto, T; Tsumura, O; Uno, S; Watanabe, T; Watanabe, Y; Yamaguchi, A; Yamamoto, A; Yamauchi, M; Enomoto, R

    1995-01-01

    We have carried out inclusive measurements of \\Lambda(\\overline{\\Lamb da}) production in two-photon processes at TRISTAN. The mean \\sqrt{s} was 58 GeV and the integrated luminosity was 265 pb^{-1}. Inclusive \\Lambda (\\overline{\\Lambda}) samples were obtained under such conditions as no-electron, anti-electron, and remnant-jet tags. The data were compared with theoretical calculations. The measured cross sections are two-times larger than the leading-order theoretical predictions, suggesting the necessity of next-to-leading-order Monte-Carlo generator.

  6. Two-photon polymerization for production of human iPSC-derived retinal cell grafts.

    Science.gov (United States)

    Worthington, Kristan S; Wiley, Luke A; Kaalberg, Emily E; Collins, Malia M; Mullins, Robert F; Stone, Edwin M; Tucker, Budd A

    2017-06-01

    Recent advances in induced pluripotent stem cell (iPSC) technology have paved the way for the production of patient-specific neurons that are ideal for autologous cell replacement for treatment of neurodegenerative diseases. In the case of retinal degeneration and associated photoreceptor cell therapy, polymer scaffolds are critical for cellular survival and integration; however, prior attempts to materialize this concept have been unsuccessful in part due to the materials' inability to guide cell alignment. In this work, we used two-photon polymerization to create 180μm wide non-degradable prototype photoreceptor scaffolds with varying pore sizes, slicing distances, hatching distances and hatching types. Hatching distance and hatching type were significant factors for the error of vertical pore diameter, while slicing distance and hatching type most affected the integrity and geometry of horizontal pores. We optimized printing parameters in terms of structural integrity and printing time in order to create 1mm wide scaffolds for cell loading studies. We fabricated these larger structures directly on a porous membrane with 3µm diameter pores and seeded them with human iPSC-derived retinal progenitor cells. After two days in culture, cells nested in and extended neuronal processes parallel to the vertical pores of the scaffolds, with maximum cell loading occurring in 25μm diameter pores. These results highlight the feasibility of using this technique as part of an autologous stem cell strategy for restoring vision to patients affected with retinal degenerative diseases. Cell replacement therapy is an important goal for investigators aiming to restore neural function to those suffering from neurodegenerative disease. Cell delivery scaffolds are frequently necessary for the success of such treatments, but traditional biomaterials often fail to facilitate the neuronal orientation and close packing needed to recapitulate the in vivo environment. Here, we use two-photon

  7. Giant Two-photon Absorption in Circular Graphene Quantum Dots in Infrared Region

    Science.gov (United States)

    Feng, Xiaobo; Li, Zhisong; Li, Xin; Liu, Yingkai

    2016-01-01

    We investigate theoretically the two-photon absorption (TPA) for circular graphene quantum dots (GQDs) with the edge of armchair and zigzag on the basis of electronic energy states obtained by solving the Dirac-Weyl equation numerically under finite difference method. The expressions for TPA cross section are derived and the transition selection rules are obtained. Results reveal that the TPA is significantly greater in GQDs than conventional semiconductor QDs in infrared spectrum (2–6 um) with a resonant TPA cross section of up to 1011 GM. The TPA peaks are tuned by the GQDs’ size, edge and electron relaxation rate. PMID:27629800

  8. Exploring the interactions between peptides and lipid bilayers using coherent anti-Stokes Raman scattering and two-photon fluorescence

    Science.gov (United States)

    Mari, M.; Mouras, R.; Downes, A.; Elfick, A.

    2011-06-01

    We have used a versatile and powerful microscope[1] for multi-modal biomedical imaging on which we combine Coherent Anti-Stokes Raman Scattering (CARS) with Two Photon Excitation Fluorescence (TPEF) using a Nd: YVO4 pump laser. We acquired 2PEF, CARS, and phase contrast images of Multilamellar Vesicles (MLVs) and Giant Unilamellar Vesicles (GUVs), as well as Raman spectra of the constituent lipids. A wide range of peptides are harmful to cells by altering the structure of the biological membranes. This effect depends on the composition of the membrane and the chemical structure of the peptide. The peptide we studied is the beta amyloid Aβ which is a major component of the amyloid plaques deposited on neuronal membranes of Alzheimer's disease (AD) patients. AD is neurodegenerative disorder in which the hallmark symptoms include cognitive decline and dementia[2] and is characterized by the formation of extracellular amyloid fibrils on the neuronal membranes of the brain. Many questions still remain unanswered concerning the destabilization of cellular ionic homeostasis due to pores formed during the interactions of lipid membranes with peptides. In this project, biomimics of cell membranes are used. The structures that best mimic the plasma membranes are MLVs or GUVs. These vesicles are formed using the gentle hydration technique[3] or the electroformation technique[4] respectively and are composed of phospholipids such as DOPC, DPPC, D62PPC and their binary mixtures. The MLVs and GUVs imaging by CARS and TPEF microscopy not only permits the direct imaging of the leakage phenomenon caused by the toxic peptide (Aβ) on the lipid bilayer, but also records simultaneously the lateral structure of the bilayer and peptide distribution in the plane across the membrane.

  9. Investigation on the SEL Sensitive Depth of an SRAM Using Linear and Two-Photon Absorption Laser Testing

    Science.gov (United States)

    Faraud, E.; Pouget, V.; Shao, K.; Larue, C.; Darracq, F.; Lewis, D.; Samaras, A.; Bezerra, F.; Lorfevre, E.; Ecoffet, R.

    2011-12-01

    Linear and two-photon laser testing is used to investigate the single-event latchup sensitive depth of SRAM CY7C1069 embedded in CARMEN satellite experiment. Results are discussed and compared with heavy ion and flight data.

  10. Single-shot two-photon exposure of commercial photoresist for the production of three-dimensional structures.

    Science.gov (United States)

    Witzgall, G; Vrijen, R; Yablonovitch, E; Doan, V; Schwartz, B J

    1998-11-15

    We report the use of an amplified femtosecond laser for single-shot two-photon exposure of the commercial photoresist SU-8. By scanning of the focal volume through the interior of the resist, three-dimensional (3-D) structures are fabricated on a shot-by-shot basis. The 800-nm two-photon exposure and damage thresholds are 3.2 and 8.1TW/cm(2), respectively. The nonlinear nature of the two-photon process allows the production of features that are smaller than the diffraction limit. Preliminary results suggest that Ti:sapphire oscillators can achieve single-shot two-photon exposure with thresholds as low as 1.6TW/cm(2) at 700 nm, allowing 3-D structures to be constructed at megahertz repetition rates.

  11. ARTICLE Theoretical Studies on One-photon and Two-photon Absorption Properties of Pyrene-core Derivatives

    Science.gov (United States)

    Ding, Ming-cui; Zhang, Zhen; Song, Yang; Wang, Chuan-kui

    2010-12-01

    The analytic response theory at density functional theory level is applied to investigate one-photon and two-photon absorption properties of a series of recently synthesized pyrene-core derivatives. The theoretical results show that there are a few charge-transfer states for each compound in the lower energy region. The one-photon absorption properties of the five investigated compounds are highly consistent with those given by experimental measurements. The two-photon absorption intensities of the compounds are greatly enhanced with the increments of the molecular sizes, in which the two-photon absorption cross section of the four-branched compound is about 5.6 times of that of the mono-branched molecule. Furthermore, it is shown that the two-photon absorption properties are sensitive to the geometrical arrangements.

  12. Two-photon polymerization of 3-D zirconium oxide hybrid scaffolds for long-term stem cell growth.

    Science.gov (United States)

    Skoog, Shelby A; Nguyen, Alexander K; Kumar, Girish; Zheng, Jiwen; Goering, Peter L; Koroleva, Anastasia; Chichkov, Boris N; Narayan, Roger J

    2014-06-01

    Two-photon polymerization is a technique that involves simultaneous absorption of two photons from a femtosecond laser for selective polymerization of a photosensitive material. In this study, two-photon polymerization was used for layer-by-layer fabrication of 3-D scaffolds composed of an inorganic-organic zirconium oxide hybrid material. Four types of scaffold microarchitectures were created, which exhibit layers of parallel line features at various orientations as well as pores between the line features. Long-term cell culture studies involving human bone marrow stromal cells were conducted using these 3-D scaffolds. Cellular adhesion and proliferation were demonstrated on all of the scaffold types; tissuelike structure was shown to span the pores. This study indicates that two-photon polymerization may be used to create microstructured scaffolds out of an inorganic-organic zirconium oxide hybrid material for use in 3-D tissue culture systems.

  13. Search for New Physics with Two Photons in the Final State with the ATLAS Detector

    CERN Document Server

    AUTHOR|(INSPIRE)INSPIRE-00441752

    This thesis reports on the search for new physics in the diphoton decay channel with the proton-proton collision data collected by ATLAS at a centre-of-mass energy of $\\sqrt{s}=8$~TeV in 2012 and $\\sqrt{s}=13$~TeV in 2015 and 2016. A feasibility study of the search for a pseudoscalar $A$ decaying to a $Z$ boson and a 125~GeV Higgs boson in the context of an extended Higgs sector, namedly the two-Higgs-doublet models, is presented. The search is performed with a final state of two jets and two photons using 20.3~${\\rm fb}^{-1}$ of data at $\\sqrt{s}=8$~TeV. The expected sensitivity is found to be competitive with the analysis with a final state of two electrons or muons and two $\\tau$ leptons, but less sensitive to the other searches with the Higgs decaying to a pair of $b$-quarks. Search for high mass resonances decaying to two photons at $\\sqrt{s}=13$~TeV is also presented. The analysed dataset corresponds to an integrated luminosity of $3.2~{\\rm fb}^{-1}$ in 2015 and $12.2~{\\rm fb}^{-1}$ in 2016. Two searche...

  14. In situ electrical and thermal monitoring of printed electronics by two-photon mapping.

    Science.gov (United States)

    Pastorelli, Francesco; Accanto, Nicolò; Jørgensen, Mikkel; van Hulst, Niek F; Krebs, Frederik C

    2017-06-19

    Printed electronics is emerging as a new, large scale and cost effective technology that will be disruptive in fields such as energy harvesting, consumer electronics and medical sensors. The performance of printed electronic devices relies principally on the carrier mobility and molecular packing of the polymer semiconductor material. Unfortunately, the analysis of such materials is generally performed with destructive techniques, which are hard to make compatible with in situ measurements, and pose a great obstacle for the mass production of printed electronics devices. A rapid, in situ, non-destructive and low-cost testing method is needed. In this study, we demonstrate that nonlinear optical microscopy is a promising technique to achieve this goal. Using ultrashort laser pulses we stimulate two-photon absorption in a roll coated polymer semiconductor and map the resulting two-photon induced photoluminescence and second harmonic response. We show that, in our experimental conditions, it is possible to relate the total amount of photoluminescence detected to important material properties such as the charge carrier density and the molecular packing of the printed polymer material, all with a spatial resolution of 400 nm. Importantly, this technique can be extended to the real time mapping of the polymer semiconductor film, even during the printing process, in which the high printing speed poses the need for equally high acquisition rates.

  15. Evidence for the direct two-photon transition from ψ(3686) to J/ψ.

    Science.gov (United States)

    Ablikim, M; Achasov, M N; Ambrose, D J; An, F F; An, Q; An, Z H; Bai, J Z; Ferroli, R B; Ban, Y; Becker, J; Berger, N; Bertani, M B; Bian, J M; Boger, E; Bondarenko, O; Boyko, I; Briere, R A; Bytev, V; Cai, X; Calcaterra, A C; Cao, G F; Chang, J F; Chelkov, G; Chen, G; Chen, H S; Chen, J C; Chen, M L; Chen, S J; Chen, Y; Chen, Y B; Cheng, H P; Chu, Y P; Cronin-Hennessy, D; Dai, H L; Dai, J P; Dedovich, D; Deng, Z Y; Denig, A; Denysenko, I; Destefanis, M; Ding, W M; Ding, Y; Dong, L Y; Dong, M Y; Du, S X; Fang, J; Fang, S S; Fava, L; Feldbauer, F; Feng, C Q; Fu, C D; Fu, J L; Gao, Y; Geng, C; Goetzen, K; Gong, W X; Gradl, W; Greco, M; Gu, M H; Gu, Y T; Guan, Y H; Guo, A Q; Guo, L B; Guo, Y P; Han, Y L; Hao, X Q; Harris, F A; He, K L; He, M; He, Z Y; Held, T; Heng, Y K; Hou, Z L; Hu, H M; Hu, J F; Hu, T; Huang, B; Huang, G M; Huang, J S; Huang, X T; Huang, Y P; Hussain, T; Ji, C S; Ji, Q; Ji, X B; Ji, X L; Jia, L K; Jiang, L L; Jiang, X S; Jiao, J B; Jiao, Z; Jin, D P; Jin, S; Jing, F F; Kalantar-Nayestanaki, N; Kavatsyuk, M; Kuehn, W; Lai, W; Lange, J S; Leung, J K C; Li, C H; Li, Cheng; Li, Cui; Li, D M; Li, F; Li, G; Li, H B; Li, J C; Li, K; Li, Lei; Li, N B; Li, Q J; Li, S L; Li, W D; Li, W G; Li, X L; Li, X N; Li, X Q; Li, X R; Li, Z B; Liang, H; Liang, Y F; Liang, Y T; Liao, G R; Liao, X T; Liu, B J; Liu, B J; Liu, C L; Liu, C X; Liu, C Y; Liu, F H; Liu, Fang; Liu, Feng; Liu, H; Liu, H B; Liu, H H; Liu, H M; Liu, H W; Liu, J P; Liu, Kun; Liu, Kai; Liu, K Y; Liu, P L; Liu, S B; Liu, X; Liu, X H; Liu, Y B; Liu, Y; Liu, Z A; Liu, Zhiqiang; Liu, Zhiqing; Loehner, H; Lu, G R; Lu, H J; Lu, J G; Lu, Q W; Lu, X R; Lu, Y P; Luo, C L; Luo, M X; Luo, T; Luo, X L; Lv, M; Ma, C L; Ma, F C; Ma, H L; Ma, Q M; Ma, S; Ma, T; Ma, X Y; Ma, Y; Maas, F E; Maggiora, M; Malik, Q A; Mao, H; Mao, Y J; Mao, Z P; Messchendorp, J G; Min, J; Min, T J; Mitchell, R E; Mo, X H; Morales Morales, C; Motzko, C; Muchnoi, N Yu; Nefedov, Y; Nicholson, C; Nikolaev, I B; Ning, Z; Olsen, S L; Ouyang, Q; Pacetti, S P; Park, J W; Pelizaeus, M; Peters, K; Ping, J L; Ping, R G; Poling, R; Prencipe, E; Pun, C S J; Qi, M; Qian, S; Qiao, C F; Qin, X S; Qin, Y; Qin, Z H; Qiu, J F; Rashid, K H; Rong, G; Ruan, X D; Sarantsev, A; Schulze, J; Shao, M; Shen, C P; Shen, X Y; Sheng, H Y; Shepherd, M R; Song, X Y; Spataro, S; Spruck, B; Sun, D H; Sun, G X; Sun, J F; Sun, S S; Sun, X D; Sun, Y J; Sun, Y Z; Sun, Z J; Sun, Z T; Tang, C J; Tang, X; Thorndike, E H; Tian, H L; Toth, D; Ulrich, M U; Varner, G S; Wang, B; Wang, B Q; Wang, K; Wang, L L; Wang, L S; Wang, M; Wang, P; Wang, P L; Wang, Q; Wang, Q J; Wang, S G; Wang, X F; Wang, X L; Wang, Y D; Wang, Y F; Wang, Y Q; Wang, Z; Wang, Z G; Wang, Z Y; Wei, D H; Weidenkaff, P; Wen, Q G; Wen, S P; Werner, M W; Wiedner, U; Wu, L H; Wu, N; Wu, S X; Wu, W; Wu, Z; Xia, L G; Xiao, Z J; Xie, Y G; Xiu, Q L; Xu, G F; Xu, G M; Xu, H; Xu, Q J; Xu, X P; Xu, Y; Xu, Z R; Xue, F; Xue, Z; Yan, L; Yan, W B; Yan, Y H; Yang, H X; Yang, T; Yang, Y; Yang, Y X; Ye, H; Ye, M; Ye, M H; Yu, B X; Yu, C X; Yu, J S; Yu, S P; Yuan, C Z; Yuan, W L; Yuan, Y; Zafar, A A; Zallo, A Z; Zeng, Y; Zhang, B X; Zhang, B Y; Zhang, C C; Zhang, D H; Zhang, H H; Zhang, H Y; Zhang, J; Zhang, J G; Zhang, J Q; Zhang, J W; Zhang, J Y; Zhang, J Z; Zhang, L; Zhang, S H; Zhang, T R; Zhang, X J; Zhang, X Y; Zhang, Y; Zhang, Y H; Zhang, Y S; Zhang, Z P; Zhang, Z Y; Zhao, G; Zhao, H S; Zhao, J W; Zhao, K X; Zhao, Lei; Zhao, Ling; Zhao, M G; Zhao, Q; Zhao, S J; Zhao, T C; Zhao, X H; Zhao, Y B; Zhao, Z G; Zhemchugov, A; Zheng, B; Zheng, J P; Zheng, Y H; Zheng, Z P; Zhong, B; Zhong, J; Zhou, L; Zhou, X K; Zhou, X R; Zhu, C; Zhu, K; Zhu, K J; Zhu, S H; Zhu, X L; Zhu, X W; Zhu, Y M; Zhu, Y S; Zhu, Z A; Zhuang, J; Zou, B S; Zou, J H; Zuo, J X

    2012-10-26

    The two-photon transition ψ(3686)→γγJ/ψ is studied in a sample of 1.06×10(8) ψ(3686) decays collected by the BESIII detector. The branching fraction is measured to be (3.1±0.6(stat)(-1.0)(+0.8)(syst))×10(-4) using J/ψ→e(+)e(-) and J/ψ→μ(+)μ(-) decays, and its upper limit is estimated to be 4.5×10(-4) at the 90% confidence level. This work represents the first measurement of a two-photon transition among charmonium states. The orientation of the ψ(3686) decay plane and the J/ψ polarization in this decay are also studied. In addition, the product branching fractions of sequential E1 transitions ψ(3686)→γχ(cJ) and χ(cJ)→γJ/ψ(J=0,1,2) are reported.

  16. Fluorescence spectroscopy of biological tissue: single- and two-photon excitation

    Science.gov (United States)

    Wu, Yicong; Xi, Peng; Ge, Weikun; Yuen, Powing; Qu, Jianan Y.

    2004-06-01

    Endogenous fluorophores, such as NAD(P)H/FAD and collagen/elastin, have been regarded as in vivo quantitative fluorescence biomarkers for precancerous changes of epithelial tissue. However, the fluorescence signal measured by conventional spectroscopy is a mixture of autofluorescence from the epithelium and deep structures. The dominant fluorescence of collagen/elastin from connective tissue in deep layers creates serious challenge for extracting the epithelial fluorescence of NAD(P)H/FAD that is weak, but important for the characterization of tissue pathology. In this work, we instrumented a confocal fluorescence spectroscopy system and a two-photon excited fluorescence spectroscopy system to measure the depth-resolved single- and two-photon fluorescence spectra from the rabbit esophageal tissues. The excitation wavelengths were 349 nm and 735 nm, respectively. Both systems provided good optical sectioning. The information obtained from depth-resolved fluorescence was generally consistent with the histology of the examined tissue sample. The NAD(P)H signals from epithelial layers were clearly separated from the collagen signal from deep layers. In addition, strong second harmonic generations given by collagen fibers were observed. This work demonstrates that depth-resolved fluorescence spectroscopy may produce more accurate information on the diagnosis of tissue pathology.

  17. Charm and bottom production in two photon collisions at LEP with the L3 detector

    CERN Document Server

    Saremi, S

    2001-01-01

    Inclusive c and b quark production in gamma gamma collisions are studied with the L3 detector at the LEP collider. Data were collected at centre of mass energies from 189 GeV to 202 GeV for a total integrated luminosity of 410 pb/sup -1/. Hadronic final states containing c and b quarks are identified by detecting electrons or muons from their semileptonic decays. The cross section sigma ( gamma gamma to ccX) is measured as a function of the two photon centre of mass energy in the interval 5 GeVtwo photon collisions is measured using the data collected at the centre of mass energy square root s=189 GeV with an...

  18. Two-photon double ionization of neon using an intense attosecond pulse train

    CERN Document Server

    Manschwetus, B; Campi, F; Maclot, S; Coudert-Alteirac, H; Lahl, J; Wikmark, H; Rudawski, P; Heyl, C M; Farkas, B; Mohamed, T; L'Huillier, A; Johnsson, P

    2016-01-01

    We present the first demonstration of two-photon double ionization of neon using an intense extreme ultraviolet (XUV) attosecond pulse train (APT) in a photon energy regime where both direct and sequential mechanisms are allowed. For an APT generated through high-order harmonic generation (HHG) in argon we achieve a total pulse energy close to 1 $\\mu$J, a central energy of 35 eV and a total bandwidth of $\\sim30$ eV. The APT is focused by broadband optics in a neon gas target to an intensity of $3\\cdot10^{12} $W$\\cdot$cm$^{-2}$. By tuning the photon energy across the threshold for the sequential process the double ionization signal can be turned on and off, indicating that the two-photon double ionization predominantly occurs through a sequential process. The demonstrated performance opens up possibilities for future XUV-XUV pump-probe experiments with attosecond temporal resolution in a photon energy range where it is possible to unravel the dynamics behind direct vs. sequential double ionization and the asso...

  19. 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.

  20. Hard Two-Photon Contribution to Elastic Lepton-Proton Scattering Determined by the OLYMPUS Experiment

    Science.gov (United States)

    Henderson, B. S.; Ice, L. D.; Khaneft, D.; O'Connor, C.; Russell, R.; Schmidt, A.; Bernauer, J. C.; Kohl, M.; Akopov, N.; Alarcon, R.; Ates, O.; Avetisyan, A.; Beck, R.; Belostotski, S.; Bessuille, J.; Brinker, F.; Calarco, J. R.; Carassiti, V.; Cisbani, E.; Ciullo, G.; Contalbrigo, M.; de Leo, R.; Diefenbach, J.; Donnelly, T. W.; Dow, K.; Elbakian, G.; Eversheim, P. D.; Frullani, S.; Funke, Ch.; Gavrilov, G.; Gläser, B.; Görrissen, N.; Hasell, D. K.; Hauschildt, J.; Hoffmeister, Ph.; Holler, Y.; Ihloff, E.; Izotov, A.; Kaiser, R.; Karyan, G.; Kelsey, J.; Kiselev, A.; Klassen, P.; Krivshich, A.; Lehmann, I.; Lenisa, P.; Lenz, D.; Lumsden, S.; Ma, Y.; Maas, F.; Marukyan, H.; Miklukho, O.; Milner, R. G.; Movsisyan, A.; Murray, M.; Naryshkin, Y.; Perez Benito, R.; Perrino, R.; Redwine, R. P.; Rodríguez Piñeiro, D.; Rosner, G.; Schneekloth, U.; Seitz, B.; Statera, M.; Thiel, A.; Vardanyan, H.; Veretennikov, D.; Vidal, C.; Winnebeck, A.; Yeganov, V.; Olympus Collaboration

    2017-03-01

    The OLYMPUS Collaboration reports on a precision measurement of the positron-proton to electron-proton elastic cross section ratio, R2 γ , a direct measure of the contribution of hard two-photon exchange to the elastic cross section. In the OLYMPUS measurement, 2.01 GeV electron and positron beams were directed through a hydrogen gas target internal to the DORIS storage ring at DESY. A toroidal magnetic spectrometer instrumented with drift chambers and time-of-flight scintillators detected elastically scattered leptons in coincidence with recoiling protons over a scattering angle range of ≈20 ° to 80°. The relative luminosity between the two beam species was monitored using tracking telescopes of interleaved gas electron multiplier and multiwire proportional chamber detectors at 12°, as well as symmetric Møller or Bhabha calorimeters at 1.29°. A total integrated luminosity of 4.5 fb-1 was collected. In the extraction of R2 γ, radiative effects were taken into account using a Monte Carlo generator to simulate the convolutions of internal bremsstrahlung with experiment-specific conditions such as detector acceptance and reconstruction efficiency. The resulting values of R2 γ, presented here for a wide range of virtual photon polarization 0.456 <ɛ <0.978 , are smaller than some hadronic two-photon exchange calculations predict, but are in reasonable agreement with a subtracted dispersion model and a phenomenological fit to the form factor data.

  1. Hard two-photon contribution to elastic lepton-proton scattering determined by the OLYMPUS experiment

    Energy Technology Data Exchange (ETDEWEB)

    Henderson, B.S. [Massachusetts Institute of Technology, Cambridge, MA (United States); Ice, L.D. [Arizona State Univ., Tempe, AZ (United States); Khaneft, D. [Mainz Univ. (Germany); Collaboration: OLYMPUS Collaboration; and others

    2016-12-15

    The OLYMPUS collaboration reports on a precision measurement of the positron-proton to electron-proton elastic cross section ratio, R{sub 2γ}, a direct measure of the contribution of hard two- photon exchange to the elastic cross section. In the OLYMPUS measurement, 2.01 GeV electron and positron beams were directed through a hydrogen gas target internal to the DORIS storage ring at DESY. A toroidal magnetic spectrometer instrumented with drift chambers and time-of-flight scintillators detected elastically scattered leptons in coincidence with recoiling protons over a scattering angle range of ∼20 to 80 . The relative luminosity between the two beam species was monitored using tracking telescopes of interleaved GEM and MWPC detectors at 12 , as well as symmetric Moeller/Bhabha calorimeters at 1.29 . A total integrated luminosity of 4.5 fb{sup -1} was collected. In the extraction of R{sub 2γ}, radiative effects were taken into account using a Monte Carlo generator to simulate the convolutions of internal bremsstrahlung with experiment-specific conditions such as detector acceptance and reconstruction efficiency. The resulting values of R{sub 2γ}, presented here for a wide range of virtual photon polarization 0.456<ε<0.978, are smaller than some hadronic two-photon exchange calculations predict, but are in reasonable agreement with a subtracted dispersion model and a phenomenological fit to the form factor data.

  2. Resonant two-photon absorption and electromagnetically induced transparency in open ladder-type atomic system.

    Science.gov (United States)

    Moon, Han Seb; Noh, Heung-Ryoul

    2013-03-25

    We have experimentally and theoretically studied resonant two-photon absorption (TPA) and electromagnetically induced transparency (EIT) in the open ladder-type atomic system of the 5S(1/2) (F = 1)-5P(3/2) (F' = 0, 1, 2)-5D(5/2) (F″ = 1, 2, 3) transitions in (87)Rb atoms. As the coupling laser intensity was increased, the resonant TPA was transformed to EIT for the 5S(1/2) (F = 1)-5P(3/2) (F' = 2)-5D(5/2) (F″ = 3) transition. The transformation of resonant TPA into EIT was numerically calculated for various coupling laser intensities, considering all the degenerate magnetic sublevels of the 5S(1/2)-5P(3/2)-5D(5/2) transition. From the numerical results, the crossover from TPA to EIT could be understood by the decomposition of the spectrum into an EIT component owing to the pure two-photon coherence and a TPA component caused by the mixed term.

  3. Intracellular calmodulin availability accessed with two-photon cross-correlation

    Science.gov (United States)

    Kim, Sally A.; Heinze, Katrin G.; Waxham, M. Neal; Schwille, Petra

    2004-01-01

    The availability and interactions of signaling proteins are tightly regulated in time and space to produce specific and localized effects. For calmodulin (CaM), a key transducer of intracellular Ca2+ signaling, binding to its variety of targets initiates signaling cascades and regulates its subcellular localization, thereby making it unavailable for subsequent binding interactions. Among CaM's numerous targets, Ca2+/CaM-dependent protein kinase II is one of the most striking due to its unique ability to increase its affinity for CaM by autophosphorylation and to translocate when bound to Ca2+/CaM. Two-photon fluorescence correlation spectroscopy and cross-correlation spectroscopy were utilized to compare mobility and molecular interactions between CaM and Ca2+/CaM-dependent protein kinase II in solution and in living cells. These techniques revealed that CaM availability in cells could be altered by a change in intracellular conditions. Two-photon fluorescence cross-correlation spectroscopy exemplifies a generally applicable approach for studying protein–protein interactions in living cells that allows access to the behavior of signaling molecules within their native environment to probe for heterogeneities in signaling pathways in different cellular compartments. PMID:14695888

  4. 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.

  5. Double-Tag Events in Two-Photon Collisions at LEP

    CERN Document Server

    Achard, P.; Aguilar-Benitez, M.; Alcaraz, J.; Alemanni, G.; Allaby, J.; Aloisio, A.; Alviggi, M.G.; Anderhub, H.; Andreev, Valery 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.; Barillere, R.; Bartalini, P.; Basile, M.; Batalova, N.; Battiston, R.; Bay, A.; Becattini, F.; Becker, U.; Behner, F.; Bellucci, L.; Berbeco, R.; Berdugo, J.; Berges, P.; Bertucci, B.; Betev, B.L.; Biasini, M.; Biglietti, M.; Biland, A.; Blaising, J.J.; Blyth, S.C.; Bobbink, G.J.; Bohm, A.; Boldizsar, L.; Borgia, B.; Bottai, S.; Bourilkov, D.; Bourquin, M.; Braccini, S.; Branson, J.G.; Brochu, F.; Buijs, A.; Burger, J.D.; Burger, W.J.; Cai, X.D.; Capell, M.; Cara Romeo, G.; Carlino, G.; Cartacci, A.; Casaus, J.; Cavallari, F.; Cavallo, N.; Cecchi, C.; Cerrada, M.; Chamizo, M.; Chang, Y.H.; Chemarin, M.; Chen, A.; Chen, G.; Chen, G.M.; Chen, H.F.; Chen, H.S.; Chiefari, G.; Cifarelli, L.; Cindolo, F.; Clare, I.; Clare, R.; Coignet, G.; Colino, N.; Costantini, S.; de la Cruz, B.; Cucciarelli, S.; van Dalen, J.A.; de Asmundis, R.; Deglon, P.; Debreczeni, J.; Degre, A.; Deiters, K.; della Volpe, D.; Delmeire, E.; Denes, P.; DeNotaristefani, F.; De Salvo, A.; Diemoz, M.; Dierckxsens, M.; van Dierendonck, D.; Dionisi, C.; Dittmar, M.; Doria, A.; Dova, M.T.; Duchesneau, D.; Duinker, P.; Echenard, B.; Eline, A.; El Mamouni, H.; Engler, A.; Eppling, F.J.; Ewers, A.; Extermann, P.; Falagan, M.A.; Falciano, S.; Favara, A.; Fay, J.; Fedin, O.; Felcini, M.; Ferguson, T.; Fesefeldt, H.; Fiandrini, E.; Field, J.H.; Filthaut, F.; Fisher, P.H.; Fisher, W.; Fisk, I.; Forconi, G.; Freudenreich, K.; Furetta, C.; Galaktionov, Iouri; Ganguli, S.N.; Garcia-Abia, Pablo; Gataullin, M.; Gentile, S.; Giagu, S.; Gong, Z.F.; Grenier, Gerald Jean; Grimm, O.; Gruenewald, M.W.; Guida, M.; van Gulik, R.; Gupta, V.K.; Gurtu, A.; Gutay, L.J.; Haas, D.; Hatzifotiadou, D.; Hebbeker, T.; Herve, Alain; Hirschfelder, J.; Hofer, H.; Hohlmann, M.; Holzner, G.; Hou, S.R.; Hu, Y.; Jin, B.N.; Jones, Lawrence W.; de Jong, P.; Josa-Mutuberria, I.; Kafer, D.; Kaur, M.; Kienzle-Focacci, M.N.; Kim, J.K.; Kirkby, Jasper; Kittel, W.; Klimentov, A.; Konig, A.C.; Kopal, M.; Koutsenko, V.; Kraber, M.; Kraemer, R.W.; Krenz, W.; Kruger, A.; Kunin, A.; Ladron 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, P.; Li, C.; Likhoded, S.; Lin, C.H.; Lin, W.T.; Linde, F.L.; Lista, L.; Liu, Z.A.; Lohmann, W.; Longo, E.; Lu, Y.S.; Lubelsmeyer, K.; Luci, C.; Luminari, L.; Lustermann, W.; Ma, W.G.; Malgeri, L.; Malinin, A.; Mana, C.; Mangeol, D.; 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.; Kluge, Hannelies; Ofierzynski, R.; Organtini, G.; Palomares, C.; Pandoulas, D.; Paolucci, P.; Paramatti, R.; Passaleva, G.; Patricelli, S.; Paul, Thomas Cantzon; Pauluzzi, M.; Paus, C.; Pauss, F.; Pedace, M.; Pensotti, S.; Perret-Gallix, D.; Petersen, B.; Piccolo, D.; Pierella, F.; Pioppi, M.; Piroue, P.A.; Pistolesi, E.; Plyaskin, V.; Pohl, M.; Pojidaev, V.; Pothier, J.; Prokofev, D.O.; Prokofiev, D.; Quartieri, J.; Rahal-Callot, G.; Rahaman, M.A.; Raics, P.; Raja, N.; Ramelli, R.; Rancoita, P.G.; Ranieri, R.; Raspereza, A.; Razis, P.; Ren, D.; Rescigno, M.; Reucroft, S.; Riemann, S.; Riles, Keith; Roe, B.P.; Romero, L.; Rosca, A.; Rosier-Lees, S.; Roth, Stefan; Rosenbleck, C.; Roux, B.; Rubio, J.A.; Ruggiero, G.; Rykaczewski, H.; Sakharov, A.; Saremi, S.; Sarkar, S.; Salicio, J.; Sanchez, E.; Sanders, M.P.; Schafer, C.; Schegelsky, V.; Schmidt-Kaerst, S.; Schmitz, D.; Schopper, H.; Schotanus, D.J.; Schwering, G.; Sciacca, C.; Servoli, L.; Shevchenko, S.; Shivarov, N.; Shoutko, V.; Shumilov, E.; Shvorob, A.; Siedenburg, T.; Son, D.; Spillantini, P.; Steuer, M.; Stickland, D.P.; Stoyanov, B.; Straessner, A.; Sudhakar, K.; Sultanov, G.; Sun, L.Z.; Sushkov, S.; Suter, H.; Swain, J.D.; Szillasi, Z.; Tang, X.W.; Tarjan, P.; Tauscher, L.; Taylor, L.; Tellili, B.; Teyssier, D.; Timmermans, Charles; Ting, Samuel C.C.; Ting, S.M.; Tonwar, S.C.; Toth, J.; Tully, C.; Tung, K.L.; Ulbricht, J.; Valente, E.; Van de Walle, R.T.; Veszpremi, V.; Vesztergombi, G.; Vetlitsky, I.; Vicinanza, D.; Viertel, G.; Villa, S.; Vivargent, M.; Vlachos, S.; Vodopianov, 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.; Zoller, M.

    2002-01-01

    Double-tag events in two-photon collisions are studied using the L3 detector at LEP centre-of-mass energies from root(s)=189 GeV to 209 GeV. The cross sections of the e+e- -> e+e- hadrons and gamma*gamma* -> hadrons processes are measured as a function of the photon virtualities, Q2_1 and Q2_2, of the two-photon mass, W_gammagamma, and of the variable Y=ln(W2_gammagamma/(Q_1Q_2)), for an average photon virtuality = 16 GeV2. The results are in agreement with next-to-leading order calculations for the process gamma*gamma* -> q qbar in the interval 2 <= Y <= 5. An excess is observed in the interval 5 < Y <= 7, corresponding to W_gammagamma greater than 40 GeV . This may be interpreted as a sign of resolved photon QCD processes or the onset of BFKL phenomena.

  6. Size Dependence of Two-Photon Absorption in Semiconductor Quantum Dots

    Energy Technology Data Exchange (ETDEWEB)

    Dakovski, Georgi L. [Case Western Reserve Univ., Cleveland, OH (United States); Shan, Jie [Case Western Reserve Univ., Cleveland, OH (United States)

    2013-01-01

    Quantum confinement plays an important role in the optical properties of semiconductor quantum dots (QDs). In this work, we combine experiment and modeling to systematically investigate the size dependence of the degenerate two-photon absorption (TPA) of below-band-gap radiation in CdSe QDs. The TPA coefficient β at 800 nm of CdSe QDs of varying radii was measured using femtosecond white-light transient absorption spectroscopy by probing the pump-induced bleaching at the first exciton transition energy. β was also calculated using a model based on the multiband effective-mass approximation. Satisfactory agreement between experiment and theory was obtained. Our findings show the evolution of the TPA in the QDs from that of atom-like to bulk-like with increasing the radius R. The TPA coefficient (or the volume normalized TPA cross-section) increases with radius approximately linearly in the strong confinement regime due to the rapid increase of the joint density of states for the two-photon allowed transitions, and saturates for R > 5 nm (the exciton Bohr radius), approaching that of bulk CdSe.

  7. A Nanosystem Capable of Releasing a Photosensitizer Bioprecursor under Two-Photon Irradiation for Photodynamic Therapy.

    Science.gov (United States)

    Wu, Hao; Zeng, Fang; Zhang, Hang; Xu, Jiangsheng; Qiu, Jianrong; Wu, Shuizhu

    2016-02-01

    The applications of photodynamic therapy (PDT) are usually limited by photosensitizers' side effects and singlet oxygen's short half-life. Herein, a mitochondria-targeted nanosystem is demonstrated to enhance the PDT efficacy by releasing a bio-precursor of photosensitizer under two-photon irradiation. A phototriggerable coumarin derivative is first synthesized by linking 5-aminolevulinic acid (5-ALA, the bio-precursor) to coumarin; and the nanosystem (CD-ALA-TPP) is then fabricated by covalently incorporating this coumarin derivative and a mitochondria-targeting compound triphenylphosphonium (TPP) onto carbon dots (CDs). Upon cellular internalization, the nanosystem preferentially accumulates in mitochondria; and under one- or two-photon irradiation, it releases 5-ALA molecules that are then metabolized into protoporphyrin IX in mitochondria through a series of biosynthesis processes. The subsequent red light irradiation induces this endogenously synthesized photosensitizer to generate singlet oxygen, thereby causing oxidant damage to mitochondria and then the apoptosis of the cells. Analysis via 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) assays indicate that the novel PDT system exhibits enhanced cytotoxicity toward cancer cells. This study may offer a new strategy for designing PDT systems with high efficacy and low side effects.

  8. Concise synthesis and two-photon-excited deep-blue emission of 1,8-diazapyrenes.

    Science.gov (United States)

    He, Tingchao; Too, Pei Chui; Chen, Rui; Chiba, Shunsuke; Sun, Handong

    2012-09-01

    Efficient violet-blue-emitting molecules are especially useful for applications in full-color displays, solid-state lighting, as well as in two-photon absorption (TPA) excited frequency-upconverted violet-blue lasing. However, the reported violet-blue-emitting molecules generally possess small TPA cross sections. In this work, new 1,8-diazapyrenes derivatives 3 with blue two-photon-excited fluorescence emission were concisely synthesized by the coupling reaction of readily available 1,4-naphthoquinone O,O-diacetyl dioxime (1) with internal alkynes 2 under the [{RhCl(2)Cp*}(2)]-Cu(OAc)(2) (Cp*=pentamethylcyclopentadienyl ligand) bimetallic catalytic system. Elongation of the π-conjugated length of 1,8-diazapyrenes 3 led to the increase of TPA cross sections without the expense of a redshift of the emission wavelength, probably due to the rigid planar structure of chromophores. It is especially noteworthy that 2,3,6,7-tetra(4-bromophenyl)-1,8-diazapyrene (3c) has a larger TPA cross section than those of other molecules reported so far. These experimental results are explained in terms of the effects of extension of the π-conjugated system, intramolecular charge transfer, and reduced detuning energy. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Charge Transport in Two-Photon Semiconducting Structures for Solar Fuels.

    Science.gov (United States)

    Liu, Guohua; Du, Kang; Haussener, Sophia; Wang, Kaiying

    2016-10-20

    Semiconducting heterostructures are emerging as promising light absorbers and offer effective electron-hole separation to drive solar chemistry. This technology relies on semiconductor composites or photoelectrodes that work in the presence of a redox mediator and that create cascade junctions to promote surface catalytic reactions. Rational tuning of their structures and compositions is crucial to fully exploit their functionality. In this review, we describe the possibilities of applying the two-photon concept to the field of solar fuels. A wide range of strategies including the indirect combination of two semiconductors by a redox couple, direct coupling of two semiconductors, multicomponent structures with a conductive mediator, related photoelectrodes, as well as two-photon cells are discussed for light energy harvesting and charge transport. Examples of charge extraction models from the literature are summarized to understand the mechanism of interfacial carrier dynamics and to rationalize experimental observations. We focus on a working principle of the constituent components and linking the photosynthetic activity with the proposed models. This work gives a new perspective on artificial photosynthesis by taking simultaneous advantages of photon absorption and charge transfer, outlining an encouraging roadmap towards solar fuels. © 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Efficient Entanglement Concentration of Nonlocal Two-Photon Polarization-Time-Bin Hyperentangled States

    Science.gov (United States)

    Wang, Zi-Hang; Yu, Wen-Xuan; Wu, Xiao-Yuan; Gao, Cheng-Yan; Alzahrani, Faris; Hobiny, Aatef; Deng, Fu-Guo

    2017-11-01

    We present two different hyperentanglement concentration protocols (hyper-ECPs) for two-photon systems in nonlocal polarization-time-bin hyperentangled states with known parameters, including Bell-like and cluster-like states, resorting to the parameter splitting method. They require only one of two parties in quantum communication to operate her photon in the process of entanglement concentration, not two, and they have the maximal success probability. They work with linear optical elements and have good feasibility in experiment, especially in the case that there are a big number of quantum data exchanged as the parties can obtain the information about the parameters of the nonlocal hyperentangled states by sampling a subset of nonlocal hyperentangled two-photon systems and measuring them. As the quantum state of photons in the time-bin degree of freedom suffers from less noise in an optical-fiber channel, these hyper-ECPs may have good applications in practical long-distance quantum communication in the future.

  11. Surface plasmon-enhanced and quenched two-photon excited fluorescence

    Science.gov (United States)

    Lin, C.-Y.; Lien, C.-H.; Chiu, K.-C.; Chang, C.-Y.; Chang, S.-H.; Guo, T.-F.; Chen, S.-J.

    2010-08-01

    This study investigated theoretically and experimentally that two-photon excited fluorescence is enhanced and quenched via surface plasmons (SPs) excited by total internal reflection with a silver film. The fluorescence intensity is fundamentally affected by the local electromagnetic field enhancement and the quantum yield change according to the surrounding structure and materials. By utilizing the Fresnel equation and classical dipole radiation modeling, local electric field enhancement, fluorescence quantum yield, and fluorescence emission coupling yield via SPs were theoretically analyzed at different dielectric spacer thicknesses between the fluorescence dye and the metal film. The fluorescence lifetime was also decreased substantially via the quenching effect. A two-photon excited total internal reflection fluorescence (TIRF) microscopy with a time-correlated single photon counting device has been developed to measure the fluorescence lifetimes, photostabilities, and enhancements. The experimental results demonstrate that the fluorescence lifetimes and the trend of the enhancements are consistent with the theoretical analysis. The maximum fluorescence enhancement factor in the surface plasmon-total internal reflection fluorescence (SP-TIRF) configuration can be increased up to 30 fold with a suitable thickness SiO2 spacer. Also, to compromise for the fluorescence enhancement and the fluorophore photostability, we find that the SP-TIRF configuration with a 10 nm SiO2 spacer can provide an enhanced and less photobleached fluorescent signal via the assistance of enhanced local electromagnetic field and quenched fluorescence lifetime, respectively.

  12. Two-photon absorption and upconversion luminescence of colloidal CsPbX3 quantum dots

    Science.gov (United States)

    Han, Qiuju; Wu, Wenzhi; Liu, Weilong; Yang, Qingxin; Yang, Yanqiang

    2018-01-01

    The nonlinear optical and the upconversion luminescence (UCL) properties of CsPbX3 (X = Br or its binary mixtures with Cl, I) quantum dots (QDs) are investigated by femtosecond open-aperture (OA) Z-scan and time-resolved luminescence techniques in nonresonant spectral region. The OA Z-scan results show that CsPbX3 QDs have strong reverse saturable absorption (RSA), which is ascribed to two-photon absorption. Partially changing halide composition from Cl to Br, to I, two-photon absorption cross sections become larger at the same laser excitation intensity. The composition-tunable nonlinear absorption should be attributed to the gradual decrease of the lowest direct band gaps with the halide substitute. Moreover, the strong UCL can be observed under near infrared femtosecond laser excitation. Halide composition-tunable UCL dynamics of CsPbX3 QDs is analyzed by use of two-exponential fitting with deconvolution. When CsPbX3 QDs have similar sizes (10-13 nm), with partially changing halide composition from Cl to Br, to I, the average UCL lifetime becomes longer due to the variation of Kane energy. Our findings suggest all-inorganic perovskite QDs can be used as excellent gain medium for high-performance frequency-upconversion lasers and provide reference to engineer such QDs toward practical optoelectronic applications.

  13. Two-Photon Optical Storage in Photorefractive Polymers in the Near-Infrared Spectral Range

    Science.gov (United States)

    Day, Daniel; Gu, Min; Smallridge, Andrew

    We report the use of a polymer-based photorefractive material for three-dimensional bit optical data storage using near-infrared illumination. The research was conducted using photorefractive materials that were fabricated in two polymer matrices: poly(N-vinylcarbazole) (PVK) and poly(Methyl Methacrylate) (PMMA). The recording samples also consisted of the following compounds in various proportions: 2,5-dimethyl-4-(p-nitrophenylazo)anisole (DMNPAA), 2,4,7-trinitro-9-fluorenone (TNF) and N-ethylcarbazole (ECZ). Two-photon excitation was used as the recording mechanism to achieve rewritable bit data storage in a photorefractive polymer. As a result of two-photon excitation, the quadratic dependence of the excitation on the incident intensity produces an excitation volume that is confined to the focal region in both the transverse and axial directions. The use of ultrashort pulsed lasers, while effective, is not a practical solution for an optical data storage system. This research demonstrates the ability to produce three-dimensional rewritable bit data storage using continuous-wave illumination. Using this technology it has been possible to achieve a density of 88 Gbits/cm3, which in the future could be increased to 3.5 Tbits/cm3.

  14. An ESIPT-based two-photon fluorescent probe detection of hydrogen peroxide in live cells and tissues.

    Science.gov (United States)

    Zhou, Liyi; Peng, Yongbo; Wang, Qianqian; Lin, Qinlu

    2017-02-01

    A variety of diseases associated with human aging, which have a strong oxidative stress, but connecting age-related diseases and oxidative stress of the basic molecular mechanisms still insufficiently understood. Oxidative stress origins from the unregulated production of reactive oxygen species (ROS), and oxidative damaging to tissues and organs from subsequent oxidation-reduction chemistry by cellular mismanagement. In particular, H2O2 is a major by-product of ROS in live organisms and a common marker for oxidative stress, and its dynamic equilibrium can have various physiological and pathological consequences. H2O2 is a small molecule, but it is an essential oxygen metabolite in living systems and acts as an important compound in cellular signal transduction by reversible oxidation of proteins. To quantitatively detect of H2O2 in biosystems, herein, we adopted a 2-(2'-hydroxyphenyl)-4(3H)-quinazolinone (HPQ), a small organic fluorophore known for its luminescence mechanism through excited-state intramolecular proton transfer (ESIPT). HPQ was employed as a precursor to develop a turn-on probe (HPQ-H) for bioimaging applications. After cleavaging the boronic ester moiety by H2O2, HPQ-H releases a HPQ fluorophore which shows a 45-fold fluorescence intensity enhancement with high sensitivity and selectivity over other reactive oxygen species (ROS), and a high resolution imaging and large tissue-imaging depth (70-170μm) in living cells and tissues images under two-photon excitation (720nm). Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Strong-field QED processes in short laser pulses. One- and two-photon Compton scattering

    Energy Technology Data Exchange (ETDEWEB)

    Seipt, Daniel

    2012-12-20

    The purpose of this thesis is to advance the understanding of strong-field QED processes in short laser pulses. The processes of non-linear one-photon and two-photon Compton scattering are studied, that is the scattering of photons in the interaction of relativistic electrons with ultra-short high-intensity laser pulses. These investigations are done in view of the present and next generation of ultra-high intensity optical lasers which are supposed to achieve unprecedented intensities of the order of 10{sup 24} W/cm{sup 2} and beyond, with pulse lengths in the order of some femtoseconds. The ultra-high laser intensity requires a non-perturbative description of the interaction of charged particles with the laser field to allow for multi-photon interactions, which is beyond the usual perturbative expansion of QED organized in powers of the fine structure constant. This is achieved in strong-field QED by employing the Furry picture and non-perturbative solutions of the Dirac equation in the presence of a background laser field as initial and final state wave functions, as well as the laser dressed Dirac-Volkov propagator. The primary objective is a realistic description of scattering processes with regard to the finite laser pulse duration beyond the common approximation of infinite plane waves, which is made necessary by the ultra-short pulse length of modern high-intensity lasers. Non-linear finite size effects are identified, which are a result of the interplay between the ultra-high intensity and the ultra-short pulse length. In particular, the frequency spectra and azimuthal photon emission spectra are studied emphasizing the differences between pulsed and infinite laser fields. The proper description of the finite temporal duration of the laser pulse leads to a regularization of unphysical infinities (due to the infinite plane-wave description) of the laser-dressed Dirac-Volkov propagator and in the second-order strong-field process of two-photon Compton

  16. Two-photon polymerization as a structuring technology in production: future or fiction?

    Science.gov (United States)

    Harnisch, Emely Marie; Schmitt, Robert

    2017-02-01

    Two-photon polymerization (TPP) has become an established generative fabrication technique for individual, up to three-dimensional micro- and nanostructures. Due to its high resolution beyond the diffraction limit, its writing speed is limited and in most cases, very special structures are fabricated in small quantities. With regard to the trends of the optical market towards higher efficiencies, miniaturization and higher functionalities, there is a high demand for so called intelligent light management systems, including also individual optical elements. Here, TPP could offer a fabrication technique, enabling higher complexities of structures than conventional cutting and lithographic technologies do. But how can TPP opened up for production? In the following, some approaches to establish TPP as a mastering technique for molding are presented against this background.

  17. f$_1$(1285) Formation in Two-Photon Collisions at LEP

    CERN Document Server

    Achard, P.; Aguilar-Benitez, M.; Alcaraz, J.; Alemanni, G.; Allaby, J.; Aloisio, A.; Alviggi, M.G.; Anderhub, H.; Andreev, Valery 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.; Barillere, R.; Bartalini, P.; Basile, M.; Batalova, N.; Battiston, R.; Bay, A.; Becattini, F.; Becker, U.; Behner, F.; Bellucci, L.; Berbeco, R.; Berdugo, J.; Berges, P.; Bertucci, B.; Betev, B.L.; Biasini, M.; Biglietti, M.; Biland, A.; Blaising, J.J.; Blyth, S.C.; Bobbink, G.J.; Bohm, A.; Boldizsar, L.; Borgia, B.; Bottai, S.; Bourilkov, D.; Bourquin, M.; Braccini, S.; Branson, J.G.; Brochu, F.; Buijs, A.; Burger, J.D.; Burger, W.J.; Cai, X.D.; Capell, M.; Cara Romeo, G.; Carlino, G.; Cartacci, A.; Casaus, J.; Cavallari, F.; Cavallo, N.; Cecchi, C.; Cerrada, M.; Chamizo, M.; Chang, Y.H.; Chemarin, M.; Chen, A.; Chen, G.; Chen, G.M.; Chen, H.F.; Chen, H.S.; Chiefari, G.; Cifarelli, L.; Cindolo, F.; Clare, I.; Clare, R.; Coignet, G.; Colino, N.; Costantini, S.; de la Cruz, B.; Cucciarelli, S.; van Dalen, J.A.; de Asmundis, R.; Deglon, P.; Debreczeni, J.; Degre, A.; Deiters, K.; della Volpe, D.; Delmeire, E.; Denes, P.; DeNotaristefani, F.; De Salvo, A.; Diemoz, M.; Dierckxsens, M.; van Dierendonck, D.; Dionisi, C.; Dittmar, M.; Doria, A.; Dova, M.T.; Duchesneau, D.; Duinker, P.; Echenard, B.; Eline, A.; El Mamouni, H.; Engler, A.; Eppling, F.J.; Ewers, A.; Extermann, P.; Falagan, M.A.; Falciano, S.; Favara, A.; Fay, J.; Fedin, O.; Felcini, M.; Ferguson, T.; Fesefeldt, H.; Fiandrini, E.; Field, J.H.; Filthaut, F.; Fisher, P.H.; Fisher, W.; Fisk, I.; Forconi, G.; Freudenreich, K.; Furetta, C.; Galaktionov, Iouri; Ganguli, S.N.; Garcia-Abia, Pablo; Gataullin, M.; Gentile, S.; Giagu, S.; Gong, Z.F.; Grenier, Gerald Jean; Grimm, O.; Gruenewald, M.W.; Guida, M.; van Gulik, R.; Gupta, V.K.; Gurtu, A.; Gutay, L.J.; Haas, D.; Hatzifotiadou, D.; Hebbeker, T.; Herve, Alain; Hirschfelder, J.; Hofer, H.; Hohlmann, M.; Holzner, G.; Hou, S.R.; Hu, Y.; Jin, B.N.; Jones, Lawrence W.; de Jong, P.; Josa-Mutuberria, I.; Kafer, D.; Kaur, M.; Kienzle-Focacci, M.N.; Kim, J.K.; Kirkby, Jasper; Kittel, W.; Klimentov, A.; Konig, A.C.; Kopal, M.; Koutsenko, V.; Kraber, M.; Kraemer, R.W.; Krenz, W.; Kruger, A.; Kunin, A.; Ladron 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, P.; Li, C.; Likhoded, S.; Lin, C.H.; Lin, W.T.; Linde, F.L.; Lista, L.; Liu, Z.A.; Lohmann, W.; Longo, E.; Lu, Y.S.; Lubelsmeyer, K.; Luci, C.; Luminari, L.; Lustermann, W.; Ma, W.G.; Malgeri, L.; Malinin, A.; Mana, C.; Mangeol, D.; 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.; Kluge, Hannelies; Ofierzynski, R.; Organtini, G.; Palomares, C.; Pandoulas, D.; Paolucci, P.; Paramatti, R.; Passaleva, G.; Patricelli, S.; Paul, Thomas Cantzon; Pauluzzi, M.; Paus, C.; Pauss, F.; Pedace, M.; Pensotti, S.; Perret-Gallix, D.; Petersen, B.; Piccolo, D.; Pierella, F.; Pioppi, M.; Piroue, P.A.; Pistolesi, E.; Plyaskin, V.; Pohl, M.; Pojidaev, V.; Pothier, J.; Prokofev, D.O.; Prokofiev, D.; Quartieri, J.; Rahal-Callot, G.; Rahaman, M.A.; Raics, P.; Raja, N.; Ramelli, R.; Rancoita, P.G.; Ranieri, R.; Raspereza, A.; Razis, P.; Ren, D.; Rescigno, M.; Reucroft, S.; Riemann, S.; Riles, Keith; Roe, B.P.; Romero, L.; Rosca, A.; Rosier-Lees, S.; Roth, Stefan; Rosenbleck, C.; Roux, B.; Rubio, J.A.; Ruggiero, G.; Rykaczewski, H.; Sakharov, A.; Saremi, S.; Sarkar, S.; Salicio, J.; Sanchez, E.; Sanders, M.P.; Schafer, C.; Schegelsky, V.; Schmidt-Kaerst, S.; Schmitz, D.; Schopper, H.; Schotanus, D.J.; Schwering, G.; Sciacca, C.; Servoli, L.; Shevchenko, S.; Shivarov, N.; Shoutko, V.; Shumilov, E.; Shvorob, A.; Siedenburg, T.; Son, D.; Spillantini, P.; Steuer, M.; Stickland, D.P.; Stoyanov, B.; Straessner, A.; Sudhakar, K.; Sultanov, G.; Sun, L.Z.; Sushkov, S.; Suter, H.; Swain, J.D.; Szillasi, Z.; Tang, X.W.; Tarjan, P.; Tauscher, L.; Taylor, L.; Tellili, B.; Teyssier, D.; Timmermans, Charles; Ting, Samuel C.C.; Ting, S.M.; Tonwar, S.C.; Toth, J.; Tully, C.; Tung, K.L.; Ulbricht, J.; Valente, E.; Van de Walle, R.T.; Veszpremi, V.; Vesztergombi, G.; Vetlitsky, I.; Vicinanza, D.; Viertel, G.; Villa, S.; Vivargent, M.; Vlachos, S.; Vodopianov, 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.; Zoller, M.

    2002-01-01

    The $\\eta \\pi^+ \\pi^-$ final state in two-photon collisions is studied with the L3 detector at LEP, at centre-of-mass energies from 183 to 209~GeV with an integrated luminosity of 664.6~pb$^{-1}$. The f$_1$(1285) meson is observed and the $Q^2$ dependence of its production is compared to different form factor models. The $\\gamma\\gamma$-coupling parameter $\\tilde\\Gamma_{\\gamma\\gamma}$ is found to be $3.5 \\pm 0.6\\,(stat.) \\pm 0.5\\,(sys.)$~keV. The branching fraction $\\Gamma\\bigl({\\rm f}_1(1285)\\rightarrow{\\rm a}_0\\pi\\bigr) / \\Gamma\\bigl({\\rm f}_1(1285)\\rightarrow\\eta\\pi\\pi\\bigr)$ is also measured.

  18. Inclusive $D^{*\\pm}$ 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; 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; Carr-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, L; 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; Mazumdari, 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, A; 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 de Walle, R T; 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 Zhao Min; 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*/sup +or-/ production in two-photon collisions is studied with the L3 detector at LEP, using 683 pb/sup -1/ of data collected at centre-of-mass energies from 183 to 209 GeV. Differential cross sections are determined as functions of the transverse momentum and pseudorapidity of the D*/sup +or-/ mesons in the kinematic region 1 GeV

  19. Optical assembly of microsnap-fits fabricated by two-photon polymerization

    Science.gov (United States)

    Köhler, Jannis; Kutlu, Yunus; Zyla, Gordon; Ksouri, Sarah I.; Esen, Cemal; Gurevich, Evgeny L.; Ostendorf, Andreas

    2017-10-01

    To respond to current demands of nano- and microtechnologies, e.g., miniaturization and integration, different bottom-up strategies have been developed. These strategies are based on picking, placing, and assembly of multiple components to produce microsystems with desired features. This paper covers the fabrication of arbitrary-shaped microcomponents by two-photon polymerization and the trapping, moving, and aligning of these structures by the use of a holographic optical tweezer. The main focus is on the assembly technique based on a cantilever microsnap-fit. More precisely, mechanical properties are characterized by optical forces and a suitable geometry of the snap-fit is designed. As a result of these investigations, a fast and simple assembly technique is developed. Furthermore, disassembly is provided by an optimized design. These findings suggest that the microsnap-fit is suitable for the assembly of miniaturized systems and could broaden the application opportunities of bottom-up strategies.

  20. Charm production in two-photon collisions measured by the ALEPH detector at LEP II energies

    CERN Document Server

    Sieler, U

    2001-01-01

    Charm production in two-photon collisions has been measured at LEP for center of mass energies of square root S/sub e//sup +//sub e//sup -/=183-189 GeV using the ALEPH detector. Charm events have been detected via muons from semi leptonic decays and via inclusive D/sup */ production. Preliminary results are presented. The total cross section sigma (e/sup +/e/sup -/ to e/sup +/e/sup -/cc) has been measured. The relative fractions of the contributing processes have been determined for the acceptance range of D/sup */ mesons. Differential D/sup */ cross sections in the transverse momentum and the pseudorapidity of the D/sup */'s have been measured and compared to NLO QCD calculations. (14 refs).

  1. Leptonic two-photon events measured with L3 at the Large Electron Positron Collider (LEP)

    CERN Document Server

    Dehmelt, Klaus

    2006-01-01

    Virtual photons can fluctuate into diverse final states and among other processes, such fluctuations can yield muon-pairs. The fluctuation is a statistical process and can be described in terms of structure functions for photons. Apart from supplying another test of QED, purely leptonic processes provide a calibration fro the hadronic processes. We report on a measurement of dimuon events with single-tagged two-photon events from the LEP experiment L3, at c.m.s. energies between 189 GeV and 206 GeV, with an integrated luminosity of $\\mathcal{L}$ = 600 pb$^{-1}$. An overview of the fundamental physical processes, the detector and the anaytical tools, used for measuring dimuon events, and a discussion of the structure of dimuon events will be presented.

  2. Two-photon quantum interference in integrated multi-mode interference devices.

    Science.gov (United States)

    Poulios, Konstantinos; Fry, Daniel; Politi, Alberto; Ismail, Nur; Wörhoff, Kerstin; O'Brien, Jeremy L; Thompson, Mark G

    2013-10-07

    Multi-mode interference (MMI) devices fabricated in silicon oxynitride (SiON) with a refractive index contrast of 2.4% provide a highly compact and stable platform for multi-photon non-classical interference. MMI devices can introduce which-path information for photons propagating in the multi-mode section which can result in degradation of this non-classical interference. We theoretically derive the visibility of quantum interference of two photons injected in a MMI device and predict near unity visibility for compact SiON devices. We complement the theoretical results by experimentally demonstrating visibilities of up to 97.7% in 2×2 MMI devices without the requirement of narrow-band photons.

  3. ACTIVE MEDIA: Two-photon Raman processes in spectral ultrabroadening of intense ultrashort light pulses

    Science.gov (United States)

    Belenov, É. M.; Prokopovich, I. P.

    1993-06-01

    The spectral evolution of an intense electromagnetic pulse in the course of a coherent two-photon Raman interaction with a medium is analyzed under conditions such that the approximation of slow envelopes is not valid for the field or polarization of the medium. There exist threshold power levels at which a pronounced self-broadening of the field spectrum occurs if the field spectrum includes the frequencies Ω, where \\hslashΩ=E2 - E1 is the energy difference between the levels involved in the stimulated Raman scattering. The results found here agree with experimental data on the generation of a supercontinuum in various media. In particular, there is agreement in terms of the spectral self-transformation of optical solitons in optical fibers.

  4. Two-photon excited fluorescence of the lens for the diagnosis of presbyopia

    Science.gov (United States)

    Steiner, R.; Kessler, M.; Fugger, O.; Dolp, F.; Russ, D.

    2009-09-01

    Presbyopia is a wide spread phenomenon in elder people and is caused by the hardening of the lens in human eyes. Research is performed to make such lenses again more flexible by application of geometrically optimised cuts through the lens with a femtosecond-laser. Different protein agglomerations are responsible for the flexibility reduction of the lens. Two-photon excited fluorescence of the lens can be used as a diagnostic tool to localise such protein accumulations. In in-vitro experiments with human cataract lenses and also lenses of the Philly-mouse it could be demonstrated that with age the fluorescence increases as presbyopia proceeds. The distribution of the fluorescing compounds are not homogeneous but rather cloudy. Discrimination of the compounds by fluorescence lifetime measurements in relation of the depth in the lens is possible.

  5. Blu-ray disk lens as the objective of a miniaturized two-photon fluorescence microscope.

    Science.gov (United States)

    Chung, Hsiang-Yu; Kuo, Wei-Cheng; Cheng, Yu-Hsiang; Yu, Che-Hang; Chia, Shih-Hsuan; Lin, Cheng-Yung; Chen, Jie-Shin; Tsai, Huai-Jen; Fedotov, Andrey B; Ivanov, Anatoly A; Zheltikov, Aleksei M; Sun, Chi-Kuang

    2013-12-16

    In this paper, we examine the performance of a Blu-ray disk (BD) aspheric lens as the objective of a miniaturized scanning nonlinear optical microscope. By combining a single 2D micro-electro mechanical system (MEMS) mirror as the scanner and with different tube lens pairs, the field of view (FOV) of the studied microscope varies from 59 μm × 93 μm up to 178 μm × 280 μm, while the corresponding lateral resolution varies from 0.6 μm to 2 μm for two-photon fluorescence (2PF) signals. With a 34/s video frame rate, in vivo dynamic observation of zebrafish heartbeat through 2PF of the excited green fluorescence protein (GFP) is demonstrated.

  6. Search for a two-photon exchange contribution to inclusive deep-inelastic scattering

    Energy Technology Data Exchange (ETDEWEB)

    Airapetian, A. [Giessen Univ. (Germany). Physikalisches Institut; Michigan Univ., Ann Arbor (United States). Randall Lab. of Physics; Akopov, N. [Yerevan Physics Institute (Argentina); Akopov, Z. [DESY Hamburg (DE)] (and others)

    2009-07-15

    The transverse-target single-spin asymmetry for inclusive deep-inelastic scattering with effectively unpolarized electron and positron beams off a transversely polarized hydrogen target was measured, with the goal of searching for a two-photon exchange signal in the kinematic range 0.0071 GeV{sup 2} and Q{sup 2}<1 GeV{sup 2}, and for both electron and positron beams, the asymmetries are found to be consistent with zero within statistical and systematic uncertainties, which are of order 10{sup -3} for the asymmetries integrated over x{sub B}. (orig.)

  7. The $\\eta_c$(2980) formation in two-photon collisions at LEP energies

    CERN Document Server

    Abdallah, J; Adam, W; Adzic, P; Albrecht, T; Alderweireld, T; Alemany-Fernandez, R; Allmendinger, T; Allport, P P; Amaldi, Ugo; Amapane, N; Amato, S; Anashkin, E; Andreazza, A; Andringa, S; Anjos, N; Antilogus, P; Apel, W D; Arnoud, Y; Ask, S; Åsman, B; Augustin, J E; Augustinus, A; Baillon, Paul; Ballestrero, A; Bambade, P; Barbier, R; Bardin, Dimitri Yuri; Barker, G; Baroncelli, A; Battaglia, Marco; Baubillier, M; Becks, K H; Begalli, M; Behrmann, A; Ben-Haim, E; Benekos, N C; Benvenuti, Alberto C; Bérat, C; Berggren, M; Berntzon, L; Bertrand, D; Besançon, M; Besson, N; Bloch, D; Blom, M; Bluj, M; Bonesini, M; Boonekamp, M; Booth, P S L; Borisov, G; Botner, O; Bouquet, B; Bowcock, T J V; Boyko, I; Bracko, M; Brenner, R; Brodet, E; Brückman, P; Brunet, J M; Bugge, L; Buschmann, P; Calvi, M; Camporesi, T; Canale, V; Carena, F; Castro, N; Cavallo, F R; Chapkin, M M; Charpentier, P; Checchia, P; Chierici, R; Shlyapnikov, P; Chudoba, J; Chung, S U; Cieslik, K; Collins, P; Contri, R; Cosme, G; Cossutti, F; Costa, M J; Crawley, B; Crennell, D J; Cuevas-Maestro, J; D'Hondt, J; Dalmau, J; Da Silva, T; Da Silva, W; Della Ricca, G; De Angelis, A; de Boer, Wim; De Clercq, C; De Lotto, B; De Maria, N; De Min, A; De Paula, L S; Di Ciaccio, Lucia; Di Simone, A; Doroba, K; Drees, J; Dris, M; Eigen, G; Ekelöf, T J C; Ellert, M; Elsing, M; Espirito-Santo, M C; Fanourakis, G K; Fassouliotis, D; Feindt, M; Fernández, J; Ferrer, A; Ferro, F; Flagmeyer, U; Föth, H; Fokitis, E; Fulda-Quenzer, F; Fuster, J A; Gandelman, M; García, C; Gavillet, P; Gazis, E N; Gokieli, R; Golob, B; Gómez-Ceballos, G; Gonçalves, P; Graziani, E; Grosdidier, G; Grzelak, K; Guy, J; Haag, C; Hallgren, A; Hamacher, K; Hamilton, K; Hansen, J; Haug, S; Hauler, F; Hedberg, V; Hennecke, M; Herr, H; Hoffman, J; Holmgren, S O; Holt, P J; Houlden, M A; Hultqvist, K; Jackson, J N; Jarlskog, G; Jarry, P; Jeans, D; Johansson, E K; Johansson, P D; Jonsson, P; Joram, C; Jungermann, L; Kapusta, F; Katsanevas, S; Katsoufis, E C; Kernel, G; Kersevan, Borut P; Kiiskinen, A P; King, B T; Kjaer, N J; Kluit, P; Kokkinias, P; Kourkoumelis, C; Kuznetsov, O; Krumshtein, Z; Kucharczyk, M; Lamsa, J; Leder, G; Ledroit, F; Leinonen, L; Leitner, R; Lemonne, J; Lepeltier, V; Lesiak, T; Liebig, W; Liko, D; Lipniacka, A; Lopes, J H; López, J M; Loukas, D; Lutz, P; Lyons, L; MacNaughton, J; Malek, A; Maltezos, S; Mandl, F; Marco, J; Marco, R; Maréchal, B; Margoni, M; Marin, J C; Mariotti, C; Markou, A; Martínez-Rivero, C; Masik, J; Mastroyiannopoulos, N; Matorras, F; Matteuzzi, C; Mazzucato, F; Mazzucato, M; McNulty, R; Meroni, C; Meyer, W T; Migliore, E; Mitaroff, W A; Mjörnmark, U; Moa, T; Moch, M; Mönig, K; Monge, R; Montenegro, J; Moraes, D; Moreno, S; Morettini, P; Müller, U; Münich, K; Mulders, M; Mundim, L M; Murray, W; Muryn, B; Myatt, Gerald; Myklebust, T; Nassiakou, M; Navarria, Francesco Luigi; Nawrocki, K; Nicolaidou, R; Nikolenko, M; Oblakowska-Mucha, A; Obraztsov, V F; Olshevskii, A G; Onofre, A; Orava, Risto; Österberg, K; Ouraou, A; Oyanguren, A; Paganoni, M; Paiano, S; Palacios, J P; Palka, H; Papadopoulou, T D; Pape, L; Parkes, C; Parodi, F; Parzefall, U; Passeri, A; Passon, O; Peralta, L; Perepelitsa, V F; Perrotta, A; Petrolini, A; Piedra, J; Pieri, L; Pierre, F; Pimenta, M; Piotto, E; Podobnik, T; Poireau, V; Pol, M E; Polok, G; Poropat, P; Pozdnyakov, V; Pukhaeva, N; Pullia, Antonio; Rames, J; Ramler, L; Read, A; Rebecchi, P; Rehn, J; Reid, D; Reinhardt, R; Renton, P B; Richard, F; Rídky, J; Rivero, M; Rodríguez, D; Romero, A; Ronchese, P; Rosenberg, E I; Roudeau, Patrick; Rovelli, T; Ruhlmann-Kleider, V; Ryabtchikov, D; Sadovskii, A; Salmi, L; Salt, J; Savoy-Navarro, A; Schwickerath, U; Segar, A; Sekulin, R L; Siebel, M; Sissakian, A N; Smadja, G; Smirnova, O G; Sokolov, A; Sopczak, A; Sosnowski, R; Spassoff, Tz; Stanitzki, M; Stocchi, A; Strauss, J; Stugu, B; Szczekowski, M; Szeptycka, M; Szumlak, T; Tabarelli de Fatis, T; Taffard, A C; Tegenfeldt, F; Timmermans, J; Tkatchev, L G; Tobin, M; Todorovova, S; Tomé, B; Tonazzo, A; Tortosa, P; Travnicek, P; Treille, D; Tristram, G; Trochimczuk, M; Troncon, C; Turluer, M L; Tyapkin, I A; Tyapkin, P; Tzamarias, S; Uvarov, V; Valenti, G; van Dam, P; Van Eldik, J; Van Lysebetten, A; Van Remortel, N; Van Vulpen, I B; Vegni, G; Veloso, F; Venus, W A; Verbeure, F; Verdier, P; Verzi, V; Vilanova, D; Vitale, L; Vrba, V; Wahlen, H; Washbrook, A J; Weiser, C; Wicke, D; Wickens, J H; Wilkinson, G; Winter, M; Witek, M; Yushchenko, O P; Zalewska-Bak, A; Zalewski, Piotr; Zavrtanik, D; Zhuravlov, V; Zimin, N I; Zinchenko, A I; Zupan, M

    2003-01-01

    eta_c(2980) production in gammagamma interactions has been detected via its decays into K0_sK+-pi-+, K+K-K+K- and K+K-pi+pi- in the data taken with the DELPHI detector at LEP1 and LEP2 energies. The two-photon radiative width averaged over all observed decay channels is Gamma_gammagamma = 13.9+-2.0(stat.)+-1.4(syst.)+-2.7(BR)keV. No direct decay channel eta_c -> pi+pi-pi+pi- has been observed. An upper limit Gamma_gammagamma<5.5keV at 95% confidence level has been evaluated for this decay mode.

  8. Results on two-photon interactions from Mark II at SPEAR

    Energy Technology Data Exchange (ETDEWEB)

    Abrams, G.S.; Alam, M.S.; Blocker, C.A.

    1979-10-01

    Preliminary results on two-photon interactions from the SLAC-LBL Mark II magnetic detector at SPEAR are presented. The cross section for eta' production by the reaction e/sup +/e/sup -/ ..-->.. e/sup +/e/sup -/ eta' has been measured over the beam energy range from 2 to 4 GeV. The radiative width GAMMA/sub ..gamma gamma../(eta') has been determined to be 5.8 +- 1.1 keV (+- 20% systematic uncertainty). Upper limits on the radiative widths of the f(1270), and A/sub 2/(1310) and f'(1515) mesons have been determined.

  9. Measurement of two-photon exchange effect by comparing elastic $e^\\pm p$ cross sections

    CERN Document Server

    Rimal, D; Raue, B A; Weinstein, L B; Arrington, J; Brooks, W K; Ungaro, M; Adhikari, K P; Akbar, Z; Pereira, S Anefalos; Badui, R A; Ball, J; Baltzell, N A; Battaglieri, M; Batourine, V; Bedlinskiy, I; Bennett, R P; Biselli, A S; Boiarinov, S; Briscoe, W J; Bültmann, S; Carman, D S; Celentano, A; Chetry, T; Ciullo, G; Clark, L; Colaneri, L; Cole, P L; Compton, N; Contalbrigo, M; Cortes, O; Crede, V; D'Angelo, A; Dashyan, N; De Vita, R; Deur, A; Djalali, C; Dupre, R; Egiyan, H; Alaoui, A El; Fassi, L El; Eugenio, P; Fedotov, G; Fersch, R; Filippi, A; Fleming, J A; Forest, T A; Fradi, A; Gevorgyan, N; Ghandilyan, Y; Gilfoyle, G P; Giovanetti, K L; Girod, F X; Gleason, C; Gohn, W; Golovatch, E; Gothe, R W; Griffioen, K A; Guo, L; Hafidi, K; Hanretty, C; Harrison, N; Hattawy, M; Heddle, D; Hicks, K; Holtrop, M; Hughes, S M; Ilieva, Y; Ireland, D G; Ishkhanov, B S; Isupov, E L; Jenkins, D; Jiang, H; Joosten, S; Keller, D; Khetarpal, P; Khachatryan, G; Khandaker, M; Kim, W; Klein, A; Klein, F J; Kubarovsky, V; Kuhn, S E; Kuleshov, S V; Lanza, L; Lenisa, P; Livingston, K; Lu, H Y; MacGregor, I J D; Markov, N; McKinnon, B; Mestayer, M D; Mirazita, M; Mokeev, V; Movsisyan, A; Munevar, E; Camacho, C Munoz; Nadel-Turonski, P; Ni, A; Niccolai, S; Niculescu, G; Niculescu, I; Osipenko, M; Ostrovidov, A I; Paolone, M; Paremuzyan, R; Park, K; Pasyuk, E; Phelps, W; Pisano, S; Pogorelko, O; Price, J W; Prok, Y; Protopopescu, D; Puckett, A J R; Rizzo, A; Rosner, G; Rossi, P; Roy, P; Sabatié, F; Salgado, C; Schumacher, R A; Seder, E; Sharabian, Y G; Skorodumina, Iu; Smith, G D; Sokhan, D; Sparveris, N; Stankovic, Ivana; Stepanyan, S; Strauch, S; Sytnik, V; Taiuti, M; Torayev, B; Voskanyan, H; Voutier, E; Walford, N K; Watts, D P; Wei, X; Wood, M H; Zachariou, N; Zana, L; Zhang, J; Zhao, Z W; Zonta, I

    2016-01-01

    [Background] The electromagnetic form factors of the proton measured by unpolarized and polarized electron scattering experiments show a significant disagreement that grows with the squared four momentum transfer ($Q^{2}$). Calculations have shown that the two measurements can be largely reconciled by accounting for the contributions of two-photon exchange (TPE). TPE effects are not typically included in the standard set of radiative corrections since theoretical calculations of the TPE effects are highly model dependent, and, until recently, no direct evidence of significant TPE effects has been observed. [Purpose] We measured the ratio of positron-proton to electron-proton elastic-scattering cross sections in order to determine the TPE contribution to elastic electron-proton scattering and thereby resolve the proton electric form factor discrepancy. [Methods] We produced a mixed simultaneous electron-positron beam in Jefferson Lab's Hall B by passing the 5.6 GeV primary electron beam through a radiator to p...

  10. Observation of the two-photon decay of a light penetrating particle

    Science.gov (United States)

    Faissner, H.; Frenzel, E.; Heinrigs, W.; Preussger, A.; Samm, D.; Samm, U.

    1981-07-01

    Coincident two-photon events, emerging from a 2 m long decay region, and pointing back to the SIN 590 MeV proton beam dump were detected in a thin-foil optical spark chamber. There was a significant excess of photons at small angles ( Eγ1 Eγ2 front of the decay region, but vanished with the wall put at its end. Presumably a light boson χ 0 comes from the beam dump, penetrates the shielding, and decays: χ0 → 2 γ The measured rate of (14.5 ± 5.0) events in 129 Coulomb fixes a combination of production cross section and life-time. If the new boson χ 0 was the axion, one can solve for the Higgs parameter X = 3.0 ± 0.3, and infer τa ≈ 7 ms, and ma = (250 ± 25) keV.

  11. 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.

  12. Two-Photon Polymerization lithography for three-dimensional micro polymer parts manufacturing evaluation

    DEFF Research Database (Denmark)

    Davoudinejad, Ali; Malureanu, Radu; Palima, Darwin

    2017-01-01

    Two-photon polymerization (2PP) technique is one of the common techniques to realize the fabrication of high-quality 3D microstructures. The combination between the laser power, the printing strategy, and the printed feature size are not completely assessed. This study characterizes the additive...... manufacturing processes by Direct Laser Writing (DLW) for fabrication of 3D microstructures. The printing samples were selected from a certified calibrated set with different sizes consisting of five boxes ranging from 8 μm to 200 μm. The laser power was selected as a variable parameter in order to find out...... for too low one. In addition, they show the importance of a good scaffolding, especially for bigger structures where the geometry can be distorted....

  13. Molecular beam resonant two-photon ionization study of caffeine and its hydrated clusters

    Science.gov (United States)

    Kim, Doory; Kim, Hyung Min; Yang, Key Young; Kim, Seong Keun; Kim, Nam Joon

    2008-04-01

    We investigated electronically excited states of caffeine and its 1:1 complex with water by using resonant two-photon ionization (R2PI) and UV-UV hole-burning techniques. Strong vibronic coupling between a pair of close-lying π-π * and n-π * transitions is proposed to be responsible for the broad spectral feature observed. By comparing the experimental results with those of theoretical calculations, both the O-bonded and N-bonded forms were suggested to be initially produced for the 1:1 complex. The electronic transitions of the O-bonded complex were blueshifted in the R2PI spectrum. For the N-bonded complex, the excited state undergoes an ultrafast decay process, followed by dissociation on a repulsive potential energy surface, which gives rise to a characteristically anomalous cluster distribution in nanosecond experiments.

  14. 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.

  15. Singlet oxygen production upon two-photon excitation of TiO2 in chloroform.

    Science.gov (United States)

    Li, Wenbing; Gandra, Naveen; Courtney, Shavelle N; Gao, Ruomei

    2009-08-03

    Singlet oxygen ((1)O(2)) signals observed upon illumination at 532 nm are a consequence of TiO(2) excitation in a two-photon process, and at 355 nm in a one-photon pathway. After corrections for light reflectance, the relative quantum yields of (1)O(2) production are 0.23+/-0.01 and 0.22+/-0.02 for 532 and 355 nm excitation, respectively. Total quenching rate constants of (1)O(2) removal by TiO(2) particles are near the diffusion-controlled rate limit. By employing (1)O(2) as a probe, we have established an efficient and simple method to elucidate the nature of TiO(2) photoexcitation.

  16. Excitonlike exchange in two-photon transitions of pairs of cold Rb Rydberg atoms

    Science.gov (United States)

    Lee, Jeonghun; Kongkhambut, Phatthamon; Gallagher, T. F.

    2017-12-01

    We have observed an excitonlike exchange in two-photon microwave transitions between pairs of cold Rb Rydberg atoms, specifically, transitions in which a n s1 /2n s1 /2 pair undergoes the transition to the n p1 /2n p3 /2 and n p3 /2n p1 /2 states. This transition occurs due to the excitonlike n s1 /2n pj↔n pjn s1 /2 exchange in the intermediate states, and the process can be thought of as a Förster resonant energy transfer between Floquet, or dressed, states. In addition, the measurements provide clear evidence of the importance of the three-dimensional nature of the dipole-dipole interaction.

  17. 3D high-resolution two-photon crosslinked hydrogel structures for biological studies.

    Science.gov (United States)

    Brigo, Laura; Urciuolo, Anna; Giulitti, Stefano; Della Giustina, Gioia; Tromayer, Maximilian; Liska, Robert; Elvassore, Nicola; Brusatin, Giovanna

    2017-06-01

    Hydrogels are widely used as matrices for cell growth due to the their tuneable chemical and physical properties, which mimic the extracellular matrix of natural tissue. The microfabrication of hydrogels into arbitrarily complex 3D structures is becoming essential for numerous biological applications, and in particular for investigating the correlation between cell shape and cell function in a 3D environment. Micrometric and sub-micrometric resolution hydrogel scaffolds are required to deeply investigate molecular mechanisms behind cell-matrix interaction and downstream cellular processes. We report the design and development of high resolution 3D gelatin hydrogel woodpile structures by two-photon crosslinking. Hydrated structures of lateral linewidth down to 0.5µm, lateral and axial resolution down to a few µm are demonstrated. According to the processing parameters, different degrees of polymerization are obtained, resulting in hydrated scaffolds of variable swelling and deformation. The 3D hydrogels are biocompatible and promote cell adhesion and migration. Interestingly, according to the polymerization degree, 3D hydrogel woodpile structures show variable extent of cell adhesion and invasion. Human BJ cell lines show capability of deforming 3D micrometric resolved hydrogel structures. The design and development of high resolution 3D gelatin hydrogel woodpile structures by two-photon crosslinking is reported. Significantly, topological and mechanical conditions of polymerized gelatin structures were suitable for cell accommodation in the volume of the woodpiles, leading to a cell density per unit area comparable to the bare substrate. The fabricated structures, presenting micrometric features of high resolution, are actively deformed by cells, both in terms of cell invasion within rods and of cell attachment in-between contiguous woodpiles. Possible biological targets for this 3D approach are customized 3D tissue models, or studies of cell adhesion

  18. Two-Photon Exchange Effects in Elastic Electron-Proton Scattering

    Energy Technology Data Exchange (ETDEWEB)

    Johnson, Myriam James [Argonne National Lab. (ANL), Argonne, IL (United States)

    2013-08-01

    Two methods, Rosenbluth separation and polarization transfer, can be used to extract the proton form factor ratio μp GEp/GMp, but they do not yield the same results. It is thought that the disagreement is due to two photon exchange corrections to the differential cross sections. High precision proton Rosenbluth extractions were carried out at 102 kinematics points spanning 16 values of momentum transfer Q2, from 0.40 to 5.76 GeV2. Reduced cross sections were found to 1.1% or better for Q2 less than 3 GeV2 increasing to 4% at 5.76 GeV2 The form factor ratios were determined to 1:5-3% for Q2 < 1.5 GeV2, increasing to 9% by 3 GeV2 and rapidly above. Our data agrees with prior Rosenbluth, improving upon it the 1.0 - 2.0 GeV2 range to conclusively show a separation from polarization transfer where it had not been certain before. In addition, reduced cross sections at each Q2 were tested for nonlinearity in the angular variable. Such a departure from linearity would be a signature of two photon exchange effects, and prior data had not been sufficiently precise to show nonzero curvature. Our data begins to hint at negative curvature but does not yet show a significant departure from zero.

  19. Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording and pharmacology

    Directory of Open Access Journals (Sweden)

    Manuel eLevy

    2012-12-01

    Full Text Available Uncovering the functional properties of individual synaptic inputs on single neurons is critical for understanding the computational role of synapses and dendrites. Previous studies combined whole-cell patch recording to load neurons with a fluorescent calcium indicator and two-photon imaging to map subcellular changes in fluorescence upon sensory stimulation. By hyperpolarizing the neuron below spike threshold, the patch electrode ensured that changes in fluorescence associated with synaptic events were isolated from those caused by back-propagating action potentials. This technique holds promise for determining whether the existence of unique cortical feature maps across different species may be associated with distinct wiring diagrams. However, the use of whole-cell patch for mapping inputs on dendrites is challenging in large mammals, due to brain pulsations and the accumulation of fluorescent dye in the extracellular milieu. Alternatively, sharp intracellular electrodes have been used to label neurons with fluorescent dyes, but the current passing capabilities of these high impedance electrodes may be insufficient to prevent spiking. In this study, we tested whether sharp electrode recording is suitable for mapping functional inputs on dendrites in the cat visual cortex. We compared three different strategies for suppressing visually evoked spikes: (1 hyperpolarization by intracellular current injection, (2 pharmacological blockade of voltage-gated sodium channels by intracellular QX-314, and (3 GABA iontophoresis from a perisomatic electrode glued to the intracellular electrode. We found that functional inputs on dendrites could be successfully imaged using all three strategies. However, the best method for preventing spikes was GABA iontophoresis with low currents (5 to 10 nA, which minimally affected the local circuit. Our methods advance the possibility of determining functional connectivity in preparations where whole-cell patch may be

  20. The Use of Two-Photon FRET-FLIM to Study Protein Interactions During Nuclear Envelope Fusion In Vivo and In Vitro.

    Science.gov (United States)

    Byrne, Richard D; Larijani, Banafshé; Poccia, Dominic L

    2016-01-01

    FRET-FLIM techniques have wide application in the study of protein and protein-lipid interactions in cells. We have pioneered an imaging platform for accurate detection of functional states of proteins and their interactions in fixed cells. This platform, two-site-amplified Förster resonance energy transfer (a-FRET), allows greater signal generation while retaining minimal noise thus enabling application of fluorescence lifetime imaging microscopy (FLIM) to be routinely deployed in different types of cells and tissue. We have used the method described here, time-resolved FRET monitored by two-photon FLIM, to demonstrate the direct interaction of Phospholipase Cγ (PLCγ) by Src Family Kinase 1 (SFK1) during nuclear envelope formation and during male and female pronuclear membrane fusion in fertilized sea urchin eggs. We describe here a generic method that can be applied to monitor any proteins of interest.