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

Science.gov (United States)

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

2018-02-01

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

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

Science.gov (United States)

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

2017-04-01

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

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

Science.gov (United States)

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

2018-05-16

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

Human bladder cancer diagnosis using multiphoton microscopy

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

2009-02-01

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

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

Science.gov (United States)

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

2012-08-01

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

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

OpenAIRE

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

2016-01-01

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

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

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

2010-03-01

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

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

Science.gov (United States)

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

2017-07-01

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

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

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

2012-01-01

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

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

International Nuclear Information System (INIS)

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

2014-01-01

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

Coherent beam control through inhomogeneous media in multi-photon microscopy

Science.gov (United States)

Paudel, Hari Prasad

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

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

International Nuclear Information System (INIS)

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

2016-01-01

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

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

Science.gov (United States)

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

2016-01-01

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

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

OpenAIRE

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

2012-01-01

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

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

Science.gov (United States)

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

2016-05-01

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

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

Science.gov (United States)

Salama, Paul

2008-02-01

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

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

Science.gov (United States)

Lai, Zhenhua

The author's work is divided into three aspects: multimodal microscopy, stepwise multi-photon activation fluorescence (SMPAF) of melanin, and customized-profile lenses (CPL) for on-axis laser scanners, which will be introduced respectively. A multimodal microscope provides the ability to image samples with multiple modalities on the same stage, which incorporates the benefits of all modalities. The multimodal microscopes developed in this dissertation are the Keck 3D fusion multimodal microscope 2.0 (3DFM 2.0), upgraded from the old 3DFM with improved performance and flexibility, and the multimodal microscope for targeting small particles (the "Target" system). The control systems developed for both microscopes are low-cost and easy-to-build, with all components off-the-shelf. The control system have not only significantly decreased the complexity and size of the microscope, but also increased the pixel resolution and flexibility. The SMPAF of melanin, activated by a continuous-wave (CW) mode near-infrared (NIR) laser, has potential applications for a low-cost and reliable method of detecting melanin. The photophysics of melanin SMPAF has been studied by theoretical analysis of the excitation process and investigation of the spectra, activation threshold, and photon number absorption of melanin SMPAF. SMPAF images of melanin in mouse hair and skin, mouse melanoma, and human black and white hairs are compared with images taken by conventional multi-photon fluorescence microscopy (MPFM) and confocal reflectance microscopy (CRM). SMPAF images significantly increase specificity and demonstrate the potential to increase sensitivity for melanin detection compared to MPFM images and CRM images. Employing melanin SMPAF imaging to detect melanin inside human skin in vivo has been demonstrated, which proves the effectiveness of melanin detection using SMPAF for medical purposes. Selective melanin ablation with micrometer resolution has been presented using the Target system

Visualizing liver anatomy, physiology and pharmacology using multiphoton microscopy.

Science.gov (United States)

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

2017-01-01

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

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

Science.gov (United States)

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

2016-05-01

We report on a miniature fiber-optic multiphoton microscopy (MPM) system based on a frequency-doubled femtosecond Er-doped fiber laser. The femtosecond pulses from the laser source are delivered to the miniature fiber-optic probe at 1.58 µm wavelength, where a standard single mode fiber is used for delivery without the need of free-space dispersion compensation components. The beam is frequency-doubled inside the probe by a periodically poled MgO:LiNbO3 crystal. Frequency-doubled pulses at 786 nm with a maximum power of 80 mW and a pulsewidth of 150 fs are obtained and applied to excite intrinsic signals from tissues. A MEMS scanner, a miniature objective, and a multimode collection fiber are further used to make the probe compact. The miniature fiber-optic MPM system is highly portable and robust. Ex vivo multiphoton imaging of mammalian skins demonstrates the capability of the system in imaging biological tissues. The results show that the miniature fiber-optic MPM system using frequency-doubled femtosecond fiber laser can potentially bring the MPM imaging for clinical applications.

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

Science.gov (United States)

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

2012-08-13

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

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

Science.gov (United States)

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

2015-02-01

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

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

Science.gov (United States)

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

2012-02-01

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

Current developments in clinical multiphoton tomography

Science.gov (United States)

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

2010-02-01

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

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

Science.gov (United States)

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

2017-01-01

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

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

Directory of Open Access Journals (Sweden)

Hyun Mi Ju

2017-07-01

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

Identification of intramural metastasis in esophageal cancer using multiphoton microscopy

Science.gov (United States)

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

2017-02-01

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

Full optical model of micro-endoscope with optical coherence microscopy, multiphoton microscopy and visible capabilities

Science.gov (United States)

Vega, David; Kiekens, Kelli C.; Syson, Nikolas C.; Romano, Gabriella; Baker, Tressa; Barton, Jennifer K.

2018-02-01

While Optical Coherence Microscopy (OCM), Multiphoton Microscopy (MPM), and narrowband imaging are powerful imaging techniques that can be used to detect cancer, each imaging technique has limitations when used by itself. Combining them into an endoscope to work in synergy can help achieve high sensitivity and specificity for diagnosis at the point of care. Such complex endoscopes have an elevated risk of failure, and performing proper modelling ensures functionality and minimizes risk. We present full 2D and 3D models of a multimodality optical micro-endoscope to provide real-time detection of carcinomas, called a salpingoscope. The models evaluate the endoscope illumination and light collection capabilities of various modalities. The design features two optical paths with different numerical apertures (NA) through a single lens system with a scanning optical fiber. The dual path is achieved using dichroic coatings embedded in a triplet. A high NA optical path is designed to perform OCM and MPM while a low NA optical path is designed for the visible spectrum to navigate the endoscope to areas of interest and narrowband imaging. Different tests such as the reflectance profile of homogeneous epithelial tissue were performed to adjust the models properly. Light collection models for the different modalities were created and tested for efficiency. While it is challenging to evaluate the efficiency of multimodality endoscopes, the models ensure that the system is design for the expected light collection levels to provide detectable signal to work for the intended imaging.

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

Science.gov (United States)

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

2012-12-01

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

A pragmatic guide to multiphoton microscope design

Science.gov (United States)

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

2016-01-01

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

Non-invasive in vivo characterization of skin wound healing using label-free multiphoton microscopy (Conference Presentation)

Science.gov (United States)

Jones, Jake D.; Majid, Fariah; Ramser, Hallie; Quinn, Kyle P.

2017-02-01

Non-healing ulcerative wounds, such as diabetic foot ulcers, are challenging to diagnose and treat due to their numerous possible etiologies and the variable efficacy of advanced wound care products. Thus, there is a critical need to develop new quantitative biomarkers and diagnostic technologies that are sensitive to wound status in order to guide care. The objective of this study was to evaluate the utility of label-free multiphoton microscopy for characterizing wound healing dynamics in vivo and identifying potential differences in diabetic wounds. We isolated and measured an optical redox ratio of FAD/(NADH+FAD) autofluorescence to provide three-dimensional maps of local cellular metabolism. Using a mouse model of wound healing, in vivo imaging at the wound edge identified a significant decrease in the optical redox ratio of the epidermis (p≤0.0103) between Days 3 through 14 compared to Day 1. This decrease in redox ratio coincided with a decrease in NADH fluorescence lifetime and thickening of the epithelium, collectively suggesting a sensitivity to keratinocyte hyperproliferation. In contrast to normal wounds, we have found that keratinocytes from diabetic wounds remain in a proliferative state at later time points with a lower redox ratio at the wound edge. Microstructural organization and composition was also measured from second harmonic generation imaging of collagen and revealed differences between diabetic and non-diabetic wounds. Our work demonstrates label-free multiphoton microscopy offers potential to provide non-invasive structural and functional biomarkers associated with different stages of skin wound healing, which may be used to detect delayed healing and guide treatment.

Multi-photon microscope driven by novel green laser pump

DEFF Research Database (Denmark)

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

2016-01-01

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

Characteristics of subgingival calculus detection by multiphoton fluorescence microscopy

Science.gov (United States)

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

2011-06-01

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

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

DEFF Research Database (Denmark)

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

2013-01-01

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

Proximal design for a multimodality endoscope with multiphoton microscopy, optical coherence microscopy and visual modalities

Science.gov (United States)

Kiekens, Kelli C.; Talarico, Olivia; Barton, Jennifer K.

2018-02-01

A multimodality endoscope system has been designed for early detection of ovarian cancer. Multiple illumination and detection systems must be integrated in a compact, stable, transportable configuration to meet the requirements of a clinical setting. The proximal configuration presented here supports visible light navigation with a large field of view and low resolution, high resolution multiphoton microscopy (MPM), and high resolution optical coherence microscopy (OCM). All modalities are integrated into a single optical system in the endoscope. The system requires two light sources: a green laser for visible light navigation and a compact fiber based femtosecond laser for MPM and OCM. Using an inline wavelength division multiplexer, the two sources are combined into a single mode fiber. To accomplish OCM, a fiber coupler is used to separate the femtosecond laser into a reference arm and signal arm. The reflected reference arm and the signal from the sample are interfered and wavelength separated by a reflection grating and detected using a linear array. The MPM signal is collimated and goes through a series of filters to separate the 2nd and 3rd harmonics as well as twophoton excitation florescence (2PEF) and 3PEF. Each signal is independently detected on a photo multiplier tube and amplified. The visible light is collected by multiple high numerical aperture fibers at the endoscope tip which are bundled into one SMA adapter at the proximal end and connected to a photodetector. This integrated system design is compact, efficient and meets both optical and mechanical requirements for clinical applications.

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

DEFF Research Database (Denmark)

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

2015-01-01

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

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

Science.gov (United States)

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

2004-07-01

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

Identifying Two Common Types of Breast Benign Diseases Based on Multiphoton Microscopy

Directory of Open Access Journals (Sweden)

Yan Wu

2018-01-01

Full Text Available Multiphoton microscopy has attracted increasing attention and investigations in the field of breast cancer, based on two-photon excited fluorescence (TPEF and second-harmonic generation (SHG. However, the incidence of breast benign diseases is about 5 to 10 times higher than breast cancer; up to 30% of women suffer from breast benign diseases and require treatment at some time in their lives. Thus, in this study, MPM was applied to image fibroadenoma and fibrocystic lesion, which are two of the most common breast benign diseases. The results show that MPM has the capability to identify the microstructure of lobule and stroma in normal breast tissue, the interaction of compressed ducts with surrounding collagen fiber in fibroadenoma, and the architecture of cysts filled with cystic fluid in fibrocystic disease. These findings indicate that, with integration of MPM into currently accepted clinical imaging system, it has the potential to make a real-time diagnosis of breast benign diseases in vivo, as well as breast cancer.

Electronically tunable femtosecond all-fiber optical parametric oscillator for multi-photon microscopy

Science.gov (United States)

Hellwig, Tim; Brinkmann, Maximilian; Fallnich, Carsten

2018-02-01

We present a femtosecond fiber-based optical parametric oscillator (FOPO) for multiphoton microscopy with wavelength tuning by electronic repetition rate tuning in combination with a dispersive filter in the FOPO cavity. The all-spliced, all-fiber FOPO cavity is based on polarization-maintaining fibers and a broadband output coupler, allowing to get access to the resonant signal pulses as well as the idler pulses simultaneously. The system was pumped by a gain-switched fiber-coupled laser diode emitting pulses at a central wavelength of 1030 nm and an electronically tunable repetition frequency of about 2 MHz. The pump pulses were amplified in an Ytterbium fiber amplifier system with a pulse duration after amplification of 13 ps. Tuning of the idler (1140 nm - 1300 nm) and signal wavelengths (850 nm - 940 nm) was achieved by changing the repetition frequency of the pump laser by about 4 kHz. The generated signal pulses reached a pulse energy of up to 9.2 nJ at 920 nm and were spectrally broadened to about 6 nm in the FOPO by a combination of self-phase and cross-phase modulation. We showed external compression of the idler pulses at 920 nm to about 430 fs and appleid them to two-photon excitation microscopy with green fluorescent dyes. The presented system constitutes an important step towards a fully fiber-integrated all-electronically tunable and, thereby, programmable light source and already embodies a versatile and flexible light source for applications, e.g., for smart microscopy.

Label-free detection of breast masses using multiphoton microscopy.

Directory of Open Access Journals (Sweden)

Xiufeng Wu

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

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

Energy Technology Data Exchange (ETDEWEB)

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

2015-12-07

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

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

International Nuclear Information System (INIS)

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

2015-01-01

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

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

International Nuclear Information System (INIS)

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

2013-01-01

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

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

DEFF Research Database (Denmark)

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

2011-01-01

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

Non-descanned multifocal multiphoton microscopy with a multianode photomultiplier tube

International Nuclear Information System (INIS)

Cha, Jae Won; Yew, Elijah Y. S.; Kim, Daekeun; Subramanian, Jaichandar; Nedivi, Elly; So, Peter T. C.

2015-01-01

Multifocal multiphoton microscopy (MMM) improves imaging speed over a point scanning approach by parallelizing the excitation process. Early versions of MMM relied on imaging detectors to record emission signals from multiple foci simultaneously. For many turbid biological specimens, the scattering of emission photons results in blurred images and degrades the signal-to-noise ratio (SNR). We have recently demonstrated that a multianode photomultiplier tube (MAPMT) placed in a descanned configuration can effectively collect scattered emission photons from each focus into their corresponding anodes significantly improving image SNR for highly scattering specimens. Unfortunately, a descanned MMM has a longer detection path resulting in substantial emission photon loss. Optical design constraints in a descanned geometry further results in significant optical aberrations especially for large field-of-view (FOV), high NA objectives. Here, we introduce a non-descanned MMM based on MAPMT that substantially overcomes most of these drawbacks. We show that we improve signal efficiency up to fourfold with limited image SNR degradation due to scattered emission photons. The excitation foci can also be spaced wider to cover the full FOV of the objective with minimal aberrations. The performance of this system is demonstrated by imaging interneuron morphological structures deep in the brains of living mice

Non-descanned multifocal multiphoton microscopy with a multianode photomultiplier tube

Energy Technology Data Exchange (ETDEWEB)

Cha, Jae Won; Yew, Elijah Y. S. [Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA (United States); Kim, Daekeun [Department of Mechanical Engineering, Dankook University (Korea, Republic of); Subramanian, Jaichandar [Picower Institute for Learning and Memory, Massachusetts Institute of Technology, Cambridge, MA (United States); Nedivi, Elly [Picower Institute for Learning and Memory, Massachusetts Institute of Technology, Cambridge, MA (United States); Departments of Brain and Cognitive Sciences, and Biology, Massachusetts Institute of Technology, Cambridge, MA (United States); So, Peter T. C. [Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA (United States); Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA (United States)

2015-08-15

Multifocal multiphoton microscopy (MMM) improves imaging speed over a point scanning approach by parallelizing the excitation process. Early versions of MMM relied on imaging detectors to record emission signals from multiple foci simultaneously. For many turbid biological specimens, the scattering of emission photons results in blurred images and degrades the signal-to-noise ratio (SNR). We have recently demonstrated that a multianode photomultiplier tube (MAPMT) placed in a descanned configuration can effectively collect scattered emission photons from each focus into their corresponding anodes significantly improving image SNR for highly scattering specimens. Unfortunately, a descanned MMM has a longer detection path resulting in substantial emission photon loss. Optical design constraints in a descanned geometry further results in significant optical aberrations especially for large field-of-view (FOV), high NA objectives. Here, we introduce a non-descanned MMM based on MAPMT that substantially overcomes most of these drawbacks. We show that we improve signal efficiency up to fourfold with limited image SNR degradation due to scattered emission photons. The excitation foci can also be spaced wider to cover the full FOV of the objective with minimal aberrations. The performance of this system is demonstrated by imaging interneuron morphological structures deep in the brains of living mice.

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

Science.gov (United States)

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

2008-02-01

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

Visualizing Angiogenesis by Multiphoton Microscopy In Vivo in Genetically Modified 3D-PLGA/nHAp Scaffold for Calvarial Critical Bone Defect Repair.

Science.gov (United States)

Li, Jian; Jahr, Holger; Zheng, Wei; Ren, Pei-Gen

2017-09-07

The reconstruction of critically sized bone defects remains a serious clinical problem because of poor angiogenesis within tissue-engineered scaffolds during repair, which gives rise to a lack of sufficient blood supply and causes necrosis of the new tissues. Rapid vascularization is a vital prerequisite for new tissue survival and integration with existing host tissue. The de novo generation of vasculature in scaffolds is one of the most important steps in making bone regeneration more efficient, allowing repairing tissue to grow into a scaffold. To tackle this problem, the genetic modification of a biomaterial scaffold is used to accelerate angiogenesis and osteogenesis. However, visualizing and tracking in vivo blood vessel formation in real-time and in three-dimensional (3D) scaffolds or new bone tissue is still an obstacle for bone tissue engineering. Multiphoton microscopy (MPM) is a novel bio-imaging modality that can acquire volumetric data from biological structures in a high-resolution and minimally-invasive manner. The objective of this study was to visualize angiogenesis with multiphoton microscopy in vivo in a genetically modified 3D-PLGA/nHAp scaffold for calvarial critical bone defect repair. PLGA/nHAp scaffolds were functionalized for the sustained delivery of a growth factor pdgf-b gene carrying lentiviral vectors (LV-pdgfb) in order to facilitate angiogenesis and to enhance bone regeneration. In a scaffold-implanted calvarial critical bone defect mouse model, the blood vessel areas (BVAs) in PHp scaffolds were significantly higher than in PH scaffolds. Additionally, the expression of pdgf-b and angiogenesis-related genes, vWF and VEGFR2, increased correspondingly. MicroCT analysis indicated that the new bone formation in the PHp group dramatically improved compared to the other groups. To our knowledge, this is the first time multiphoton microscopy was used in bone tissue-engineering to investigate angiogenesis in a 3D bio-degradable scaffold in

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

Energy Technology Data Exchange (ETDEWEB)

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

2007-08-21

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

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

Science.gov (United States)

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

2014-09-01

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

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

Science.gov (United States)

Puschmann, Stefan; Rahn, Christian-Dennis; Wenck, Horst; Gallinat, Stefan; Fischer, Frank

2012-03-01

Extracellular skin structures in human skin are impaired during intrinsic and extrinsic aging. Assessment of these dermal changes is conducted by subjective clinical evaluation and histological and molecular analysis. We aimed to develop a new parameter for the noninvasive quantitative determination of dermal skin alterations utilizing the high-resolution three-dimensional multiphoton laser scanning microscopy (MPLSM) technique. To quantify structural differences between chronically sun-exposed and sun-protected human skin, the respective collagen-specific second harmonic generation and the elastin-specific autofluorescence signals were recorded in young and elderly volunteers using the MPLSM technique. After image processing, the elastin-to-collagen ratio (ELCOR) was calculated. Results show that the ELCOR parameter of volar forearm skin significantly increases with age. For elderly volunteers, the ELCOR value calculated for the chronically sun-exposed temple area is significantly augmented compared to the sun-protected upper arm area. Based on the MPLSM technology, we introduce the ELCOR parameter as a new means to quantify accurately age-associated alterations in the extracellular matrix.

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

Energy Technology Data Exchange (ETDEWEB)

Zhuo, Shuangmu, E-mail: shuangmuzhuo@gmail.com, E-mail: hanry-yu@nuhs.edu.sg [Biosystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #04-13/14 Enterprise Wing, 138602 Singapore (Singapore); Institute of Laser and Optoelectronics Technology, Fujian Normal University, Fuzhou 350007 (China); Yan, Jie [Biosystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #04-13/14 Enterprise Wing, 138602 Singapore (Singapore); Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, #04-01, 138669 Singapore (Singapore); Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, 14 Medical Drive, MD 11 #04-01A, 117599 Singapore (Singapore); Kang, Yuzhan [Biosystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #04-13/14 Enterprise Wing, 138602 Singapore (Singapore); Xu, Shuoyu [Biosystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #04-13/14 Enterprise Wing, 138602 Singapore (Singapore); Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, #04-01, 138669 Singapore (Singapore); Computation and System Biology Program, Singapore-MIT Alliance, 4 Engineering Drive 3, E4-04-10, 117576 Singapore (Singapore); Peng, Qiwen [Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, #04-01, 138669 Singapore (Singapore); Computation and System Biology Program, Singapore-MIT Alliance, 4 Engineering Drive 3, E4-04-10, 117576 Singapore (Singapore); Mechanobiology Institute, 5A Engineering Drive 1, T-Lab #05-01, 117411 Singapore (Singapore); and others

2014-07-14

Various structural features on the liver surface reflect functional changes in the liver. The visualization of these surface features with molecular specificity is of particular relevance to understanding the physiology and diseases of the liver. Using multi-photon microscopy (MPM), we have developed a label-free, three-dimensional quantitative and sensitive method to visualize various structural features of liver surface in living rat. MPM could quantitatively image the microstructural features of liver surface with respect to the sinuosity of collagen fiber, the elastic fiber structure, the ratio between elastin and collagen, collagen content, and the metabolic state of the hepatocytes that are correlative with the pathophysiologically induced changes in the regions of interest. This study highlights the potential of this technique as a useful tool for pathophysiological studies and possible diagnosis of the liver diseases with further development.

Multi-photon microscopy with a low-cost and highly efficient Cr:LiCAF laser

Science.gov (United States)

Sakadić, Sava; Demirbas, Umit; Mempel, Thorsten R.; Moore, Anna; Ruvinskaya, Svetlana; Boas, David A.; Sennaroglu, Alphan; Kartner, Franz X.; Fujimoto, James G.

2009-01-01

Multi-photon microscopy (MPM) is a powerful tool for biomedical imaging, enabling molecular contrast and integrated structural and functional imaging on the cellular and subcellular level. However, the cost and complexity of femtosecond laser sources that are required in MPM are significant hurdles to widespread adoption of this important imaging modality. In this work, we describe femtosecond diode pumped Cr:LiCAF laser technology as a low cost alternative to femtosecond Ti:Sapphire lasers for MPM. Using single mode pump diodes which cost only $150 each, a diode pumped Cr:LiCAF laser generates ~70-fs duration, 1.8-nJ pulses at ~800 nm wavelengths, with a repetition rate of 100 MHz and average output power of 180 mW. Representative examples of MPM imaging in neuroscience, immunology, endocrinology and cancer research using Cr:LiCAF laser technology are presented. These studies demonstrate the potential of this laser source for use in a broad range of MPM applications. PMID:19065223

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

Science.gov (United States)

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

2009-08-01

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

Multiphoton processes

International Nuclear Information System (INIS)

Manus, C.; Mainfray, G.

1980-01-01

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

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

Energy Technology Data Exchange (ETDEWEB)

Darvin, M E; Richter, H; Zhu, Y J; Meinke, M C; Knorr, F; Lademann, J [Center of Experimental and Applied Cutaneous Physiology, Department of Dermatology, Venerology and Allergology, Charité - Universitätsmedizin Berlin (Germany); Gonchukov, S A [National Research Nuclear University ' ' MEPhI' ' (Russian Federation); Koenig, K [JenLab GmbH, Schillerstr. 1, 07745 Jena (Germany)

2014-07-31

Two state-of-the-art microscopic optical methods, namely, confocal laser scanning microscopy in the fluorescence and reflectance regimes and multiphoton tomography in the autofluorescence and second harmonic generation regimes, are compared for porcine skin ex vivo and healthy human skin in vivo. All skin layers such as stratum corneum (SC), stratum spinosum (SS), stratum basale (SB), papillary dermis (PD) and reticular dermis (RD) as well as transition zones between these skin layers are measured noninvasively at a high resolution, using the above mentioned microscopic methods. In the case of confocal laser scanning microscopy (CLSM), measurements in the fluorescence regime were performed by using a fluorescent dye whose topical application on the surface is well suited for the investigation of superficial SC and characterisation of the skin barrier function. For investigations of deeply located skin layers, such as SS, SB and PD, the fluorescent dye must be injected into the skin, which markedly limits fluorescence measurements using CLSM. In the case of reflection CLSM measurements, the obtained results can be compared to the results of multiphoton tomography (MPT) for all skin layers excluding RD. CLSM cannot distinguish between dermal collagen and elastin measuring their superposition in the RD. By using MPT, it is possible to analyse the collagen and elastin structures separately, which is important for the investigation of anti-aging processes. The resolution of MPT is superior to CLSM. The advantages and limitations of both methods are discussed and the differences and similarities between human and porcine skin are highlighted. (laser biophotonics)

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

Science.gov (United States)

Darvin, M. E.; Richter, H.; Zhu, Y. J.; Meinke, M. C.; Knorr, F.; Gonchukov, S. A.; Koenig, K.; Lademann, J.

2014-07-01

Two state-of-the-art microscopic optical methods, namely, confocal laser scanning microscopy in the fluorescence and reflectance regimes and multiphoton tomography in the autofluorescence and second harmonic generation regimes, are compared for porcine skin ex vivo and healthy human skin in vivo. All skin layers such as stratum corneum (SC), stratum spinosum (SS), stratum basale (SB), papillary dermis (PD) and reticular dermis (RD) as well as transition zones between these skin layers are measured noninvasively at a high resolution, using the above mentioned microscopic methods. In the case of confocal laser scanning microscopy (CLSM), measurements in the fluorescence regime were performed by using a fluorescent dye whose topical application on the surface is well suited for the investigation of superficial SC and characterisation of the skin barrier function. For investigations of deeply located skin layers, such as SS, SB and PD, the fluorescent dye must be injected into the skin, which markedly limits fluorescence measurements using CLSM. In the case of reflection CLSM measurements, the obtained results can be compared to the results of multiphoton tomography (MPT) for all skin layers excluding RD. CLSM cannot distinguish between dermal collagen and elastin measuring their superposition in the RD. By using MPT, it is possible to analyse the collagen and elastin structures separately, which is important for the investigation of anti-aging processes. The resolution of MPT is superior to CLSM. The advantages and limitations of both methods are discussed and the differences and similarities between human and porcine skin are highlighted.

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

International Nuclear Information System (INIS)

Darvin, M E; Richter, H; Zhu, Y J; Meinke, M C; Knorr, F; Lademann, J; Gonchukov, S A; Koenig, K

2014-01-01

Two state-of-the-art microscopic optical methods, namely, confocal laser scanning microscopy in the fluorescence and reflectance regimes and multiphoton tomography in the autofluorescence and second harmonic generation regimes, are compared for porcine skin ex vivo and healthy human skin in vivo. All skin layers such as stratum corneum (SC), stratum spinosum (SS), stratum basale (SB), papillary dermis (PD) and reticular dermis (RD) as well as transition zones between these skin layers are measured noninvasively at a high resolution, using the above mentioned microscopic methods. In the case of confocal laser scanning microscopy (CLSM), measurements in the fluorescence regime were performed by using a fluorescent dye whose topical application on the surface is well suited for the investigation of superficial SC and characterisation of the skin barrier function. For investigations of deeply located skin layers, such as SS, SB and PD, the fluorescent dye must be injected into the skin, which markedly limits fluorescence measurements using CLSM. In the case of reflection CLSM measurements, the obtained results can be compared to the results of multiphoton tomography (MPT) for all skin layers excluding RD. CLSM cannot distinguish between dermal collagen and elastin measuring their superposition in the RD. By using MPT, it is possible to analyse the collagen and elastin structures separately, which is important for the investigation of anti-aging processes. The resolution of MPT is superior to CLSM. The advantages and limitations of both methods are discussed and the differences and similarities between human and porcine skin are highlighted. (laser biophotonics)

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

Science.gov (United States)

Abraham, Thomas; Kayra, Damian; Zhang, Angela; Suzuki, Masaru; McDonough, John; Elliott, W. M.; Cooper, Joel D.; Hogg, James C.

2013-02-01

Lung is a complex gas exchanger with interfacial area (where the gas exchange takes place) is about the size of a tennis court. Respiratory function is linked to the biomechanical stability of the gas exchange or alveolar regions which directly depends on the spatial distributions of the extracellular matrix fibers such fibrillar collagens and elastin fibers. It is very important to visualize and quantify these fibers at their native and inflated conditions to have correct morphometric information on differences between control and diseased states. This can be only achieved in the ex vivo states by imaging directly frozen lung specimens inflated to total lung capacity. Multiphoton microscopy, which uses ultra-short infrared laser pulses as the excitation source, produces multiphoton excitation fluorescence (MPEF) signals from endogenously fluorescent proteins (e.g. elastin) and induces specific second harmonic generation (SHG) signals from non-centrosymmetric proteins such as fibrillar collagens in fresh human lung tissues [J. Struct. Biol. (2010)171,189-196]. Here we report for the first time 3D image data obtained directly from thick frozen inflated lung specimens (~0.7- 1.0 millimeter thick) visualized at -60°C without prior fixation or staining in healthy and diseased states. Lung specimens donated for transplantation and released for research when no appropriate recipient was identified served as controls, and diseased lung specimens donated for research by patients receiving lung transplantation for very severe COPD (n=4) were prepared as previously described [N. Engl. J. Med. (2011) 201, 1567]. Lung slices evenly spaced between apex and base were examined using multiphoton microscopy while maintained at -60°C using a temperature controlled cold stage with a temperature resolution of 0.1°C. Infrared femto-second laser pulses tuned to 880nm, dry microscopic objectives, and non-de-scanned detectors/spectrophotometer located in the reflection geometry were

Stepwise multiphoton activation fluorescence reveals a new method of melanin detection

Science.gov (United States)

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

2013-06-01

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

Multiphoton processes: conference proceedings

International Nuclear Information System (INIS)

Lambropoulos, P.; Smith, S.J.

1984-01-01

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

Multiphoton processes: conference proceedings

Energy Technology Data Exchange (ETDEWEB)

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

1984-01-01

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

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

Science.gov (United States)

Wilson, Jesse W; Park, Jong Kang; Warren, Warren S; Fischer, Martin C

2015-03-01

The lock-in amplifier is a critical component in many different types of experiments, because of its ability to reduce spurious or environmental noise components by restricting detection to a single frequency and phase. One example application is pump-probe microscopy, a multiphoton technique that leverages excited-state dynamics for imaging contrast. With this application in mind, we present here the design and implementation of a high-speed lock-in amplifier on the field-programmable gate array (FPGA) coprocessor of a data acquisition board. The most important advantage is the inherent ability to filter signals based on more complex modulation patterns. As an example, we use the flexibility of the FPGA approach to enable a novel pump-probe detection scheme based on spread-spectrum communications techniques.

MULTI-PHOTON PHOSPHOR FEASIBILITY RESEARCH

Energy Technology Data Exchange (ETDEWEB)

R. Graham; W. Chow

2003-05-01

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

Raman Microscopy: A Noninvasive Method to Visualize the Localizations of Biomolecules in the Cornea.

Science.gov (United States)

Kaji, Yuichi; Akiyama, Toshihiro; Segawa, Hiroki; Oshika, Tetsuro; Kano, Hideaki

2017-11-01

In vivo and in situ visualization of biomolecules without pretreatment will be important for diagnosis and treatment of ocular disorders in the future. Recently, multiphoton microscopy, based on the nonlinear interactions between molecules and photons, has been applied to reveal the localizations of various molecules in tissues. We aimed to use multimodal multiphoton microscopy to visualize the localizations of specific biomolecules in rat corneas. Multiphoton images of the corneas were obtained from nonlinear signals of coherent anti-Stokes Raman scattering, third-order sum frequency generation, and second-harmonic generation. The localizations of the adhesion complex-containing basement membrane and Bowman layer were clearly visible in the third-order sum frequency generation images. The fine structure of type I collagen was observed in the corneal stroma in the second-harmonic generation images. The localizations of lipids, proteins, and nucleic acids (DNA/RNA) was obtained in the coherent anti-Stokes Raman scattering images. Imaging technologies have progressed significantly and been applied in medical fields. Optical coherence tomography and confocal microscopy are widely used but do not provide information on the molecular structure of the cornea. By contrast, multiphoton microscopy provides information on the molecular structure of living tissues. Using this technique, we successfully visualized the localizations of various biomolecules including lipids, proteins, and nucleic acids in the cornea. We speculate that multiphoton microscopy will provide essential information on the physiological and pathological conditions of the cornea, as well as molecular localizations in tissues without pretreatment.

Identification of tumor cells infiltrating into connective tissue in esophageal cancer by multiphoton microscopy

Science.gov (United States)

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

2016-10-01

Esophageal cancer is one of the most common malignancies of the gastrointestinal cancers and carries poorer prognosis than other gastrointestinal cancers. In general practice, the depth of tumor infiltration in esophageal wall is crucial to establishing appropriate treatment plan which is established by detecting the tumor infiltration depth. Connective tissue is one of the main structures that form the esophageal wall. So, identification of tumor cells infiltrating into connective tissue is helping for detecting the tumor infiltration depth. Our aim is to evaluate whether multiphoton microscopy (MPM) can be used to detect tumor cells infiltrating into connective tissue in the esophageal cancer. MPM is well-suited for real-time detecting morphologic and cellular changes in fresh tissues since many endogenous fluorophores of fresh tissues are excited through two-photon excited fluorescence (TPEF) and second harmonic generation (SHG). In this work, microstructure of tumor cells and connective tissue are first studied. Then, morphological changes of collagen fibers after the infiltration of tumor cells are shown. These results show that MPM has the ability to detect tumor cells infiltrating into connective tissue in the esophageal cancer. In the future, MPM may be a promising imaging technique for detecting tumor cells in esophageal cancer.

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

Science.gov (United States)

Chen, Sijia; Yang, Yinghong; Jiang, Weizhong; Feng, Changyin; Chen, Zhifen; Zhuo, Shuangmu; Zhu, Xiaoqin; Guan, Guoxian; Chen, Jianxin

2014-10-01

The examination of stromal fibrosis within colorectal cancer is overlooked, not only because the routine pathological examinations seem to focus more on tumour staging and precise surgical margins, but also because of the lack of efficient diagnostic methods. Multiphoton microscopy (MPM) can be used to study the muscularis stroma of normal and colorectal carcinoma tissue at the molecular level. In this work, we attempt to show the feasibility of MPM for discerning the microstructure of the normal human rectal muscle layer and fibrosis colorectal carcinoma tissue practicably. Three types of muscularis propria stromal fibrosis beneath the colorectal cancer infiltration were first observed through the MPM imaging system by providing intercellular microstructural details in fresh, unstained tissue samples. Our approach also presents the capability of quantifying the extent of stromal fibrosis from both amount and orientation of collagen, which may further characterize the severity of fibrosis. By comparing with the pathology analysis, these results show that the MPM has potential advantages in becoming a histological tool for detecting the stromal fibrosis and collecting prognosis evidence, which may guide subsequent therapy procedures for patients into good prognosis.

Real-time visualization of melanin granules in normal human skin using combined multiphoton and reflectance confocal microscopy.

Science.gov (United States)

Majdzadeh, Ali; Lee, Anthony M D; Wang, Hequn; Lui, Harvey; McLean, David I; Crawford, Richard I; Zloty, David; Zeng, Haishan

2015-05-01

Recent advances in biomedical optics have enabled dermal and epidermal components to be visualized at subcellular resolution and assessed noninvasively. Multiphoton microscopy (MPM) and reflectance confocal microscopy (RCM) are noninvasive imaging modalities that have demonstrated promising results in imaging skin micromorphology, and which provide complementary information regarding skin components. This study assesses whether combined MPM/RCM can visualize intracellular and extracellular melanin granules in the epidermis and dermis of normal human skin. We perform MPM and RCM imaging of in vivo and ex vivo skin in the infrared domain. The inherent three-dimensional optical sectioning capability of MPM/RCM is used to image high-contrast granular features across skin depths ranging from 50 to 90 μm. The optical images thus obtained were correlated with conventional histologic examination including melanin-specific staining of ex vivo specimens. MPM revealed highly fluorescent granular structures below the dermal-epidermal junction (DEJ) region. Histochemical staining also demonstrated melanin-containing granules that correlate well in size and location with the granular fluorescent structures observed in MPM. Furthermore, the MPM fluorescence excitation wavelength and RCM reflectance of cell culture-derived melanin were equivalent to those of the granules. This study suggests that MPM can noninvasively visualize and quantify subepidermal melanin in situ. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

A national facility for biological cryo-electron microscopy

International Nuclear Information System (INIS)

Saibil, Helen R.; Grünewald, Kay; Stuart, David I.

2015-01-01

This review provides a brief update on the use of cryo-electron microscopy for integrated structural biology, along with an overview of the plans for the UK national facility for electron microscopy being built at the Diamond synchrotron. Three-dimensional electron microscopy is an enormously powerful tool for structural biologists. It is now able to provide an understanding of the molecular machinery of cells, disease processes and the actions of pathogenic organisms from atomic detail through to the cellular context. However, cutting-edge research in this field requires very substantial resources for equipment, infrastructure and expertise. Here, a brief overview is provided of the plans for a UK national three-dimensional electron-microscopy facility for integrated structural biology to enable internationally leading research on the machinery of life. State-of-the-art equipment operated with expert support will be provided, optimized for both atomic-level single-particle analysis of purified macromolecules and complexes and for tomography of cell sections. The access to and organization of the facility will be modelled on the highly successful macromolecular crystallography (MX) synchrotron beamlines, and will be embedded at the Diamond Light Source, facilitating the development of user-friendly workflows providing near-real-time experimental feedback

Multimodal imaging of vocal fold scarring in a rabbit model by multiphoton microscopy

Science.gov (United States)

Kazarine, Alexei; Bouhabel, Sarah; Douillette, Annie H.; Kost, Karen; Li-Jessen, Nicole Y. K.; Mongeau, Luc; Wiseman, Paul W.

2017-02-01

Vocal fold scarring as a result of injury or disease can lead to voice disorders which can significantly affect the quality of life. During the scarring process, the normally elastic tissue of the vocal fold lamina propria is replaced by a much stiffer collagen-based fibrotic tissue, which impacts the fold's ability to vibrate. Surgical removal of this tissue is often ineffective and can result in further scarring. Injectable biomaterials, a form of tissue engineering, have been proposed as a potential solution to reduce existing scars or prevent scarring altogether. In order to properly evaluate the effectiveness of these new materials, multiphoton microscopy emerges as an effective tool due to its intrinsic multiple label free contrast mechanisms that highlight extracellular matrix elements. In this study, we evaluate the spatial distribution of collagen and elastin fibers in a rabbit model using second harmonic generation (SHG), third harmonic generation (THG) and two photon autofluorescence (TPAF) applied to unlabeled tissue sections. In comparison to traditional methods that rely on histological staining or immunohistochemistry, SHG, THG and TPAF provide a more reliable detection of these native proteins. The evaluation of collagen levels allows us to follow the extent of scarring, while the presence of elastin fibers is thought to be indicative of the level of healing of the injured fold. Using these imaging modalities, we characterize the outcome of injectable biomaterial treatments in order to direct future treatments for tissue engineering.

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

Science.gov (United States)

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

2016-03-01

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

Multi-photon microscope driven by novel green laser pump

Science.gov (United States)

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

2016-03-01

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

Optimisation-based wavefront sensorless adaptive optics for microscopy

NARCIS (Netherlands)

Antonello, J.

2014-01-01

Microscopy is an essential tool for life sciences. Thanks to the development of confocal and multiphoton microscopy, scientists are able to obtain high-resolution 3D views of biological specimens. Nevertheless, spatial variations in the index of refraction within specimens cause aberrations that

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

DEFF Research Database (Denmark)

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

2011-01-01

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

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

Science.gov (United States)

Chen, Youting; Chen, Jing; Chen, Hong; Hong, Zhipeng; Zhu, Xiaoqin; Zhuo, Shuangmu; Chen, Yanling; Chen, Jianxin

2014-09-01

Hematoxylin and eosin (H&E) staining of tissue samples is the standard approach in histopathology for imaging and diagnosing cancer. Recent reports have shown that multiphoton microscopy (MPM) provides better sample interface with single-cell resolution, which enhances traditional H&E staining and offers a powerful diagnostic tool with potential applications in oncology. The purpose of this study was to further expand the versatility of MPM by establishing the optical parameters required for imaging unstained histological sections of pancreatic neoplasms, thereby providing an efficient and environmentally sustainable alternative to H&E staining while improving the accuracy of pancreatic cancer diagnoses. We found that the high-resolution MPM images clearly distinguish between the structure of normal pancreatic tissues compared with pancreatic neoplasms in unstained histological sections, and discernable differences in tissue architecture and cell morphology between normal versus tumorigenic cells led to enhanced optical diagnosis of cancerous tissue. Moreover, quantitative assessment of the cytomorphological features visualized from MPM images showed significant differences in the nuclear-cytoplasmic ratios of pancreatic neoplasms compared with normal pancreas, as well as further distinguished pancreatic malignant tumors from benign tumors. These results indicate that the MPM could potentially serve as an optical tool for the diagnosis of pancreatic neoplasms in unstained histological sections.

Multiphoton quantum optics and quantum state engineering

International Nuclear Information System (INIS)

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

2006-01-01

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

Multiphoton quantum optics and quantum state engineering

Energy Technology Data Exchange (ETDEWEB)

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

2006-05-15

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

Multiphoton fluorescence lifetime imaging of chemotherapy distribution in solid tumors

Science.gov (United States)

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

2017-11-01

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

Identification of the boundary between normal breast tissue and invasive ductal carcinoma during breast-conserving surgery using multiphoton microscopy

Science.gov (United States)

Deng, Tongxin; Nie, Yuting; Lian, Yuane; Wu, Yan; Fu, Fangmeng; Wang, Chuan; Zhuo, Shuangmu; Chen, Jianxin

2014-11-01

Breast-conserving surgery has become an important way of surgical treatment for breast cancer worldwide nowadays. Multiphoton microscopy (MPM) has the ability to noninvasively visualize tissue architectures at the cellular level using intrinsic fluorescent molecules in biological tissues without the need for fluorescent dye. In this study, MPM is used to image the microstructures of terminal duct lobular unit (TDLU), invasive ductal carcinoma and the boundary region between normal and cancerous breast tissues. Our study demonstrates that MPM has the ability to not only reveal the morphological changes of the cuboidal epithelium, basement membrane and interlobular stroma but also identify the boundary between normal breast tissue and invasive ductal carcinoma, which correspond well to the Hematoxylin and Eosin (H and E) images. Predictably, MPM can monitor surgical margins in real time and provide considerable accuracy for resection of breast cancerous tissues intraoperatively. With the development of miniature, real-time MPM imaging technology, MPM should have great application prospects during breast-conserving surgery.

Multi-Photon Entanglement and Quantum Teleportation

National Research Council Canada - National Science Library

Shih, Yanhua

1999-01-01

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

Super-resolution fluorescence microscopy by stepwise optical saturation

Science.gov (United States)

Zhang, Yide; Nallathamby, Prakash D.; Vigil, Genevieve D.; Khan, Aamir A.; Mason, Devon E.; Boerckel, Joel D.; Roeder, Ryan K.; Howard, Scott S.

2018-01-01

Super-resolution fluorescence microscopy is an important tool in biomedical research for its ability to discern features smaller than the diffraction limit. However, due to its difficult implementation and high cost, the super-resolution microscopy is not feasible in many applications. In this paper, we propose and demonstrate a saturation-based super-resolution fluorescence microscopy technique that can be easily implemented and requires neither additional hardware nor complex post-processing. The method is based on the principle of stepwise optical saturation (SOS), where M steps of raw fluorescence images are linearly combined to generate an image with a M-fold increase in resolution compared with conventional diffraction-limited images. For example, linearly combining (scaling and subtracting) two images obtained at regular powers extends the resolution by a factor of 1.4 beyond the diffraction limit. The resolution improvement in SOS microscopy is theoretically infinite but practically is limited by the signal-to-noise ratio. We perform simulations and experimentally demonstrate super-resolution microscopy with both one-photon (confocal) and multiphoton excitation fluorescence. We show that with the multiphoton modality, the SOS microscopy can provide super-resolution imaging deep in scattering samples. PMID:29675306

Networking strategies of the microscopy community for improved utilization of advanced instruments: (1) The Australian Microscopy and Microanalysis Research Facility (AMMRF)

International Nuclear Information System (INIS)

Ringer, S.P.; Apperley, M.H.

2014-01-01

This paper describes the strategy underpinning the formation and operation of the Australian Microscopy and Microanalysis Research Facility (AMMRF). AMMRF is a formal collaboration that links eight Australian Universities together to create a user-focused national capability in microscopy and microanalysis. The AMMRF flagship capabilities include: Cameca IMS-1280 and NanoSIMS-50 ion microprobes (University of Western Australia); High-throughput, high-resolution cryoTEM (University of Queensland); Atom Probe Microscopy (University of Sydney); High-resolution Focussed Ion-Beam and SEM (Universities of Adelaide and NSW); High-resolution SEM microanalysis facility (University of New South Wales); and PHI TRIFT V nanoToF ToF-SIMS (University of South Australia). Secondly, a network of peer support and expert training has been established amongst facility professional support staff. The governance and funding of the organisation are described and the advantages and achievements of a nationally coordinated facility for microscopy and microanalysis are set out. Selected data are presented that benchmark the performance of the facility, describe the economic impact and demonstrate the impact on the quality of research outcomes as a result of operating national collaborative research infrastructure for microscopy and microanalysis

High-resolution imaging of basal cell carcinoma: a comparison between multiphoton microscopy with fluorescence lifetime imaging and reflectance confocal microscopy.

Science.gov (United States)

Manfredini, Marco; Arginelli, Federica; Dunsby, Christopher; French, Paul; Talbot, Clifford; König, Karsten; Pellacani, Giovanni; Ponti, Giovanni; Seidenari, Stefania

2013-02-01

The aim of this study was to compare morphological aspects of basal cell carcinoma (BCC) as assessed by two different imaging methods: in vivo reflectance confocal microscopy (RCM) and multiphoton tomography with fluorescence lifetime imaging implementation (MPT-FLIM). The study comprised 16 BCCs for which a complete set of RCM and MPT-FLIM images were available. The presence of seven MPT-FLIM descriptors was evaluated. The presence of seven RCM equivalent parameters was scored in accordance to their extension. Chi-squared test with Fisher's exact test and Spearman's rank correlation coefficient were determined between MPT-FLIM scores and adjusted-RCM scores. MPT-FLIM and RCM descriptors of BCC were coupled to match the descriptors that define the same pathological structures. The comparison included: Streaming and Aligned elongated cells, Streaming with multiple directions and Double alignment, Palisading (RCM) and Palisading (MPT-FLIM), Typical tumor islands, and Cell islands surrounded by fibers, Dark silhouettes and Phantom islands, Plump bright cells and Melanophages, Vessels (RCM), and Vessels (MPT-FLIM). The parameters that were significantly correlated were Melanophages/Plump Bright Cells, Aligned elongated cells/Streaming, Double alignment/Streaming with multiple directions, and Palisading (MPT-FLIM)/Palisading (RCM). According to our data, both methods are suitable to image BCC's features. The concordance between MPT-FLIM and RCM is high, with some limitations due to the technical differences between the two devices. The hardest difficulty when comparing the images generated by the two imaging modalities is represented by their different field of view. © 2012 John Wiley & Sons A/S.

Quantification of aortic and cutaneous elastin and collagen morphology in Marfan syndrome by multiphoton microscopy.

Science.gov (United States)

Cui, Jason Z; Tehrani, Arash Y; Jett, Kimberly A; Bernatchez, Pascal; van Breemen, Cornelis; Esfandiarei, Mitra

2014-09-01

In a mouse model of Marfan syndrome, conventional Verhoeff-Van Gieson staining displays severe fragmentation, disorganization and loss of the aortic elastic fiber integrity. However, this method involves chemical fixatives and staining, which may alter the native morphology of elastin and collagen. Thus far, quantitative analysis of fiber damage in aorta and skin in Marfan syndrome has not yet been explored. In this study, we have used an advanced noninvasive and label-free imaging technique, multiphoton microscopy to quantify fiber fragmentation, disorganization, and total volumetric density of aortic and cutaneous elastin and collagen in a mouse model of Marfan syndrome. Aorta and skin samples were harvested from Marfan and control mice aged 3-, 6- and 9-month. Elastin and collagen were identified based on two-photon excitation fluorescence and second-harmonic-generation signals, respectively, without exogenous label. Measurement of fiber length indicated significant fragmentation in Marfan vs. control. Fast Fourier transform algorithm analysis demonstrated markedly lower fiber organization in Marfan mice. Significantly reduced volumetric density of elastin and collagen and thinner skin dermis were observed in Marfan mice. Cutaneous content of elastic fibers and thickness of dermis in 3-month Marfan resembled those in the oldest control mice. Our findings of early signs of fiber degradation and thinning of skin dermis support the potential development of a novel non-invasive approach for early diagnosis of Marfan syndrome. Copyright © 2014 Elsevier Inc. All rights reserved.

Resolution improvement by nonconfocal theta microscopy.

Science.gov (United States)

Lindek, S; Stelzer, E H

1999-11-01

We present a novel scanning fluorescence microscopy technique, nonconfocal theta microscopy (NCTM), that provides almost isotropic resolution. In NCTM, multiphoton absorption from two orthogonal illumination directions is used to induce fluorescence emission. Therefore the point-spread function of the microscope is described by the product of illumination point-spread functions with reduced spatial overlap, which provides the resolution improvement and the more isotropic observation volume. We discuss the technical details of this new method.

Laser-induced multiphoton transitions

International Nuclear Information System (INIS)

Stenholm, S.

1978-06-01

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

Multiphoton states and amplitude k-th power squeezing

International Nuclear Information System (INIS)

Buzek, V.; Jex, I.

1991-01-01

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

Clinical multiphoton FLIM tomography

Science.gov (United States)

König, Karsten

2012-03-01

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

Multiphoton Processes and Attosecond Physics

CERN Document Server

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

2012-01-01

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

A national facility for biological cryo-electron microscopy.

Science.gov (United States)

Saibil, Helen R; Grünewald, Kay; Stuart, David I

2015-01-01

Three-dimensional electron microscopy is an enormously powerful tool for structural biologists. It is now able to provide an understanding of the molecular machinery of cells, disease processes and the actions of pathogenic organisms from atomic detail through to the cellular context. However, cutting-edge research in this field requires very substantial resources for equipment, infrastructure and expertise. Here, a brief overview is provided of the plans for a UK national three-dimensional electron-microscopy facility for integrated structural biology to enable internationally leading research on the machinery of life. State-of-the-art equipment operated with expert support will be provided, optimized for both atomic-level single-particle analysis of purified macromolecules and complexes and for tomography of cell sections. The access to and organization of the facility will be modelled on the highly successful macromolecular crystallography (MX) synchrotron beamlines, and will be embedded at the Diamond Light Source, facilitating the development of user-friendly workflows providing near-real-time experimental feedback.

New developments in multimodal clinical multiphoton tomography

Science.gov (United States)

König, Karsten

2011-03-01

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

Multiphoton bibliography

International Nuclear Information System (INIS)

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

1981-12-01

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

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

Science.gov (United States)

Bancelin, Stéphane; Lynch, Barbara; Bonod-Bidaud, Christelle; Ducourthial, Guillaume; Psilodimitrakopoulos, Sotiris; Dokládal, Petr; Allain, Jean-Marc; Schanne-Klein, Marie-Claire; Ruggiero, Florence

2015-12-01

Soft connective tissues such as skin, tendon or cornea are made of about 90% of extracellular matrix proteins, fibrillar collagens being the major components. Decreased or aberrant collagen synthesis generally results in defective tissue mechanical properties as the classic form of Elhers-Danlos syndrome (cEDS). This connective tissue disorder is caused by mutations in collagen V genes and is mainly characterized by skin hyperextensibility. To investigate the relationship between the microstructure of normal and diseased skins and their macroscopic mechanical properties, we imaged and quantified the microstructure of dermis of ex vivo murine skin biopsies during uniaxial mechanical assay using multiphoton microscopy. We used two genetically-modified mouse lines for collagen V: a mouse model for cEDS harboring a Col5a2 deletion (a.k.a. pN allele) and the transgenic K14-COL5A1 mice which overexpress the human COL5A1 gene in skin. We showed that in normal skin, the collagen fibers continuously align with stretch, generating the observed increase in mechanical stress. Moreover, dermis from both transgenic lines exhibited altered collagen reorganization upon traction, which could be linked to microstructural modifications. These findings show that our multiscale approach provides new crucial information on the biomechanics of dermis that can be extended to all collagen-rich soft tissues.

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

Energy Technology Data Exchange (ETDEWEB)

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

2011-07-01

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

Use of multi-photon laser-scanning microscopy to describe the distribution of xenobiotic chemicals in fish early life stages

International Nuclear Information System (INIS)

Hornung, Michael W.; Cook, Philip M.; Flynn, Kevin M.; Lothenbach, Doug B.; Johnson, Rodney D.; Nichols, John W.

2004-01-01

To better understand the mechanisms by which persistent bioaccumulative toxicants (PBTs) produce toxicity during fish early life stages (ELS), dose-response relationships need to be understood in relation to the dynamic distribution of chemicals in sensitive tissues. In this study, a multi-photon laser scanning microscope (MPLSM) was used to determine the multi-photon excitation spectra of several polyaromatic hydrocarbons (PAHs) and to describe chemical distribution among tissues during fish ELS. The multi-photon excitation spectra revealed intense fluorescent signal from the model fluorophore, pentamethyl-difluoro-boro-indacene (BODIPY[reg], less signal from benzo[a]pyrene and fluoranthene, and no detectable signal from pyrene. The imaging method was tested by exposing newly fertilized medaka (Oryzias latipes) eggs to BODIPY[reg] or fluoranthene for 6 h, followed by transfer to clean media. Embryos and larvae were then imaged through 5 days post-hatch. The two test chemicals partitioned similarly throughout development and differences in fluorescence intensity among tissues were evident to a depth of several hundred microns. Initially, the most intense signal was observed in the oil droplet within the yolk, while a moderate signal was seen in the portion of the yolk containing the yolk-platelets. As embryonic development progressed, the liver biliary system, gall bladder, and intestinal tract accumulated strong fluorescent signal. After hatch, once the gastrointestinal tract was completely developed, most of the fluorescent signal was cleared. The MPLSM is a useful tool to describe the tissue distribution of fluorescent PBTs during fish ELS

High-resolution multimodal clinical multiphoton tomography of skin

Science.gov (United States)

König, Karsten

2011-03-01

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

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

Science.gov (United States)

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

2010-02-01

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

Setting up and running an advanced light microscopy and imaging facility.

Science.gov (United States)

Sánchez, Carlos; Muñoz, Ma Ángeles; Villalba, Maite; Labrador, Verónica; Díez-Guerra, F Javier

2011-07-01

During the last twenty years, interest in light microscopy and imaging techniques has grown in various fields, such as molecular and cellular biology, developmental biology, and neurobiology. In addition, the number of scientific articles and journals using these techniques is rapidly increasing. Nowadays, most research institutions require sophisticated microscopy systems to cover their investigation demands. In general, such instruments are too expensive and complex to be purchased and managed by a single laboratory or research group, so they have to be shared with other groups and supervised by specialized personnel. This is the reason why microscopy and imaging facilities are becoming so important at research institutions nowadays. In this unit, we have gathered and presented a number of issues and considerations from our own experience that we hope will be helpful when planning or setting up a new facility.

In vivo multiphoton-microscopy of picosecond-laser-induced optical breakdown in human skin.

Science.gov (United States)

Balu, Mihaela; Lentsch, Griffin; Korta, Dorota Z; König, Karsten; Kelly, Kristen M; Tromberg, Bruce J; Zachary, Christopher B

2017-08-01

Improvements in skin appearance resulting from treatment with fractionated picosecond-lasers have been noted, but optimizing the treatment efficacy depends on a thorough understanding of the specific skin response. The development of non-invasive laser imaging techniques in conjunction with laser therapy can potentially provide feedback for guidance and optimizing clinical outcome. The purpose of this study was to demonstrate the capability of multiphoton microscopy (MPM), a high-resolution, label-free imaging technique, to characterize in vivo the skin response to a fractionated non-ablative picosecond-laser treatment. Two areas on the arm of a volunteer were treated with a fractionated picosecond laser at the Dermatology Clinic, UC Irvine. The skin response to treatment was imaged in vivo with a clinical MPM-based tomograph at 3 hours and 24 hours after treatment and seven additional time points over a 4-week period. MPM revealed micro-injuries present in the epidermis. Pigmented cells were particularly damaged in the process, suggesting that melanin is likely the main absorber for laser induced optical breakdown. Damaged individual cells were distinguished as early as 3 hours post pico-laser treatment with the 532 nm wavelength, and 24 hours post-treatment with both 532 and 1064 nm wavelengths. At later time points, clusters of cellular necrotic debris were imaged across the treated epidermis. After 24 hours of treatment, inflammatory cells were imaged in the proximity of epidermal micro-injuries. The epidermal injuries were exfoliated over a 4-week period. This observational and descriptive pilot study demonstrates that in vivo MPM imaging can be used non-invasively to provide label-free contrast for describing changes in human skin following a fractionated non-ablative laser treatment. The results presented in this study represent the groundwork for future longitudinal investigations on an expanded number of subjects to understand the response to treatment

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

Science.gov (United States)

McMullen, Jesse D; Zipfel, Warren R

2010-03-15

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

Concepts in Light Microscopy of Viruses

Science.gov (United States)

Witte, Robert; Georgi, Fanny

2018-01-01

Viruses threaten humans, livestock, and plants, and are difficult to combat. Imaging of viruses by light microscopy is key to uncover the nature of known and emerging viruses in the quest for finding new ways to treat viral disease and deepening the understanding of virus–host interactions. Here, we provide an overview of recent technology for imaging cells and viruses by light microscopy, in particular fluorescence microscopy in static and live-cell modes. The review lays out guidelines for how novel fluorescent chemical probes and proteins can be used in light microscopy to illuminate cells, and how they can be used to study virus infections. We discuss advantages and opportunities of confocal and multi-photon microscopy, selective plane illumination microscopy, and super-resolution microscopy. We emphasize the prevalent concepts in image processing and data analyses, and provide an outlook into label-free digital holographic microscopy for virus research. PMID:29670029

Amplitudes for multiphoton quantum processes in linear optics

International Nuclear Information System (INIS)

UrIas, Jesus

2011-01-01

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

Amplitudes for multiphoton quantum processes in linear optics

Science.gov (United States)

Urías, Jesús

2011-07-01

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

Delivery of cyclodextrin polymers to bacterial biofilms - An exploratory study using rhodamine labelled cyclodextrins and multiphoton microscopy.

Science.gov (United States)

Thomsen, Hanna; Benkovics, Gábor; Fenyvesi, Éva; Farewell, Anne; Malanga, Milo; Ericson, Marica B

2017-10-15

Cyclodextrin (CD) polymers are interesting nanoparticulate systems for pharmaceutical delivery; however, knowledge regarding their applications towards delivery into complex microbial biofilm structures is so far limited. The challenge is to demonstrate penetration and transport through the biofilm and its exopolysaccharide matrix. The ideal functionalization for penetration into mature biofilms is unexplored. In this paper, we present a novel set of rhodamine labelled βCD-polymers, with different charge moieties, i.e., neutral, anionic, and cationic, and explore their potential delivery into mature Staphylococcus epidermidis biofilms using multiphoton laser scanning microscopy (MPM). The S. epidermidis biofilms, being a medically relevant model organism, were stained with SYTO9. By using MPM, three-dimensional imaging and spectral investigation of the distribution of the βCD-polymers could be obtained. It was found that the cationic βCD-polymers showed significantly higher integration into the biofilms, compared to neutral and anionic functionalized βCDs. None of the carriers presented any inherent toxicity to the biofilms, meaning that the addition of rhodamine moiety does not affect the inertness of the delivery system. Taken together, this study demonstrates a novel approach by which delivery of fluorescently labelled CD nanoparticles to bacterial biofilms can be explored using MPM. Future studies should be undertaken investigating the potential in using cationic functionalization of CD based delivery systems for targeting anti-microbial effects in biofilms. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

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

Science.gov (United States)

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

2016-01-01

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

Evaluation of multiphoton effects in down-conversion

International Nuclear Information System (INIS)

Yoshimi, Kazuyoshi; Koshino, Kazuki

2010-01-01

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

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

Directory of Open Access Journals (Sweden)

Michael G Giacomelli

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

The X-Ray Microscopy And Micro-Spectroscopy Facility At The ESRF

International Nuclear Information System (INIS)

Susini, J.; Somogyi, A.; Barrett, R.; Salome, M.; Bohic, S.; Fayard, B.; Eichert, D.; Dhez, O.; Bleuet, P.; Martinez-Criado, G.; Tucoulou, R.

2004-01-01

Among the 40 beamlines in operation at the European Synchrotron Radiation Facility, three beamlines are fully dedicated to X-ray microscopy and micro-spectroscopy techniques in the multi-keV range. Offering a unique combination of non destructive analytical techniques which aim to satisfy the growing demand from experimental research fields such as medicine, geology, archaeology, earth, planetary and environmental sciences. Following a brief discussion on the strengths and weaknesses of X-ray microscopy and spectro-microscopy techniques in the 1-20keV range, characteristics of the beamlines are briefly described. Examples of applications are given in the reference list

Multiphoton resonances

International Nuclear Information System (INIS)

Shore, B.W.

1977-01-01

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

Multi-photon excitation microscopy for advanced biomedical imaging

NARCIS (Netherlands)

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

Fluorescence microscopy (FM) is a technique traditionally used for determining biological structures [33]; its basic concept is summarised in Figure 1a. The biological specimen under examination is labelled with one or more fluorescent probes before being placed in the microscope. A single photon

Super-nonlinear fluorescence microscopy for high-contrast deep tissue imaging

Science.gov (United States)

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

2014-02-01

Two-photon excited fluorescence microscopy (TPFM) offers the highest penetration depth with subcellular resolution in light microscopy, due to its unique advantage of nonlinear excitation. However, a fundamental imaging-depth limit, accompanied by a vanishing signal-to-background contrast, still exists for TPFM when imaging deep into scattering samples. Formally, the focusing depth, at which the in-focus signal and the out-of-focus background are equal to each other, is defined as the fundamental imaging-depth limit. To go beyond this imaging-depth limit of TPFM, we report a new class of super-nonlinear fluorescence microscopy for high-contrast deep tissue imaging, including multiphoton activation and imaging (MPAI) harnessing novel photo-activatable fluorophores, stimulated emission reduced fluorescence (SERF) microscopy by adding a weak laser beam for stimulated emission, and two-photon induced focal saturation imaging with preferential depletion of ground-state fluorophores at focus. The resulting image contrasts all exhibit a higher-order (third- or fourth- order) nonlinear signal dependence on laser intensity than that in the standard TPFM. Both the physical principles and the imaging demonstrations will be provided for each super-nonlinear microscopy. In all these techniques, the created super-nonlinearity significantly enhances the imaging contrast and concurrently extends the imaging depth-limit of TPFM. Conceptually different from conventional multiphoton processes mediated by virtual states, our strategy constitutes a new class of fluorescence microscopy where high-order nonlinearity is mediated by real population transfer.

QED theory of multiphoton transitions in atoms and ions

Science.gov (United States)

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

2018-03-01

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

Multiphoton microscopy observations of 3D elastin and collagen fiber microstructure changes during pressurization in aortic media.

Science.gov (United States)

Sugita, Shukei; Matsumoto, Takeo

2017-06-01

Elastin and collagen fibers play important roles in the mechanical properties of aortic media. Because knowledge of local fiber structures is required for detailed analysis of blood vessel wall mechanics, we investigated 3D microstructures of elastin and collagen fibers in thoracic aortas and monitored changes during pressurization. Using multiphoton microscopy, autofluorescence images from elastin and second harmonic generation signals from collagen were acquired in media from rabbit thoracic aortas that were stretched biaxially to restore physiological dimensions. Both elastin and collagen fibers were observed in all longitudinal-circumferential plane images, whereas alternate bright and dark layers were observed along the radial direction and were recognized as elastic laminas (ELs) and smooth muscle-rich layers (SMLs), respectively. Elastin and collagen fibers are mainly oriented in the circumferential direction, and waviness of collagen fibers was significantly higher than that of elastin fibers. Collagen fibers were more undulated in longitudinal than in radial direction, whereas undulation of elastin fibers was equibiaxial. Changes in waviness of collagen fibers during pressurization were then evaluated using 2-dimensional fast Fourier transform in mouse aortas, and indices of waviness of collagen fibers decreased with increases in intraluminal pressure. These indices also showed that collagen fibers in SMLs became straight at lower intraluminal pressures than those in EL, indicating that SMLs stretched more than ELs. These results indicate that deformation of the aorta due to pressurization is complicated because of the heterogeneity of tissue layers and differences in elastic properties of ELs, SMLs, and surrounding collagen and elastin.

In vivo multiphoton imaging of bile duct ligation

Science.gov (United States)

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

2008-02-01

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

Design and development of compact multiphoton microscopes

Science.gov (United States)

Mehravar, SeyedSoroush

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

Multiphoton excitation and high-harmonics generation in topological insulator.

Science.gov (United States)

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

2018-05-10

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

Multiphoton excitation and high-harmonics generation in topological insulator

Science.gov (United States)

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

2018-05-01

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

Spin Multiphoton Antiresonance at Finite Temperatures

Science.gov (United States)

Hicke, Christian; Dykman, Mark

2007-03-01

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

Multiphotonic resonance processes in potassium vapor

International Nuclear Information System (INIS)

Bensoussan, Paul.

1975-01-01

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

Comparison of higher-order multiphoton signal generation and collection at the 1700-nm window based on transmittance measurement of objective lenses.

Science.gov (United States)

Wen, Wenhui; Wang, Yuxin; Liu, Hongji; Wang, Kai; Qiu, Ping; Wang, Ke

2018-01-01

One benefit of excitation at the 1700-nm window is the more accessible modalities of multiphoton signal generation. It is demonstrated here that the transmittance performance of the objective lens is of vital importance for efficient higher-order multiphoton signal generation and collection excited at the 1700-nm window. Two commonly used objective lenses for multiphoton microscopy (MPM) are characterized and compared, one with regular coating and the other with customized coating for high transmittance at the 1700-nm window. Our results show that, fourth harmonic generation imaging of mouse tail tendon and 5-photon fluorescence of carbon quantum dots using the regular objective lens shows an order of magnitude signal higher than those using the customized objective lens. Besides, the regular objective lens also enables a 3-photon fluorescence imaging depth of >1600 μm in mouse brain in vivo. Our results will provide guidelines for objective lens selection for MPM at the 1700-nm window. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

External quality assessment of malaria microscopy diagnosis in selected health facilities in Western Oromia, Ethiopia.

Science.gov (United States)

Sori, Getachew; Zewdie, Olifan; Tadele, Geletta; Samuel, Abdi

2018-06-18

Accurate early diagnosis and prompt treatment are one of the key strategies to control and prevent malaria disease. External quality assessment is the most effective method for evaluation of the quality of malaria microscopy diagnosis. The aim of this study was to assess the quality of malaria microscopy diagnosis and its associated factors in selected public health facility laboratories in East Wollega Zone, Western Ethiopia. Facility-based cross-sectional study design was conducted in 30 randomly selected public health facility laboratories from November 2014 to January 2015 in East Wollega Zone, Western Ethiopia. Ten validated stained malaria panel slides with known Plasmodium species, developmental stage and parasite density were distributed. Data were captured; cleaned and analyzed using SPSS version 20 statistical software-multivariate logistic regressions and the agreement in reading between the peripheral diagnostic centers and the reference laboratory were done using kappa statistics. A total of 30 health facility laboratories were involved in the study and the overall quality of malaria microscopy diagnosis was poor (62.3%). The associated predictors of quality in this diagnosis were in-service training [(AOR = 16, 95% CI (1.3, 1.96)], smearing quality [(AOR = 24, 95% CI (1.8, 3.13)], staining quality [(AOR = 15, 95% CI (2.35, 8.61), parasite detection [(AOR = 9, 95% CI (1.1, 8.52)] and identification skills [(AOR = 8.6, 95% CI (1.21, 1.63)]. Eighteen (60%) of health facility laboratories had in-service trained laboratory professionals on malaria microscopy diagnosis. Overall quality of malaria microscopy diagnosis was poor and a significant gap in this service was observed that could impact on its diagnostic services.

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

International Nuclear Information System (INIS)

Wang, Yuxin; Wen, Wenhui; Wang, Kai; Wang, Ke; Zhai, Peng; Qiu, Ping

2016-01-01

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

Scanning transmission ion microscopy on Fudan SPM facility

International Nuclear Information System (INIS)

Li Yongqiang; Shen Hao; Zheng Yi; Li Xinyi; Liu Bo; Satoh Takahiro

2011-01-01

In this paper, we report a novel measurement system based on the development of Fudan Scanning Proton Microscopy (SPM) facility. By using Si-PIN diode(Hamamatsu S1223-01) detector, scanning transmission ion microscopy (STIM) measurement system has been set up. It can provide density and structural images with high probing efficiency and non-destruction by utilizing the energy loss of high energy (MeV) and focused ions penetrating through a thin sample. STIM measurement is able to map the density distribution of organic elements which mostly compose biology materials, such information can not be detected by using conventional Be-windowed Si (Li) X-ray detector in Particle Induced X-ray Emission (PIXE) technique. The spatial resolution capability of STIM is higher than PIXE technique at same accelerator status. As a result of STIM measurement, Paramecium attached on the top of Kapton tube was measured by STIM. (authors)

Modulation of the pupil function of microscope objective lens for multifocal multi-photon microscopy using a spatial light modulator

Science.gov (United States)

Matsumoto, Naoya; Okazaki, Shigetoshi; Takamoto, Hisayoshi; Inoue, Takashi; Terakawa, Susumu

2014-02-01

We propose a method for high precision modulation of the pupil function of a microscope objective lens to improve the performance of multifocal multi-photon microscopy (MMM). To modulate the pupil function, we adopt a spatial light modulator (SLM) and place it at the conjugate position of the objective lens. The SLM can generate an arbitrary number of spots to excite the multiple fluorescence spots (MFS) at the desired positions and intensities by applying an appropriate computer-generated hologram (CGH). This flexibility allows us to control the MFS according to the photobleaching level of a fluorescent protein and phototoxicity of a specimen. However, when a large number of excitation spots are generated, the intensity distribution of the MFS is significantly different from the one originally designed due to misalignment of the optical setup and characteristics of the SLM. As a result, the image of a specimen obtained using laser scanning for the MFS has block noise segments because the SLM could not generate a uniform MFS. To improve the intensity distribution of the MFS, we adaptively redesigned the CGH based on the observed MFS. We experimentally demonstrate an improvement in the uniformity of a 10 × 10 MFS grid using a dye solution. The simplicity of the proposed method will allow it to be applied for calibration of MMM before observing living tissue. After the MMM calibration, we performed laser scanning with two-photon excitation to observe a real specimen without detecting block noise segments.

Limiting energy loss distributions for multiphoton channeling radiation

International Nuclear Information System (INIS)

Bondarenco, M.V.

2015-01-01

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

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

Science.gov (United States)

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

2016-03-01

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

The importance of spectroscopy for infrared multiphoton excitation

International Nuclear Information System (INIS)

Fuss, W.; Kompa, K.L.

1980-07-01

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

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

Science.gov (United States)

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

2016-05-01

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

Transverse correlations in multiphoton entanglement

International Nuclear Information System (INIS)

Wen Jianming; Rubin, Morton H.; Shih Yanhua

2007-01-01

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

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

Energy Technology Data Exchange (ETDEWEB)

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

2009-12-15

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

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

Energy Technology Data Exchange (ETDEWEB)

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

2011-01-17

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

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

International Nuclear Information System (INIS)

Idrish Miah, M.

2011-01-01

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

Temporal focusing-based widefield multiphoton microscopy with spatially modulated illumination for biotissue imaging.

Science.gov (United States)

Chang, Chia-Yuan; Lin, Cheng-Han; Lin, Chun-Yu; Sie, Yong-Da; Hu, Yvonne Yuling; Tsai, Sheng-Feng; Chen, Shean-Jen

2018-01-01

A developed temporal focusing-based multiphoton excitation microscope (TFMPEM) has a digital micromirror device (DMD) which is adopted not only as a blazed grating for light spatial dispersion but also for patterned illumination simultaneously. Herein, the TFMPEM has been extended to implement spatially modulated illumination at structured frequency and orientation to increase the beam coverage at the back-focal aperture of the objective lens. The axial excitation confinement (AEC) of TFMPEM can be condensed from 3.0 μm to 1.5 μm for a 50 % improvement. By using the TFMPEM with HiLo technique as two structured illuminations at the same spatial frequency but different orientation, reconstructed biotissue images according to the condensed AEC structured illumination are shown obviously superior in contrast and better scattering suppression. Picture: TPEF images of the eosin-stained mouse cerebellar cortex by conventional TFMPEM (left), and the TFMPEM with HiLo technique as 1.09 μm -1 spatially modulated illumination at 90° (center) and 0° (right) orientations. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Multiphoton ionization processes in strong laser

International Nuclear Information System (INIS)

Krstic, P.

1982-01-01

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

Adaptive optics improves multiphoton super-resolution imaging

Science.gov (United States)

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

2018-02-01

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

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

Science.gov (United States)

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

2010-02-01

High-resolution 3D microscopy based on multiphoton induced autofluorescence and second harmonic generation have been introduced in 1990. 13 years later, CE-marked clinical multiphoton systems for 3D imaging of human skin with subcellular resolution have first been launched by JenLab company with the tomography DermaInspect®. This year, the second generation of clinical multiphoton tomographs was introduced. The novel multiphoton tomograph MPTflex, equipped with a flexible articulated optical arm, provides an increased flexibility and accessibility especially for clinical and cosmetical examinations. Improved image quality and signal to noise ratio (SNR) are achieved by a very short source-drain spacing, by larger active areas of the detectors and by single photon counting (SPC) technology. Shorter image acquisition time due to improved image quality reduces artifacts and simplifies the operation of the system. The compact folded optical design and the light-weight structure of the optical head eases the handling. Dual channel detectors enable to distinguish between intratissue elastic fibers and collagenous structures simultaneously. Through the use of piezo-driven optics a stack of optical cross-sections (optical sectioning) can be acquired and 3D imaging can be performed. The multiphoton excitation of biomolecules like NAD(P)H, flavins, porphyrins, elastin, and melanin is done by picojoule femtosecond laser pulses from an tunable turn-key femtosescond near infrared laser system. The ability for rapid high-quality image acquisition, the user-friendly operation of the system and the compact and flexible design qualifies this system to be used for melanoma detection, diagnostics of dermatological disorders, cosmetic research and skin aging measurements as well as in situ drug monitoring and animal research.

Multiphoton above threshold effects in strong-field fragmentation

DEFF Research Database (Denmark)

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

2012-01-01

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

Advances in polarization sensitive multiphoton nano-bio-imaging

Directory of Open Access Journals (Sweden)

Zyss J.

2010-06-01

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

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

Science.gov (United States)

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

2014-01-01

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

Multiphoton tomography of intratissue tattoo nanoparticles

Science.gov (United States)

König, Karsten

2012-02-01

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

Multiphoton ionization/dissociation of osmium tetroxide

International Nuclear Information System (INIS)

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

1993-01-01

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

Real-time high dynamic range laser scanning microscopy

Science.gov (United States)

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

2016-04-01

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

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

Science.gov (United States)

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

2018-03-25

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

High-energy x-ray microscopy of laser-fusion plasmas at the National Ignition Facility

International Nuclear Information System (INIS)

Koch, J.A.; Landen, O.L.; Hammel, B.A.

1997-01-01

Multi-keV x-ray microscopy will be an important laser-produced plasma diagnostic at future megajoule facilities such as the National Ignition Facility (NIF).In preparation for the construction of this facility, we have investigated several instrumentation options in detail, and we conclude that near normal incidence single spherical or toroidal crystals may offer the best general solution for high-energy x-raymicroscopy at NIF and at similar large facilities. Kirkpatrick-Baez microscopes using multi-layer mirrors may also be good secondary options, particularly if apertures are used to increase the band-width limited field of view

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

International Nuclear Information System (INIS)

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

2004-01-01

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

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

International Nuclear Information System (INIS)

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

2017-01-01

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

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

Science.gov (United States)

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

2004-03-01

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

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

International Nuclear Information System (INIS)

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

2004-01-01

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

Factors associated with malaria microscopy diagnostic performance following a pilot quality-assurance programme in health facilities in malaria low-transmission areas of Kenya, 2014.

Science.gov (United States)

Odhiambo, Fredrick; Buff, Ann M; Moranga, Collins; Moseti, Caroline M; Wesongah, Jesca Okwara; Lowther, Sara A; Arvelo, Wences; Galgalo, Tura; Achia, Thomas O; Roka, Zeinab G; Boru, Waqo; Chepkurui, Lily; Ogutu, Bernhards; Wanja, Elizabeth

2017-09-13

Malaria accounts for ~21% of outpatient visits annually in Kenya; prompt and accurate malaria diagnosis is critical to ensure proper treatment. In 2013, formal malaria microscopy refresher training for microscopists and a pilot quality-assurance (QA) programme for malaria diagnostics were independently implemented to improve malaria microscopy diagnosis in malaria low-transmission areas of Kenya. A study was conducted to identify factors associated with malaria microscopy performance in the same areas. From March to April 2014, a cross-sectional survey was conducted in 42 public health facilities; 21 were QA-pilot facilities. In each facility, 18 malaria thick blood slides archived during January-February 2014 were selected by simple random sampling. Each malaria slide was re-examined by two expert microscopists masked to health-facility results. Expert results were used as the reference for microscopy performance measures. Logistic regression with specific random effects modelling was performed to identify factors associated with accurate malaria microscopy diagnosis. Of 756 malaria slides collected, 204 (27%) were read as positive by health-facility microscopists and 103 (14%) as positive by experts. Overall, 93% of slide results from QA-pilot facilities were concordant with expert reference compared to 77% in non-QA pilot facilities (p malaria diagnosis. Microscopists who had recently completed refresher training and worked in a QA-pilot facility performed the best overall. The QA programme and formal microscopy refresher training should be systematically implemented together to improve parasitological diagnosis of malaria by microscopy in Kenya.

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

International Nuclear Information System (INIS)

Dantus, Marcos

2008-01-01

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

Infrared Multiphoton Dissociation Spectroscopy with Free-Electron Lasers: On the Road from Small Molecules to Biomolecules.

Science.gov (United States)

Jašíková, Lucie; Roithová, Jana

2018-03-07

Infrared multiphoton dissociation (IRMPD) spectroscopy is commonly used to determine the structure of isolated, mass-selected ions in the gas phase. This method has been widely used since it became available at free-electron laser (FEL) user facilities. Thus, in this Minireview, we examine the use of IRMPD/FEL spectroscopy for investigating ions derived from small molecules, metal complexes, organometallic compounds and biorelevant ions. Furthermore, we outline new applications of IRMPD spectroscopy to study biomolecules. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Theory of multiphoton ionization of atoms

International Nuclear Information System (INIS)

Szoeke, A.

1986-03-01

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

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

Science.gov (United States)

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

2017-04-01

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

Pancreatic cancer cell detection by targeted lipid microbubbles and multiphoton imaging

Science.gov (United States)

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

2018-04-01

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

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

International Nuclear Information System (INIS)

Al-azawi, H.R.

1986-01-01

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

Multiphoton tomography to detect chemo- and biohazards

Science.gov (United States)

König, Karsten

2015-03-01

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

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

International Nuclear Information System (INIS)

Ungureanu, C.; Almasan, V.

1994-01-01

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

Multiphoton tomography of the human eye

Science.gov (United States)

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

2017-02-01

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

Preparation of Murine Submandibular Salivary Gland for Upright Intravital Microscopy.

Science.gov (United States)

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

2018-05-07

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

Following Intracellular Cholesterol Transport by Linear and Non-Linear Optical Microscopy of Intrinsically Fluorescent Sterols

DEFF Research Database (Denmark)

Wustner, D.

2012-01-01

Elucidation of intracellular cholesterol transport is important for understanding the molecular basis of several metabolic and neuronal diseases, like atheroclerosis or lysosomal storage disorders. Progress in this field depends crucially on the development of new technical approaches to follow...... is on recent developments in imaging technology to follow the intracellular fate of intrinsically fluorescent sterols as faithful cholesterol markers. In particular, UV-sensitive wide field and multiphoton microscopy of the sterol dehydroergosterol, DHE, is explained and new methods of quantitative image...... analysis like pixel-wise bleach rate fitting and multiphoton image correlation spectroscopy are introduced. Several applications of the new technology including observation of vectorial sterol trafficking in polarized human hepatoma cells for investigation of reverse cholesterol transport are presented....

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

Science.gov (United States)

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

2015-10-01

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

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

Directory of Open Access Journals (Sweden)

Jean-Christophe Pittet

2014-08-01

Full Text Available The aim of this study was to evaluate the influence of the depth of the dermis on the measured collagen and elastin levels and to establish the correlation between the amount of these two extracellular matrix (ECM components and age. Multiphoton Microscopy (MPM that measures the autofluorescence (AF and second harmonic generation (SHG was used to quantify the levels of elastin and collagen and to determine the SAAID (SHG-to-AF Aging Index of Dermis at two different skin depths. A 50 MHz ultrasound scanner was used for the calculation of the Sub Epidermal Non Echogenic Band (SENEB. The measurements of the skin mechanical properties were done with a cutometer. All measurements were performed on two groups of 30 healthy female volunteers. The MPM showed a decrease of the quantity of collagen and elastin as a function of depth of the dermis as well as age. The SAAID was lower for the older skin in the deeper dermis. Ultrasound imaging revealed a significant decrease of SENEB as a function of aging. The mechanical properties confirmed a loss of cutaneous elasticity and firmness. Although multiphoton microscopy is a powerful technique to study the characteristics of the dermis and its age-related damage, the location of the measurements (depth remains very important for the validation of these variations. These variations do not seem to be homogeneous according to the part of the dermis that is studied.

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

Science.gov (United States)

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

2017-02-01

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

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

Science.gov (United States)

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

2017-10-06

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

Three-dimensional spatial imaging in multiphoton ionization rate measurements

International Nuclear Information System (INIS)

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

2004-01-01

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

High-resolution light microscopy using luminescent nanoparticles.

Science.gov (United States)

Ohulchanskyy, Tymish Y; Roy, Indrajit; Yong, Ken-Tye; Pudavar, Haridas E; Prasad, Paras N

2010-01-01

This review presents recent progress in the development of the luminescent nanoparticles for confocal and multiphoton microscopy. Four classes of nanomaterials are discussed: (1) silica-based nanoparticles doped with fluorescent molecules, (2) gold nanoparticles, (3) semiconductor nanocrystals (quantum dots/rods), and (4) nanophosphors. Special considerations are given to recently developed imaging nanoprobes, such as (1) organically modified silica (ORMOSIL) nanoparticles doped with two-photon absorbing fluorophores, which exhibit aggregation-enhanced fluorescence (AEF), and (2) nanophosphors (ceramic nanoparticles containing luminescent lanthanoid ions). Advantages and disadvantages of every class of nanomaterials and their specific applications are briefly discussed.

Advanced Microscopy Facility

Data.gov (United States)

Federal Laboratory Consortium — FUNCTION: Provides a facility for high-resolution studies of complex biomolecular systems. The goal is an understanding of how to engineer biomolecules for various...

Doubly resonant multiphoton ionization

International Nuclear Information System (INIS)

Crance, M.

1978-01-01

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

High-order multiphoton ionization photoelectron spectroscopy of NO

International Nuclear Information System (INIS)

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

1987-01-01

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

Multiphotonic Confocal Microscopy 3D imaging: Application to mantle sulfides in sub-arc environment (Avacha Volcano, Kamchatka)

Science.gov (United States)

Antoine, Bénard; Luc-Serge, Doucet; Sabine, Palle; Dmitri A., Ionov

2010-05-01

Petrogenetic relations in igneous rocks are usually studied in natural samples using classical optical microscopy and subsequent geochemical data acquisition. Multiphotonic Laser Scanning Confocal Microscopy (MLSCM) can be a powerful tool to section geological materials optically with sub-micrometric resolution and then generate a three-dimensional (3D) reconstruction (ca. 106 μm3 stack). MLSCM is used here to investigate textural relations of Monosulfide Solid Solution (MSS) with silicate phases in fresh spinel harzburgite xenoliths from the andesitic Avacha volcano (Kamchatka, Russia). The xenoliths contain MSS disseminated in olivine and orthopyroxene (opx) neoblasts as well as MSS-rich quenched magmatic opx veins [1]. First, Reflection Mode (RM) was tested on vein sulfides in resin-impregnated thick (120 μm) polished rock sections. Then we used a combination of Differential Interference Contrast (DIC) with a transmitted light detector, two photons-excited fluorescence (2PEF) and Second Harmonic Generation (SHG). Sequential imaging feature of the Leica TCS-SP2 software was applied. The excitation laser used for 2PEF was a COHERENT MIRA 900 with a 76Hz repetition rate and 800nm wavelength. Image stacks were analysed using ImageJ software [2]. The aim of the tests was to try to discriminate sulfides in silicate matrix as a tool for a better assessment of equilibrium conditions between the two phases. Preliminary results show that Fe-Ni rich MSS from vein and host rock have a strong auto-fluorescence in the Near UV-VIS domain (392-715 nm) whereas silicate matrix is only revealed through DIC. SHG is obtained only from dense nanocentrosymmetrical structures such as embedded medium (organic matter like glue and resin). The three images were recorded sequentially enabling efficient discrimination between the different components of the rock slices. RM permits reconstruction of the complete 3D structure of the rock slice. High resolution (ca. 0.2 μm along X-Y axis vs

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

International Nuclear Information System (INIS)

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

2009-01-01

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

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

International Nuclear Information System (INIS)

Yu, An

1997-01-01

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

Efficient Multiphoton Generation in Waveguide Quantum Electrodynamics

Science.gov (United States)

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

2017-05-01

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

Multiphoton processes in isolated atoms and molecules

International Nuclear Information System (INIS)

Sudbo, A.S.

1979-11-01

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

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

Science.gov (United States)

Watson, Jennifer M.

Ovarian cancer is a deadly disease owing to the non-specific symptoms and suspected rapid progression, leading to frequent late stage detection and poor prognosis. Medical imaging methods such as CT, MRI and ultrasound as well as serum testing for cancer markers have had extremely poor performance for early disease detection. Due to the poor performance of available screening methods, and the impracticality and ineffectiveness of taking tissue biopsies from the ovary, women at high risk for developing ovarian cancer are often advised to undergo prophylactic salpingo-oophorectomy. This surgery results in many side effects and is most often unnecessary since only a fraction of high risk women go on to develop ovarian cancer. Better understanding of the early development of ovarian cancer and characterization of morphological changes associated with early disease could lead to the development of an effective screening test for women at high risk. Optical imaging methods including optical coherence tomography (OCT) and multiphoton microscopy (MPM) are excellent tools for studying disease progression owing to the high resolution and depth sectioning capabilities. Further, these techniques are excellent for optical biopsy because they can image in situ non-destructively. In the studies described in this dissertation OCT and MPM are used to identify cellular and tissue morphological changes associated with early tumor development in a mouse model of ovarian cancer. This work is organized into three specific aims. The first aim is to use the images from the MPM phenomenon of second harmonic generation to quantitatively examine the morphological differences in collagen structure in normal mouse ovarian tissue and mouse ovarian tumors. The second aim is to examine the differences in endogenous two-photon excited fluorescence in normal mouse ovarian tissue and mouse ovarian tumors. The third and final aim is to identify changes in ovarian microstructure resulting from early

Additive Manufacture of Three Dimensional Nanocomposite Based Objects through Multiphoton Fabrication

Directory of Open Access Journals (Sweden)

Yaan Liu

2016-09-01

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

Fundamentals of fluorescence microscopy exploring life with light

CERN Document Server

Mondal, Partha Pratim

2014-01-01

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

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

International Nuclear Information System (INIS)

Takeuchi, Kazuo; Nakane, Ryohei; Inoue, Cihiro

1981-01-01

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

Advanced light microscopy core facilities: Balancing service, science and career

Science.gov (United States)

Hartmann, Hella; Reymann, Jürgen; Ansari, Nariman; Utz, Nadine; Fried, Hans‐Ulrich; Kukat, Christian; Peychl, Jan; Liebig, Christian; Terjung, Stefan; Laketa, Vibor; Sporbert, Anje; Weidtkamp‐Peters, Stefanie; Schauss, Astrid; Zuschratter, Werner; Avilov, Sergiy

2016-01-01

ABSTRACT Core Facilities (CF) for advanced light microscopy (ALM) have become indispensable support units for research in the life sciences. Their organizational structure and technical characteristics are quite diverse, although the tasks they pursue and the services they offer are similar. Therefore, throughout Europe, scientists from ALM‐CFs are forming networks to promote interactions and discuss best practice models. Here, we present recommendations for ALM‐CF operations elaborated by the workgroups of the German network of ALM‐CFs, German Bio‐Imaging (GerBI). We address technical aspects of CF planning and instrument maintainance, give advice on the organization and management of an ALM‐CF, propose a scheme for the training of CF users, and provide an overview of current resources for image processing and analysis. Further, we elaborate on the new challenges and opportunities for professional development and careers created by CFs. While some information specifically refers to the German academic system, most of the content of this article is of general interest for CFs in the life sciences. Microsc. Res. Tech. 79:463–479, 2016. © 2016 THE AUTHORS MICROSCOPY RESEARCH AND TECHNIQUE PUBLISHED BY WILEY PERIODICALS, INC. PMID:27040755

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

International Nuclear Information System (INIS)

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

1999-01-01

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

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

Science.gov (United States)

Seemann, K M; Kuhn, B

2014-07-01

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

Dressing effect in multiphoton unimolecular dissociation

Energy Technology Data Exchange (ETDEWEB)

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

1986-03-01

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

National Cryo-Electron Microscopy Facility

Science.gov (United States)

Information about the National Cryo-EM Facility at NCI, created to provide researchers access to the latest cryo-EM technology for high resolution imaging. Includes timeline for installation and how to access the facility.

Brain morphology imaging by 3D microscopy and fluorescent Nissl staining.

Science.gov (United States)

Lazutkin, A A; Komissarova, N V; Toptunov, D M; Anokhin, K V

2013-07-01

Modern optical methods (multiphoton and light-sheet fluorescent microscopy) allow 3D imaging of large specimens of the brain with cell resolution. It is therefore essential to refer the resultant 3D pictures of expression of transgene, protein, and other markers in the brain to the corresponding structures in the atlas. This implies counterstaining of specimens with morphological dyes. However, there are no methods for contrasting large samples of the brain without their preliminary slicing. We have developed a method for fluorescent Nissl staining of whole brain samples. 3D reconstructions of specimens of the hippocampus, olfactory bulbs, and cortex were created. The method can be used for morphological control and evaluation of the effects of various factors on the brain using 3D microscopy technique.

Generalized Multiphoton Quantum Interference

Directory of Open Access Journals (Sweden)

Max Tillmann

2015-10-01

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

Rapid in vivo vertical tissue sectioning by multiphoton tomography

Science.gov (United States)

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

2018-02-01

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

Multiphoton microscopy of ECM proteins in baboon aortic leaflet

Science.gov (United States)

Gonzalez, Mariacarla; Saytashev, Ilyas; Luna, Camila; Gonzalez, Brittany; Pinero, Alejandro; Perez, Manuel; Ramaswamy, Sharan; Ramella-Roman, Jessica

2018-02-01

The extracellular matrix (ECM) plays crucial role in defining mechanical properties of a heart valve yet the mechanobiological role of the ECM proteins - collagen and elastin - in living heart valve leaflets is still poorly understood. In this study, non-linear microscopy was used to obtain three dimensional images of collagen and elastin arrangement in aortic leaflets under combined steady flow (850 ml/min) and cyclic flexure (1 Hz) mechanical (dynamic) training. A novel bioreactor capable of mimicking the flow conditions in a living heart was used in this study and was optimized for microscopic imagery. A custom made non-linear microscope was used in this study to provide Second Harmonic Generation (SHG) imaging of collagen arrangement and two-photon imaging of elastin. Two control and three trained leaflet samples from static and dynamic tissue culture were imaged to observe protein changes in the tissue for a period of seven days. Dynamic training led to a decrease in alignment index of the protein fibers compared to the static treatment.

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

Science.gov (United States)

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

2004-03-01

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

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

International Nuclear Information System (INIS)

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

2004-01-01

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

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

International Nuclear Information System (INIS)

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

2015-01-01

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

Invited Review Article: Pump-probe microscopy

Science.gov (United States)

Wilson, Jesse W.; Robles, Francisco E.; Warren, Warren S.

2016-01-01

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

Invited Review Article: Pump-probe microscopy

Energy Technology Data Exchange (ETDEWEB)

Fischer, Martin C., E-mail: Martin.Fischer@duke.edu; Wilson, Jesse W.; Robles, Francisco E. [Department of Chemistry, Duke University, Durham, North Carolina 27708 (United States); Warren, Warren S. [Departments of Chemistry, Biomedical Engineering, Physics, and Radiology, Duke University, Durham, North Carolina 27708 (United States)

2016-03-15

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

Fundamental studies of molecular multiphoton ionization

International Nuclear Information System (INIS)

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

1984-04-01

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

Multiphoton gonioscopy to image the trabecular meshwork of porcine eyes

Science.gov (United States)

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

2013-03-01

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

Direct trabecular meshwork imaging in porcine eyes through multiphoton gonioscopy

Science.gov (United States)

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

2013-03-01

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

Label-free imaging of acanthamoeba using multimodal nonlinear optical microscopy

Science.gov (United States)

Kobayashi, Tsubasa; Cha, Yu-Rok; Kaji, Yuichi; Oshika, Tetsuro; Leproux, Philippe; Couderc, Vincent; Kano, Hideaki

2018-02-01

Acanthamoeba keratitis is a disease in which amoebae named Acanthamoeba invade the cornea of an eye. To diagnose this disease before it becomes serious, it is important to detect the cyst state of Acanthamoeba in the early stage of infection. In the present study, we explored spectroscopic signitures of the cyst state of Acanthamoeba using multimodal nonlinear optical microscopy with the channels of multiplex coherent anti-Stokes Raman scattering (CARS), second harmonic generation (SHG), and third harmonic generation (THG). A sharp band at around 1603 cm-1 in the CARS (Im[χ(3)]) spectrum was found at the cyst state of Acanthamoeba, which possibly originates from ergosterol and/or 7-dehydrostigmasterol. It can be used as a maker band of Acanthamoeba for medical treatment. Keyword: Acanthamoeba keratitis, coherent anti-Stokes Raman scattering, CARS, second harmonic generation, SHG, microspectroscopy, multiphoton microscopy

Multiphoton dissociation of polyatomic molecules

International Nuclear Information System (INIS)

Schulz, P.A.

1979-10-01

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

Multiphoton photodegradation of indocyanine green: Solvent protolysis effect

Energy Technology Data Exchange (ETDEWEB)

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

2016-02-15

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

Influence of rotation on multiphoton processes in HF

International Nuclear Information System (INIS)

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

1994-01-01

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

Investigation of the effect of hydration on dermal collagen in ex vivo human skin tissue using second harmonic generation microscopy

Science.gov (United States)

Samatham, Ravikant; Wang, Nicholas K.; Jacques, Steven L.

2016-02-01

Effect of hydration on the dermal collagen structure in human skin was investigated using second harmonic generation microscopy. Dog ears from the Mohs micrographic surgery department were procured for the study. Skin samples with subject aged between 58-90 years old were used in the study. Three dimensional Multiphoton (Two-photon and backward SHG) control data was acquired from the skin samples. After the control measurement, the skin tissue was either soaked in deionized water for 2 hours (Hydration) or kept at room temperature for 2 hours (Desiccation), and SHG data was acquired. The data was normalized for changes in laser power and detector gain. The collagen signal per unit volume from the dermis was calculated. The desiccated skin tissue gave higher backward SHG compared to respective control tissue, while hydration sample gave a lower backward SHG. The collagen signal decreased with increase in hydration of the dermal collagen. Hydration affected the packing of the collagen fibrils causing a change in the backward SHG signal. In this study, the use of multiphoton microscopy to study the effect of hydration on dermal structure was demonstrated in ex vivo tissue.

Ponderomotive effects in multiphoton pair production

Science.gov (United States)

Kohlfürst, Christian; Alkofer, Reinhard

2018-02-01

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

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

International Nuclear Information System (INIS)

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

2007-01-01

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

Nonlinear excitation fluorescence microscopy: source considerations for biological applications

Science.gov (United States)

Wokosin, David L.

2008-02-01

Ultra-short-pulse solid-state laser sources have improved contrast within fluorescence imaging and also opened new windows of investigation in biological imaging applications. Additionally, the pulsed illumination enables harmonic scattering microscopy which yields intrinsic structure, symmetry and contrast from viable embryos, cells and tissues. Numerous human diseases are being investigated by the combination of (more) intact dynamic tissue imaging of cellular function with gene-targeted specificity and electrophysiology context. The major limitation to more widespread use of multi-photon microscopy has been the complete system cost and added complexity above and beyond commercial camera and confocal systems. The current status of all-solid-state ultrafast lasers as excitation sources will be reviewed since these lasers offer tremendous potential for affordable, reliable, "turnkey" multiphoton imaging systems. This effort highlights the single box laser systems currently commercially available, with defined suggestions for the ranges for individual laser parameters as derived from a biological and fluorophore limited perspective. The standard two-photon dose is defined by 800nm, 10mW, 200fs, and 80Mhz - at the sample plane for tissue culture cells, i.e. after the full scanning microscope system. Selected application-derived excitation wavelengths are well represented by 700nm, 780nm, ~830nm, ~960nm, 1050nm, and 1250nm. Many of the one-box lasers have fixed or very limited excitation wavelengths available, so the lasers will be lumped near 780nm, 800nm, 900nm, 1050nm, and 1250nm. The following laser parameter ranges are discussed: average power from 200mW to 2W, pulse duration from 70fs to 700fs, pulse repetition rate from 20MHz to 200MHz, with the laser output linearly polarized with an extinction ratio at least 100:1.

Brain plasticity and functionality explored by nonlinear optical microscopy

Science.gov (United States)

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

2010-02-01

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

Superficial nephrons in BALB/c and C57BL/6 mice facilitate in vivo multiphoton microscopy of the kidney.

Directory of Open Access Journals (Sweden)

Ina Maria Schießl

Full Text Available Multiphoton microscopy (MPM offers a unique approach for addressing both the function and structure of an organ in near-real time in the live animal. The method however is limited by the tissue-specific penetration depth of the excitation laser. In the kidney, structures in the range of 100 µm from the surface are accessible for MPM. This limitation of MPM aggravates the investigation of the function of structures located deeper in the renal cortex, like the glomerulus and the juxtaglomerular apparatus. In view of the relevance of gene-targeted mice for investigating the function of these structures, we aimed to identify a mouse strain with a high percentage of superficially located glomeruli. The mean distance of the 30 most superficial glomeruli from the kidney surface was determined in 10 commonly used mouse strains. The mean depth of glomeruli was 118.4±3.4, 123.0±2.7, 133.7±3.0, 132.3±2.6, 141.0±4.0, 145.3±4.3, 148.9±4.2, 151.6±2.7, 167.7±3.9, and 207.8±3.2 µm in kidney sections from 4-week-old C3H/HeN, BALB/cAnN, SJL/J, C57BL/6N, DBA/2N, CD1 (CRI, 129S2/SvPas, CB6F1, FVB/N and NMRI (Han mice, respectively (n = 5 animals from each strain. The mean distance from the kidney surface of the most superficial glomeruli was significantly lower in the strains C3H/HeN Crl, BALB/cAnN, DBA/2NCrl, and C57BL/6N when compared to a peer group consisting of all the other strains (p<.0001. In 10-week-old mice, the most superficial glomeruli were located deeper in the cortex when compared to 4-week-old animals, with BALB/cAnN and C57BL/6N being the strains with the highest percentage of superficial glomeruli (25% percentile 116.7 and 121.9 µm, respectively. In summary, due to significantly more superficial glomeruli compared to other commonly used strains, BALB/cAnN and C57BL/6N mice appear to be particularly suitable for the investigation of glomerular function using MPM.

Rapid volumetric imaging with Bessel-Beam three-photon microscopy

Science.gov (United States)

Chen, Bingying; Huang, Xiaoshuai; Gou, Dongzhou; Zeng, Jianzhi; Chen, Guoqing; Pang, Meijun; Hu, Yanhui; Zhao, Zhe; Zhang, Yunfeng; Zhou, Zhuan; Wu, Haitao; Cheng, Heping; Zhang, Zhigang; Xu, Chris; Li, Yulong; Chen, Liangyi; Wang, Aimin

2018-01-01

Owing to its tissue-penetration ability, multi-photon fluorescence microscopy allows for the high-resolution, non-invasive imaging of deep tissue in vivo; the recently developed three-photon microscopy (3PM) has extended the depth of high-resolution, non-invasive functional imaging of mouse brains to beyond 1.0 mm. However, the low repetition rate of femtosecond lasers that are normally used in 3PM limits the temporal resolution of point-scanning three-photon microscopy. To increase the volumetric imaging speed of 3PM, we propose a combination of an axially elongated needle-like Bessel-beam with three-photon excitation (3PE) to image biological samples with an extended depth of focus. We demonstrate the higher signal-to-background ratio (SBR) of the Bessel-beam 3PM compared to the two-photon version both theoretically and experimentally. Finally, we perform simultaneous calcium imaging of brain regions at different axial locations in live fruit flies and rapid volumetric imaging of neuronal structures in live mouse brains. These results highlight the unique advantage of conducting rapid volumetric imaging with a high SBR in the deep brain in vivo using scanning Bessel-3PM.

Resonance Enhanced Multi-photon Spectroscopy of DNA

Science.gov (United States)

Ligare, Marshall Robert

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

Slide-free histology via MUSE: UV surface excitation microscopy for imaging unsectioned tissue (Conference Presentation)

Science.gov (United States)

Levenson, Richard M.; Harmany, Zachary; Demos, Stavros G.; Fereidouni, Farzad

2016-03-01

Widely used methods for preparing and viewing tissue specimens at microscopic resolution have not changed for over a century. They provide high-quality images but can involve time-frames of hours or even weeks, depending on logistics. There is increasing interest in slide-free methods for rapid tissue analysis that can both decrease turn-around times and reduce costs. One new approach is MUSE (microscopy with UV surface excitation), which exploits the shallow penetration of UV light to excite fluorescent signals from only the most superficial tissue elements. The method is non-destructive, and eliminates requirement for conventional histology processing, formalin fixation, paraffin embedding, or thin sectioning. It requires no lasers, confocal, multiphoton or optical coherence tomography optics. MUSE generates diagnostic-quality histological images that can be rendered to resemble conventional hematoxylin- and eosin-stained samples, with enhanced topographical information, from fresh or fixed, but unsectioned tissue, rapidly, with high resolution, simply and inexpensively. We anticipate that there could be widespread adoption in research facilities, hospital-based and stand-alone clinical settings, in local or regional pathology labs, as well as in low-resource environments.

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

Science.gov (United States)

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

2017-10-01

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

Multiphoton spectroscopy of human skin in vivo

Science.gov (United States)

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

2012-03-01

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

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

Energy Technology Data Exchange (ETDEWEB)

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

2010-11-15

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

Quantitative multiphoton imaging

Science.gov (United States)

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

2014-02-01

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

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

International Nuclear Information System (INIS)

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

2002-01-01

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

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

International Nuclear Information System (INIS)

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

1986-01-01

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

External quality assessment of AFB smear microscopy performances and its associated factors in selected private health facilities in Addis Ababa, Ethiopia.

Science.gov (United States)

Mosissa, Lemi; Kebede, Abebaw; Mindaye, Tedla; Getahun, Muluwork; Tulu, Sisay; Desta, Kassu

2016-01-01

Tuberculosis (TB) is still a public health problem in sub Saharan African countries. In resource-limited settings, TB diagnosis relies on sputum smear microscopy, with low and variable sensitivities, especially in paucibacillary pediatric and HIV-associated TB patients. Tuberculosis microscopy centers have several weaknesses like overworking, insufficiently trained personnel, inconsistent reagent supplies, and poorly maintained equipments; thus, there is a critical need for investments in laboratory infrastructure, capacity building, and quality assurance schemes. The performance of TB microscopy centers in the private health facilities in Addis Ababa is not known so far. The main objective of the study was to assess laboratory performance of acid fast bacilli (AFB) smear microscopy and its associated factors in selected private health facilities in Addis Ababa, Ethiopia. A cross-sectional study was conducted in 33 selected private health facilities of Addis Ababa, Ethiopia comprising 7 hospitals, 2 NGO health centers, 23 higher clinics and 1 diagnostic laboratory that provide AFB smear microscopy services. The study was conducted from January to April 2014. A total of 283 stained sputum smears were randomly collected from participant laboratories for blinded rechecking, 320 panel slides were sent to 32 microscopy centers to evaluate their performance on AFB reading, staining and reporting. Checklists were used to assess quality issues of laboratories. Data were captured, cleaned, and analyzed using SPSS version 16.0; χ(2) tests, kappa statistics were used for comparison purpose. P value laboratories, 2-scored 100%, 15 scored 80-95% & the remaining 15 scored 50-75% for overall proficiency test performance. There were 10 (3.15%) major errors and 121 (37.8%) minor errors. The sensitivity, specificity, PPV and NPV of panel reading by microscopy centers were 89%, 96%, 96%, and 90% respectively. Out of 283 randomly selected slides for blind rechecking, 11 (3.9%) slides

Graphene oxide from silk cocoon: a novel magnetic fluorophore for multi-photon imaging.

Science.gov (United States)

Roy, Manas; Kusurkar, Tejas Sanjeev; Maurya, Sandeep Kumar; Meena, Sunil Kumar; Singh, Sushil Kumar; Sethy, Niroj; Bhargava, Kalpana; Sharma, Raj Kishore; Goswami, Debabrata; Sarkar, Sabyasachi; Das, Mainak

2014-02-01

In this work, we synthesized graphene oxide from silk cocoon embarking its new dimension as a magnetic fluorophore when compared with its present technical status, which at best is for extracting silk as a biomaterial for tissue engineering applications. We produced graphene oxide by pyrolysing the silk cocoon in an inert atmosphere. The collected raw carbon is oxidized by nitric acid that readily produces multilayer graphene oxide with nano carbon particulates. Structural properties of the graphene oxide were analyzed using scanning electron microscopy, transmission electron microscopy, Fourier transform infra-red spectroscopy, and Raman spectroscopy. The oxidized sample shows remarkable fluorescence, multi-photon imaging and magnetic properties. On increasing the excitation wavelength, the fluorescence emission intensity of the graphene oxide also increases and found maximum emission at 380 nm excitation wavelength. On studying the two photon absorption (TPA) property of aqueous graphene oxide using Z-scan technique, we found significant TPA activity at near infrared wavelength. In addition, the graphene oxide shows ferromagnetic behavior at room temperature. The observed fluorescence and magnetic property were attributed to the defects caused in the graphene oxide structure by introducing oxygen containing hydrophilic groups during the oxidation process. Previously silk cocoon has been used extensively in deriving silk-based tissue engineering materials and as gas filter. Here we show a novel application of silk cocoon by synthesizing graphene oxide based magnetic-fluorophore for bio-imaging applications.

Multiphoton absorption coefficients in solids: an universal curve

International Nuclear Information System (INIS)

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

1983-04-01

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

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

Czech Academy of Sciences Publication Activity Database

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

2006-01-01

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

Multiphoton microscopic imaging of histological sections without hematoxylin and eosin staining differentiates carcinoma in situ lesion from normal oesophagus

Science.gov (United States)

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

2013-10-01

Multiphoton microscopy (MPM) has become a powerful, important tool for tissues imaging at the molecular level. In this paper, this technique was extended to histological investigations, differentiating carcinoma in situ (CIS) lesion from normal oesophagus by imaging histological sections without hematoxylin and eosin (H&E) staining. The results show that the histology procedures of dehydration, paraffin embedding, and de-paraffinizing highlighted two photon excited fluorescence of cytoplasm and nucleolus of epithelial cell and collagen in stroma. MPM has the ability to identify the characteristics of CIS lesion including changes of squamous cells and full epithelium, identification of basement membrane, especially prominent nucleolus. The studies described here show that MPM has the potential for future retrospective studies of tumor staging by employing on histological section specimens without H&E staining.

Multiphoton minimal inertia scanning for fast acquisition of neural activity signals

Science.gov (United States)

Schuck, Renaud; Go, Mary Ann; Garasto, Stefania; Reynolds, Stephanie; Dragotti, Pier Luigi; Schultz, Simon R.

2018-04-01

Objective. Multi-photon laser scanning microscopy provides a powerful tool for monitoring the spatiotemporal dynamics of neural circuit activity. It is, however, intrinsically a point scanning technique. Standard raster scanning enables imaging at subcellular resolution; however, acquisition rates are limited by the size of the field of view to be scanned. Recently developed scanning strategies such as travelling salesman scanning (TSS) have been developed to maximize cellular sampling rate by scanning only select regions in the field of view corresponding to locations of interest such as somata. However, such strategies are not optimized for the mechanical properties of galvanometric scanners. We thus aimed to develop a new scanning algorithm which produces minimal inertia trajectories, and compare its performance with existing scanning algorithms. Approach. We describe here the adaptive spiral scanning (SSA) algorithm, which fits a set of near-circular trajectories to the cellular distribution to avoid inertial drifts of galvanometer position. We compare its performance to raster scanning and TSS in terms of cellular sampling frequency and signal-to-noise ratio (SNR). Main Results. Using surrogate neuron spatial position data, we show that SSA acquisition rates are an order of magnitude higher than those for raster scanning and generally exceed those achieved by TSS for neural densities comparable with those found in the cortex. We show that this result also holds true for in vitro hippocampal mouse brain slices bath loaded with the synthetic calcium dye Cal-520 AM. The ability of TSS to ‘park’ the laser on each neuron along the scanning trajectory, however, enables higher SNR than SSA when all targets are precisely scanned. Raster scanning has the highest SNR but at a substantial cost in number of cells scanned. To understand the impact of sampling rate and SNR on functional calcium imaging, we used the Cramér-Rao Bound on evoked calcium traces recorded

Line broadening in multiphoton processes with a resonant intermediate transition

International Nuclear Information System (INIS)

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

1983-01-01

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

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

International Nuclear Information System (INIS)

Popov, V.S.

2001-01-01

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

Femtosecond Light Source for Phase-Controlled Multiphoton Ionization

International Nuclear Information System (INIS)

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

2001-01-01

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

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

Directory of Open Access Journals (Sweden)

Stratos Galanopoulos

2014-08-01

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

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

International Nuclear Information System (INIS)

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

2004-01-01

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

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

Science.gov (United States)

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

2004-02-01

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

Coherent enhancement of resonance-mediated multiphoton absorption

International Nuclear Information System (INIS)

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

2010-01-01

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

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

International Nuclear Information System (INIS)

Rockney, B.H.

1982-01-01

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

NICHD Microscopy and Imaging Core (MIC)

Data.gov (United States)

Federal Laboratory Consortium — The NICHD Microscopy and Imaging Core (MIC) is designed as a multi-user research facility providing training and instrumentation for high resolution microscopy and...

Utilizing two-photon fluorescence and second harmonic generation microscopy to study human bone marrow mesenchymal stem cell morphogenesis in chitosan scaffold

Science.gov (United States)

Su, Ping-Jung; Huang, Chi-Hsiu; Huang, Yi-You; Lee, Hsuan-Sue; Dong, Chen-Yuan

2008-02-01

A major goal of tissue engineering is to cultivate the cartilage in vitro. One approach is to implant the human bone marrow mesenchymal stem cells into the three dimensional biocompatible and biodegradable material. Through the action of the chondrogenic factor TGF-β3, the stem cells can be induced to secrete collagen. In this study, mesenchymal stem cells are implanted on the chitosan scaffold and TGF-β3 was added to produce the cartilage tissue and TP autofluorescence and SHG microscopy was used to image the process of chondrogenesis. With additional development, multiphoton microscopy can be developed into an effective tool for evaluating the quality of tissue engineering products.

X-ray microscopy in Aarhus

International Nuclear Information System (INIS)

Uggerhoej, Erik; Abraham-Peskir, Joanna V.

2000-01-01

The Aarhus imaging soft X-ray microscope is now a busy multi-user facility. The optical set-up will be described and project highlights discussed. a) Metal-induced structural changes in whole cells in solution. The effects of aluminum, copper, nickel and zinc on protozoa investigated by using a combination of light microscopy, confocal scanning laser microscopy and X-ray microscopy. b) Botanical studies by X-ray microscopy used to compliment electron microscopy studies. c) Sludge morphology and iron precipitation in Danish freshwater plants by combining X-ray, scanning electron and transmission electron microscopy

Real-time histology in liver disease using multiphoton microscopy with fluorescence lifetime imaging

OpenAIRE

Wang, Haolu; Liang, Xiaowen; Mohammed, Yousuf H.; Thomas, James A.; Bridle, Kim R.; Thorling, Camilla A.; Grice, Jeffrey E.; Xu, Zhi Ping; Liu, Xin; Crawford, Darrell H. G.; Roberts, Michael S.

2015-01-01

Conventional histology with light microscopy is essential in the diagnosis of most liver diseases. Recently, a concept of real-time histology with optical biopsy has been advocated. In this study, live mice livers (normal, with fibrosis, steatosis, hepatocellular carcinoma and ischemia-reperfusion injury) were imaged by MPM-FLIM for stain-free real-time histology. The acquired MPM-FLIM images were compared with conventional histological images. MPM-FLIM imaged subsurface cellular and subcellu...

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

International Nuclear Information System (INIS)

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

2007-01-01

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

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

International Nuclear Information System (INIS)

Prepelitsa, O.B.

1999-01-01

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

vuv fluorescence from selective high-order multiphoton excitation of N2

International Nuclear Information System (INIS)

Coffee, Ryan N.; Gibson, George N.

2004-01-01

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

Summary of 2016 Light Microscopy Module (LMM) Physical Science Experiments on ISS. Update of LMM Science Experiments and Facility Capabilities

Science.gov (United States)

Sicker, Ronald J.; Meyer, William V.; Foster, William M.; Fletcher, William A.; Williams, Stuart J.; Lee, Chang-Soo

2016-01-01

This presentation will feature a series of short, entertaining, and informative videos that describe the current status and science support for the Light Microscopy Module (LMM) facility on the International Space Station. These interviews will focus on current experiments and provide an overview of future capabilities. The recently completed experiments include nano-particle haloing, 3-D self-assembly with Janus particles and a model system for nano-particle drug delivery. The videos will share perspectives from the scientists, engineers, and managers working with the NASA Light Microscopy program.

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

Energy Technology Data Exchange (ETDEWEB)

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

1965-07-01

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

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

International Nuclear Information System (INIS)

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

1990-01-01

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

Investigations of multiphoton excitation and ionization in a short range potential

International Nuclear Information System (INIS)

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

1989-02-01

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

Investigations of multiphoton excitation and ionization in a short range potential

Energy Technology Data Exchange (ETDEWEB)

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

1989-02-01

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

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

Science.gov (United States)

Tong, Ming Hui; Huang, Nan; Zhang, Wei; Zhou, Zhuo Long; Ngan, Alfonso Hing Wan; Du, Yanan; Chan, Barbara Pui

2016-01-01

Engineering 3D microstructures with predetermined properties is critical for stem cell niche studies. We have developed a multiphoton femtosecond laser-based 3D printing platform, which generates complex protein microstructures in minutes. Here, we used the platform to test a series of fabrication and reagent parameters in precisely controlling the mechanical properties of protein micropillars. Atomic force microscopy was utilized to measure the reduced elastic modulus of the micropillars, and transmission electron microscopy was used to visualize the porosity of the structures. The reduced elastic modulus of the micropillars associated positively and linearly with the scanning power. On the other hand, the porosity and pore size of the micropillars associated inversely and linearly with the scanning power and reagent concentrations. While keeping the elastic modulus constant, the stiffness of the micropillars was controlled by varying their height. Subsequently, the single cell traction forces of rabbit chondrocytes, human dermal fibroblasts, human mesenchymal stem cells, and bovine nucleus pulposus cells (bNPCs) were successfully measured by culturing the cells on micropillar arrays of different stiffness. Our results showed that the traction forces of all groups showed positive relationship with stiffness, and that the chondrocytes and bNPCs generated the highest and lowest traction forces, respectively.

Non-perturbative methods applied to multiphoton ionization

International Nuclear Information System (INIS)

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

1982-09-01

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

Wavefront optimized nonlinear microscopy of ex vivo human retinas

Science.gov (United States)

Gualda, Emilio J.; Bueno, Juan M.; Artal, Pablo

2010-03-01

A multiphoton microscope incorporating a Hartmann-Shack (HS) wavefront sensor to control the ultrafast laser beam's wavefront aberrations has been developed. This instrument allowed us to investigate the impact of the laser beam aberrations on two-photon autofluorescence imaging of human retinal tissues. We demonstrated that nonlinear microscopy images are improved when laser beam aberrations are minimized by realigning the laser system cavity while wavefront controlling. Nonlinear signals from several human retinal anatomical features have been detected for the first time, without the need of fixation or staining procedures. Beyond the improved image quality, this approach reduces the required excitation power levels, minimizing the side effects of phototoxicity within the imaged sample. In particular, this may be important to study the physiology and function of the healthy and diseased retina.

In vivo pump-probe microscopy of melanoma and pigmented lesions

Science.gov (United States)

Wilson, Jesse W.; Degan, Simone; Mitropoulos, Tanya; Selim, M. Angelica; Zhang, Jennifer Y.; Warren, Warren S.

2012-03-01

A growing number of dermatologists and pathologists are concerned that the rapidly rising incidence of melanoma reflects not a true 'epidemic' but an increasing tendency to overdiagnose pigmented lesions. Addressing this problem requires both a better understanding of early-stage melanoma and new diagnostic criteria based on more than just cellular morphology and architecture. Here we present a method for in-vivo optical microscopy that utilizes pump-probe spectroscopy to image the distribution of the two forms of melanin in skin: eumelanin and pheomelanin. Images are acquired in a scanning microscope with a sensitive modulation transfer technique by analyzing back-scattered probe light with a lock-in amplifier. Early-stage melanoma is studied in a human skin xenografted mouse model. Individual melanocytes have been observed, in addition to pigmented keratinocytes. Combining the pump-probe images simultaneously with other noninvasive laser microscopy methods (confocal reflectance, multiphoton autofluorescence, and second harmonic generation) allows visualization of the skin architecture, framing the functional pump-probe image in the context of the surrounding tissue morphology. It is found that pump-probe images of melanin can be acquired with low peak intensities, enabling wide field-of-view pigmentation surveys. Finally, we investigate the diagnostic potential of the additional chemical information available from pump-probe microscopy.

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

Energy Technology Data Exchange (ETDEWEB)

Yokoya, Kaoru; Chen, Pisin

1989-03-01

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

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

Science.gov (United States)

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

2003-05-01

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

Neural plasticity explored by correlative two-photon and electron/SPIM microscopy

Science.gov (United States)

Allegra Mascaro, A. L.; Silvestri, L.; Costantini, I.; Sacconi, L.; Maco, B.; Knott, G. W.; Pavone, F. S.

2013-06-01

Plasticity of the central nervous system is a complex process which involves the remodeling of neuronal processes and synaptic contacts. However, a single imaging technique can reveal only a small part of this complex machinery. To obtain a more complete view, complementary approaches should be combined. Two-photon fluorescence microscopy, combined with multi-photon laser nanosurgery, allow following the real-time dynamics of single neuronal processes in the cerebral cortex of living mice. The structural rearrangement elicited by this highly confined paradigm of injury can be imaged in vivo first, and then the same neuron could be retrieved ex-vivo and characterized in terms of ultrastructural features of the damaged neuronal branch by means of electron microscopy. Afterwards, we describe a method to integrate data from in vivo two-photon fluorescence imaging and ex vivo light sheet microscopy, based on the use of major blood vessels as reference chart. We show how the apical dendritic arbor of a single cortical pyramidal neuron imaged in living mice can be found in the large-scale brain reconstruction obtained with light sheet microscopy. Starting from its apical portion, the whole pyramidal neuron can then be segmented and located in the correct cortical layer. With the correlative approach presented here, researchers will be able to place in a three-dimensional anatomic context the neurons whose dynamics have been observed with high detail in vivo.

Graphene-enabled electron microscopy and correlated super-resolution microscopy of wet cells.

Science.gov (United States)

Wojcik, Michal; Hauser, Margaret; Li, Wan; Moon, Seonah; Xu, Ke

2015-06-11

The application of electron microscopy to hydrated biological samples has been limited by high-vacuum operating conditions. Traditional methods utilize harsh and laborious sample dehydration procedures, often leading to structural artefacts and creating difficulties for correlating results with high-resolution fluorescence microscopy. Here, we utilize graphene, a single-atom-thick carbon meshwork, as the thinnest possible impermeable and conductive membrane to protect animal cells from vacuum, thus enabling high-resolution electron microscopy of wet and untreated whole cells with exceptional ease. Our approach further allows for facile correlative super-resolution and electron microscopy of wet cells directly on the culturing substrate. In particular, individual cytoskeletal actin filaments are resolved in hydrated samples through electron microscopy and well correlated with super-resolution results.

Controlling the optical bistability beyond the multi-photon resonance condition in a three-level closed-loop atomic system

International Nuclear Information System (INIS)

Mahmoudi, Mohammad; Nozari, Narges; Vafafard, Azar; Sahrai, Mostafa

2012-01-01

We investigate the optical bistability behavior of a three-level closed-loop atomic system beyond the multi-photon resonance condition. Using the Floquet decomposition, we solve the time-dependent equations of motion, beyond the multi-photon resonance condition. By identifying the different scattering processes contributing to the medium response, it is shown that in general the optical bistability behavior of the system is not phase-dependent. The phase dependence is due to the scattering of the driving and coupling fields into the probe field at a frequency, which, in general, differs from the probe field frequency. - Highlights: → We investigate optical bistability of a three-level closed-loop atomic system, beyond the multi-photon resonance condition. → By applying Floquet decomposition to the equation of motion, the different scattering processes contributing to the medium response are determined. → It is shown that the phase dependence of optical bistability arises from the scattering of the driving and coupling fields into the probe field frequency.

Photo-induced processes in collagen-hypericin system revealed by fluorescence spectroscopy and multiphoton microscopy

OpenAIRE

Hovhannisyan, V.; Guo, H. W.; Hovhannisyan, A.; Ghukasyan, V.; Buryakina, T.; Chen, Y. F.; Dong, C. Y.

2014-01-01

Collagen is the main structural protein and the key determinant of mechanical and functional properties of tissues and organs. Proper balance between synthesis and degradation of collagen molecules is critical for maintaining normal physiological functions. In addition, collagen influences tumor development and drug delivery, which makes it a potential cancer therapy target. Using second harmonic generation, two-photon excited fluorescence microscopy, and spectrofluorimetry, we show that the ...

Multiphoton microscopy: an efficient tool for in-situ study of cultural heritage artifacts

Science.gov (United States)

Latour, Gaël.; Echard, Jean-Philippe; Didier, Marie; Schanne-Klein, Marie-Claire

2013-05-01

We present multimodal nonlinear optical imaging of historical artifacts by combining Two-Photon Excited Fluorescence (2PEF) and Second Harmonic Generation (SHG) microscopies. Three-dimensional (3D) non-contact laser-scanning imaging with micrometer resolution is performed without any preparation of the objects under study. 2PEF signals are emitted by a wide range of fluorophores such as pigments and binder, which can be discriminated thanks to their different emission spectral bands by using suitable spectral filters in the detection channel. SHG signals are specific for dense non-centrosymmetric organizations such as the crystalline cellulose within the wood cell walls. We also show that plaster particles exhibit SHG signals. These particles are bassanite crystals with a non-centrosymmetric crystalline structure, while the other types of calcium sulphates exhibit a centrosymmetric crystalline structure with no SHG signal. In our study, we first characterize model single-layered samples: wood, gelatin-based films containing plaster or cochineal lake and sandarac film containing cochineal lake. We then study multilayered coating systems on wood and show that multimodal nonlinear microscopy successfully reveals the 3D distribution of all components within the stratified sample. We also show that the fine structure of the wood can be assessed, even through a thick multilayered varnish coating. Finally, in situ multimodal nonlinear imaging is demonstrated in a historical violin. SHG/2PEF imaging thus appears as an efficient non-destructive and contactless 3D imaging technique for in situ investigation of historical coatings and more generally for wood characterization and coating analysis at micrometer scale.

Label-free in vivo in situ diagnostic imaging by cellular metabolism quantification with a flexible multiphoton endomicroscope (Conference Presentation)

Science.gov (United States)

Leclerc, Pierre; Hage, Charles-Henri; Fabert, Marc; Brevier, Julien; O'Connor, Rodney P.; Bardet-Coste, Sylvia M.; Habert, Rémi; Braud, Flavie; Kudlinski, Alexandre; Louradour, Frederic

2017-02-01

Multiphoton microscopy is a cutting edge imaging modality leading to increasing advances in biology and also in the clinical field. To use it at its full potential and at the very heart of clinical practice, there have been several developments of fiber-based multiphoton microendoscopes. The application for those probes is now limited by few major restrictions, such as the difficulty to collect autofluorescence signals from tissues and cells theses being inherently weak (e.g. the ones from intracellular NADH or FAD metabolites). This limitation reduces the usefulness of microendoscopy in general, effectively restraining it to morphological imaging modality requiring staining of the tissues. Our aim is to go beyond this limitation, showing for the first time label-free cellular metabolism monitoring, in vivo in situ in real time. The experimental setup is an upgrade of a recently published one (Ducourthial et.al, Scientific Reports, 2016) where femtosecond pulse fiber delivery is further optimized thank's to a new transmissive-GRISM-based pulse stretcher permitting high energy throughput and wide bandwidth. This device allows fast sequential operation with two different excitation wavelengths for efficient two-photon excited NADH and FAD autofluorescence endoscopic detection (i.e. 860 nm for FAD and 760 nm for NADH), enabling cellular optical redox ratio quantification at 8 frames/s. The obtained results on cell models in vitro and also on animal models in vivo (e.g. neurons of a living mouse) prove that we accurately assess the level of NADH and FAD at subcellular resolution through a 3-meters-long fiber with our miniaturized probe (O.D. =2.2 mm).

Safety assessment in primary Mycobacterium tuberculosis smear microscopy centres in Blantyre Malawi: a facility based cross sectional survey.

Science.gov (United States)

Majamanda, J; Ndhlovu, P; Shawa, I T

2013-12-01

Tuberculosis (TB) is caused by Mycobacterium tuberculosis and is transmitted mainly through aerosolization of infected sputum which puts laboratory workers at risk in spite of the laboratory workers' risk of infection being at 3 to 9 times higher than the general public. Laboratory safety should therefore be prioritized and optimized to provide sufficient safety to laboratory workers. To assess the safety for the laboratory workers in TB primary microscopy centres in Blantyre urban. TB primary microscopy centers in Blantyre urban were assessed in aspects of equipment availability, facility layout, and work practice, using a standardized WHO/AFRO ISO 15189 checklist for the developing countries which sets the minimum safety score at ≥80%. Each center was graded according to the score it earned upon assessment. Only one (1) microscopy center out nine (9) reached the minimum safety requirement. Four (4) centers were awarded 1 star level, four (4) centers were awarded 2 star level and only one (1) center was awarded 3 star level. In Blantyre urban, 89% of the Tuberculosis microscopy centers are failing to provide the minimum safety to the laboratory workers. Government and other stake holders should be committed in addressing the safety challenges of TB microscopy centres in the country to ensure safety for the laboratory workers. It is recommended that the study be conducted at the regional or national level for both public and private laboratories in order to have a general picture of safety in Tb microscopy centres possibly across the country.

Temporal shaping of nanosecond CO2 laser pulses in multiphoton saturable absorbers

International Nuclear Information System (INIS)

Haglund, R.F. Jr.

1981-01-01

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

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

Science.gov (United States)

2015-05-21

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

Utilizing nonlinear optical microscopy to investigate the development of early cancer in nude mice in vivo

Science.gov (United States)

Wang, Chun-Chin; Li, Feng-Chieh; Lin, Sung-Jan; Lo, Wen; Dong, Chen-Yuan

2007-07-01

In this investigation, we used in vivo nonlinear optical microscopy to image normal and carcinogen DMBA treated skin tissues of nude mice. We acquired two-photon autofluroescence and second harmonic generation (SHG) images of the skin tissue, and applied the ASI (Autofluorescence versus SHG Index) to the resulting image. This allows us to visualize and quantify the interaction between mouse skin cells and the surrounding connective tissue. We found that as the imaging depth increases, ASI has a different distribution in the normal and the treated skin tissues. Since the DMBA treated skin eventually became squamous cell carcinoma (SCC), our results show that the physiological changes to mouse skin en route to become cancer can be effectively tracked by multiphoton microscopy. We envision this approach to be effective in studying tumor biology and tumor treatment procedures.

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

Science.gov (United States)

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

2018-04-01

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

Monitoring wound healing by multiphoton tomography/endoscopy

Science.gov (United States)

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

2015-02-01

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

Optical Magnetometry Using Multiphoton Transitions

Science.gov (United States)

Degenkolb, Skyler M.

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

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

International Nuclear Information System (INIS)

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

2016-01-01

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

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

Energy Technology Data Exchange (ETDEWEB)

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

2016-09-01

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

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

Science.gov (United States)

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

2017-09-01

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

Clinical studies of pigmented lesions in human skin by using a multiphoton tomograph

Science.gov (United States)

Balu, Mihaela; Kelly, Kristen M.; Zachary, Christopher B.; Harris, Ronald M.; Krasieva, Tatiana B.; König, Karsten; Tromberg, Bruce J.

2013-02-01

In vivo imaging of pigmented lesions in human skin was performed with a clinical multiphoton microscopy (MPM)-based tomograph (MPTflex, JenLab, Germany). Two-photon excited fluorescence was used for visualizing endogenous fluorophores such as NADH/FAD, keratin, melanin in the epidermal cells and elastin fibers in the dermis. Collagen fibers were imaged by second harmonic generation. Our study involved in vivo imaging of benign melanocytic nevi, atypical nevi and melanoma. The goal of this preliminary study was to identify in vivo the characteristic features and their frequency in pigmented lesions at different stages (benign, atypical and malignant) and to evaluate the ability of in vivo MPM to distinguish atypical nevi from melanoma. Comparison with histopathology was performed for the biopsied lesions. Benign melanocytic nevi were characterized by the presence of nevus cell nests at the epidermal-dermal junction. In atypical nevi, features such as lentiginous hyperplasia, acanthosis and architectural disorder were imaged. Cytological atypia was present in all the melanoma lesions imaged, showing the strongest correlation with malignancy. The MPM images demonstrated very good correlation with corresponding histological images, suggesting that MPM could be a promising tool for in vivo non-invasive pigmented lesion diagnosis, particularly distinguishing atypical nevi from melanoma.

Recoil effects in multiphoton electron-positron pair creation

International Nuclear Information System (INIS)

Krajewska, K.; Kaminski, J. Z.

2010-01-01

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

The electron microscopy facility at the LNLS

International Nuclear Information System (INIS)

Ugarte, D.; Zanchet, D.; Silva, P.C.; Araujo, S.R. de; Bettini, J.; Gonzalez, J.C.; Nakabayashi, D.B.

2004-01-01

Full text: The Electron Microscopy Laboratory (LME, Lab. Microscopia Eletronica) is one of the multi user facilities of the Laboratorio Nacional de Luz Sincrotron (LNLS). It has been in operation since the beginning of 1999 to provide spatial high resolution tools, making the LNLS a unique center for advanced characterization of materials. The equipment installed at the LME can be brie y described as: a) a Low Vacuum Scanning Electron Microscope (SEM, JSM-5900LV) with microanalysis and crystallographic mapping capabilities; b) a Field Emission Gun SEM (JSM-6330F); c) a 300 kV High Resolution Transmission Electron Microscope (HRTEM, JEM 3010 URP, 1.7 A Point Res.) with TV Camera, Multi-Scan CCD Camera and X-ray Si(Li) detector; and d) a complete sample preparation laboratory for EM studies A simple procedure allows access to the LME instruments, firstly a short research project must be submitted for evaluation of viability and relevance; subsequently the training microscope sessions are scheduled. It is important to remark that EM is a routine characterization tool and the researchers have to operate the microscope by themselves; for that a training period is necessary, which may vary from 1-2 weeks for a SEM to 2-4 months for the HRTEM. Our staff addresses a great effort to the formation of human resources in order to allow inexperienced Users to become capable of acquiring and interpreting data for their research projects. Since its installation, the LME has trained more than 300 Users in EM techniques. In 2003, the number of projects developed was: 36 in the HRTEM, 16 in the FEG-SEM and 48 in the LV-SEM. This means that just the HRTEM has operated 2157 hours. The constant increase of users in addition to the more exigent EM studies being proposed indicates the necessity of an expansion of the LME by the purchase of a 200 kV FEG-TEM oriented for nano-analysis and Electron Energy Loss Spectroscopy.. (author)

Studies of atmospheric molecules by multiphoton spectroscopy

International Nuclear Information System (INIS)

Johnson, P.M.

1990-12-01

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

Rotational averaging of multiphoton absorption cross sections

Energy Technology Data Exchange (ETDEWEB)

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

2014-11-28

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

Variational methods for high-order multiphoton processes

International Nuclear Information System (INIS)

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

1990-01-01

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

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

Science.gov (United States)

Meyer, Michael

2015-05-01

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

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

Science.gov (United States)

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

2013-05-04

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

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

International Nuclear Information System (INIS)

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

2006-01-01

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

Phenol-ammonium sulfate microscopy method for diagnosis of pulmonary tuberculosis

Directory of Open Access Journals (Sweden)

Ritu Singhal

2013-01-01

Conclusion: Sputum microscopy with PhAS solution is a safe, reliable and inexpensive alternative for direct microscopy. This method can be conveniently applied for usage in microscopy centers with limited bio-safety facilities.

Multiphoton tomography of astronauts

Science.gov (United States)

König, Karsten; Weinigel, Martin; Pietruszka, Anna; Bückle, Rainer; Gerlach, Nicole; Heinrich, Ulrike

2015-03-01

Weightlessness may impair the astronaut's health conditions. Skin impairments belong to the most frequent health problems during space missions. Within the Skin B project, skin physiological changes during long duration space flights are currently investigated on three European astronauts that work for nearly half a year at the ISS. Measurements on the hydration, the transepidermal water loss, the surface structure, elasticity and the tissue density by ultrasound are conducted. Furthermore, high-resolution in vivo histology is performed by multiphoton tomography with 300 nm spatial and 200 ps temporal resolution. The mobile certified medical tomograph with a flexible 360° scan head attached to a mechano-optical arm is employed to measure two-photon autofluorescence and SHG in the volar forearm of the astronauts. Modification of the tissue architecture and of the fluorescent biomolecules NAD(P)H, keratin, melanin and elastin are detected as well as of SHG-active collagen. Thinning of the vital epidermis, a decrease of the autofluoresence intensity, an increase in the long fluorescence lifetime, and a reduced skin ageing index SAAID based on an increased collagen level in the upper dermis have been found. Current studies focus on recovery effects.

Photo-induced processes in collagen-hypericin system revealed by fluorescence spectroscopy and multiphoton microscopy.

Science.gov (United States)

Hovhannisyan, V; Guo, H W; Hovhannisyan, A; Ghukasyan, V; Buryakina, T; Chen, Y F; Dong, C Y

2014-05-01

Collagen is the main structural protein and the key determinant of mechanical and functional properties of tissues and organs. Proper balance between synthesis and degradation of collagen molecules is critical for maintaining normal physiological functions. In addition, collagen influences tumor development and drug delivery, which makes it a potential cancer therapy target. Using second harmonic generation, two-photon excited fluorescence microscopy, and spectrofluorimetry, we show that the natural pigment hypericin induces photosensitized destruction of collagen-based tissues. We demonstrate that hypericin-mediated processes in collagen fibers are irreversible and may be used for the treatment of cancer and collagen-related disorders.

Second harmonic generation microscopy differentiates collagen type I and type III in COPD

Science.gov (United States)

Suzuki, Masaru; Kayra, Damian; Elliott, W. Mark; Hogg, James C.; Abraham, Thomas

2012-03-01

The structural remodeling of extracellular matrix proteins in peripheral lung region is an important feature in chronic obstructive pulmonary disease (COPD). Multiphoton microscopy is capable of inducing specific second harmonic generation (SHG) signal from non-centrosymmetric structural proteins such as fibrillar collagens. In this study, SHG microscopy was used to examine structural remodeling of the fibrillar collagens in human lungs undergoing emphysematous destruction (n=2). The SHG signals originating from these diseased lung thin sections from base to apex (n=16) were captured simultaneously in both forward and backward directions. We found that the SHG images detected in the forward direction showed well-developed and well-structured thick collagen fibers while the SHG images detected in the backward direction showed striking different morphological features which included the diffused pattern of forward detected structures plus other forms of collagen structures. Comparison of these images with the wellestablished immunohistochemical staining indicated that the structures detected in the forward direction are primarily the thick collagen type I fibers and the structures identified in the backward direction are diffusive structures of forward detected collagen type I plus collagen type III. In conclusion, we here demonstrate the feasibility of SHG microscopy in differentiating fibrillar collagen subtypes and understanding their remodeling in diseased lung tissues.

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

Science.gov (United States)

Bell, Alex; Kofron, Matthew; Nistor, Vasile

2015-09-03

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

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

Science.gov (United States)

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

2018-04-01

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

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

DEFF Research Database (Denmark)

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

2017-01-01

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

Examination of excited state populations in sputtering using Multiphoton Resonance Ionization

International Nuclear Information System (INIS)

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

1984-01-01

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

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

Indian Academy of Sciences (India)

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

Ionisation of hydrogen-like atoms by a multiphoton absorption process

International Nuclear Information System (INIS)

Gontier, Y.; Trahin, M.

1967-01-01

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

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

NARCIS (Netherlands)

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

1997-01-01

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

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

Energy Technology Data Exchange (ETDEWEB)

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

2013-12-21

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

Infrared multiphoton absorption and decomposition

International Nuclear Information System (INIS)

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

1984-01-01

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

Picosecond multiphoton ionization of atomic and molecular clusters

International Nuclear Information System (INIS)

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

1990-01-01

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

Clinical multiphoton tomography and clinical two-photon microendoscopy

Science.gov (United States)

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

2009-02-01

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

Optimizing pulse compressibility in completely all-fibered Ytterbium chirped pulse amplifiers for in vivo two photon laser scanning microscopy.

Science.gov (United States)

Fernández, A; Grüner-Nielsen, L; Andreana, M; Stadler, M; Kirchberger, S; Sturtzel, C; Distel, M; Zhu, L; Kautek, W; Leitgeb, R; Baltuska, A; Jespersen, K; Verhoef, A

2017-08-01

A simple and completely all-fiber Yb chirped pulse amplifier that uses a dispersion matched fiber stretcher and a spliced-on hollow core photonic bandgap fiber compressor is applied in nonlinear optical microscopy. This stretching-compression approach improves compressibility and helps to maximize the fluorescence signal in two-photon laser scanning microscopy as compared with approaches that use standard single mode fibers as stretcher. We also show that in femtosecond all-fiber systems, compensation of higher order dispersion terms is relevant even for pulses with relatively narrow bandwidths for applications relying on nonlinear optical effects. The completely all-fiber system was applied to image green fluorescent beads, a stained lily-of-the-valley root and rat-tail tendon. We also demonstrated in vivo imaging in zebrafish larvae, where we simultaneously measure second harmonic and fluorescence from two-photon excited red-fluorescent protein. Since the pulses are compressed in a fiber, this source is especially suited for upgrading existing laser scanning (confocal) microscopes with multiphoton imaging capabilities in space restricted settings or for incorporation in endoscope-based microscopy.

Examination of excited state populations in sputtering using multiphoton resonance ionization

International Nuclear Information System (INIS)

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

1984-01-01

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

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

International Nuclear Information System (INIS)

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

1994-01-01

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

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

DEFF Research Database (Denmark)

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

2013-01-01

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

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

Energy Technology Data Exchange (ETDEWEB)

Hu, Huayu

2011-04-27

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

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

International Nuclear Information System (INIS)

Hu, Huayu

2011-01-01

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

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

CERN Document Server

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

2004-01-01

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

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

International Nuclear Information System (INIS)

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

1989-01-01

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

The study of multiphoton ionization processes in hydrogen atoms

International Nuclear Information System (INIS)

Mohammad, M.A.

1981-01-01

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

A phasor approach analysis of multiphoton FLIM measurements of three-dimensional cell culture models

Science.gov (United States)

Lakner, P. H.; Möller, Y.; Olayioye, M. A.; Brucker, S. Y.; Schenke-Layland, K.; Monaghan, M. G.

2016-03-01

Fluorescence lifetime imaging microscopy (FLIM) is a useful approach to obtain information regarding the endogenous fluorophores present in biological samples. The concise evaluation of FLIM data requires the use of robust mathematical algorithms. In this study, we developed a user-friendly phasor approach for analyzing FLIM data and applied this method on three-dimensional (3D) Caco-2 models of polarized epithelial luminal cysts in a supporting extracellular matrix environment. These Caco-2 based models were treated with epidermal growth factor (EGF), to stimulate proliferation in order to determine if FLIM could detect such a change in cell behavior. Autofluorescence from nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) in luminal Caco-2 cysts was stimulated by 2-photon laser excitation. Using a phasor approach, the lifetimes of involved fluorophores and their contribution were calculated with fewer initial assumptions when compared to multiexponential decay fitting. The phasor approach simplified FLIM data analysis, making it an interesting tool for non-experts in numerical data analysis. We observed that an increased proliferation stimulated by EGF led to a significant shift in fluorescence lifetime and a significant alteration of the phasor data shape. Our data demonstrates that multiphoton FLIM analysis with the phasor approach is a suitable method for the non-invasive analysis of 3D in vitro cell culture models qualifying this method for monitoring basic cellular features and the effect of external factors.

Multiphoton amplitude in a constant background field

Science.gov (United States)

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

2018-01-01

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

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

International Nuclear Information System (INIS)

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

2005-01-01

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

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

Energy Technology Data Exchange (ETDEWEB)

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

2013-08-07

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

Multiphoton dissociation and thermal unimolecular reactions induced by infrared lasers

International Nuclear Information System (INIS)

Dai, H.L.

1981-04-01

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

An enhancement of spin polarization by multiphoton pumping in semiconductors

Energy Technology Data Exchange (ETDEWEB)

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

2011-08-15

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

An enhancement of spin polarization by multiphoton pumping in semiconductors

International Nuclear Information System (INIS)

Miah, M. Idrish

2011-01-01

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

Multiphoton ionization of atomic cesium

International Nuclear Information System (INIS)

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

1984-01-01

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

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

International Nuclear Information System (INIS)

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

1987-01-01

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

In vivo non-invasive monitoring of collagen remodelling by two-photon microscopy after micro-ablative fractional laser resurfacing.

Science.gov (United States)

Cicchi, Riccardo; Kapsokalyvas, Dimitrios; Troiano, Michela; Campolmi, Piero; Morini, Cristiano; Massi, Daniela; Cannarozzo, Giovanni; Lotti, Torello; Pavone, Francesco Saverio

2014-11-01

Non-linear optical microscopy is becoming popular as a non-invasive in vivo imaging modality in dermatology. In this study, combined TPF and SHG microscopy were used to monitor collagen remodelling in vivo after micro-ablative fractional laser resurfacing. Papillary dermis of living subjects, covering a wide age range, was imaged immediately before and forty days after treatment. A qualitative visual examination of acquired images demonstrated an age-dependent remodelling effect on collagen. Additional quantitative analysis of new collagen production was performed by means of two image analysis methods. A higher increase in SHG to TPF ratio, corresponding to a stronger treatment effectiveness, was found in older subjects, whereas the effect was found to be negligible in young, and minimal in middle age subjects. Analysis of collagen images also showed a dependence of the treatment effectiveness with age but with controversial results. While the diagnostic potential of in vivo multiphoton microscopy has already been demonstrated for skin cancer and other skin diseases, here we first successfully explore its potential use for a non-invasive follow-up of a laser-based treatment. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Search for anomalous multiphoton production at 100-300 GeV

International Nuclear Information System (INIS)

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

1975-01-01

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

3D Printer Generated Tissue iMolds for Cleared Tissue Using Single- and Multi-Photon Microscopy for Deep Tissue Evaluation.

Science.gov (United States)

Miller, Sean J; Rothstein, Jeffrey D

2017-01-01

Pathological analyses and methodology has recently undergone a dramatic revolution. With the creation of tissue clearing methods such as CLARITY and CUBIC, groups can now achieve complete transparency in tissue samples in nano-porous hydrogels. Cleared tissue is then imagined in a semi-aqueous medium that matches the refractive index of the objective being used. However, one major challenge is the ability to control tissue movement during imaging and to relocate precise locations post sequential clearing and re-staining. Using 3D printers, we designed tissue molds that fit precisely around the specimen being imaged. First, images are taken of the specimen, followed by importing and design of a structural mold, then printed with affordable plastics by a 3D printer. With our novel design, we have innovated tissue molds called innovative molds (iMolds) that can be generated in any laboratory and are customized for any organ, tissue, or bone matter being imaged. Furthermore, the inexpensive and reusable tissue molds are made compatible for any microscope such as single and multi-photon confocal with varying stage dimensions. Excitingly, iMolds can also be generated to hold multiple organs in one mold, making reconstruction and imaging much easier. Taken together, with iMolds it is now possible to image cleared tissue in clearing medium while limiting movement and being able to relocate precise anatomical and cellular locations on sequential imaging events in any basic laboratory. This system provides great potential for screening widespread effects of therapeutics and disease across entire organ systems.

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

International Nuclear Information System (INIS)

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

1985-01-01

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

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

Science.gov (United States)

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

1988-10-01

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

Proximal Probes Facility

Data.gov (United States)

Federal Laboratory Consortium — The Proximal Probes Facility consists of laboratories for microscopy, spectroscopy, and probing of nanostructured materials and their functional properties. At the...

Multiphoton autofluorescence lifetime imaging of induced pluripotent stem cells

Science.gov (United States)

Uchugonova, Aisada

2017-06-01

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

Multiphoton transitions in semiconductors in the non-perturbative approach

International Nuclear Information System (INIS)

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

1987-09-01

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

Advanced multiphoton methods for in vitro and in vivo functional imaging of mouse retinal neurons (Conference Presentation)

Science.gov (United States)

Cohen, Noam; Schejter, Adi; Farah, Nairouz; Shoham, Shy

2016-03-01

Studying the responses of retinal ganglion cell (RGC) populations has major significance in vision research. Multiphoton imaging of optogenetic probes has recently become the leading approach for visualizing neural populations and has specific advantages for imaging retinal activity during visual stimulation, because it leads to reduced direct photoreceptor excitation. However, multiphoton retinal activity imaging is not straightforward: point-by-point scanning leads to repeated neural excitation while optical access through the rodent eye in vivo has proven highly challenging. Here, we present two enabling optical designs for multiphoton imaging of responses to visual stimuli in mouse retinas expressing calcium indicators. First, we present an imaging solution based on Scanning Line Temporal Focusing (SLITE) for rapidly imaging neuronal activity in vitro. In this design, we scan a temporally focused line rather than a point, increasing the scan speed and reducing the impact of repeated excitation, while maintaining high optical sectioning. Second, we present the first in vivo demonstration of two-photon imaging of RGC activity in the mouse retina. To obtain these cellular resolution recordings we integrated an illumination path into a correction-free imaging system designed using an optical model of the mouse eye. This system can image at multiple depths using an electronically tunable lens integrated into its optical path. The new optical designs presented here overcome a number of outstanding obstacles, allowing the study of rapid calcium- and potentially even voltage-indicator signals both in vitro and in vivo, thereby bringing us a step closer toward distributed monitoring of action potentials.

Second harmonic generation microscopy of the living human cornea

Science.gov (United States)

Artal, Pablo; Ávila, Francisco; Bueno, Juan

2018-02-01

Second Harmonic Generation (SHG) microscopy provides high-resolution structural imaging of the corneal stroma without the need of labelling techniques. This powerful tool has never been applied to living human eyes so far. Here, we present a new compact SHG microscope specifically developed to image the structural organization of the corneal lamellae in living healthy human volunteers. The research prototype incorporates a long-working distance dry objective that allows non-contact three-dimensional SHG imaging of the cornea. Safety assessment and effectiveness of the system were firstly tested in ex-vivo fresh eyes. The maximum average power of the used illumination laser was 20 mW, more than 10 times below the maximum permissible exposure (according to ANSI Z136.1-2000). The instrument was successfully employed to obtain non-contact and non-invasive SHG of the living human eye within well-established light safety limits. This represents the first recording of in vivo SHG images of the human cornea using a compact multiphoton microscope. This might become an important tool in Ophthalmology for early diagnosis and tracking ocular pathologies.

Nonlinear adaptive optics: aberration correction in three photon fluorescence microscopy for mouse brain imaging

Science.gov (United States)

Sinefeld, David; Paudel, Hari P.; Wang, Tianyu; Wang, Mengran; Ouzounov, Dimitre G.; Bifano, Thomas G.; Xu, Chris

2017-02-01

Multiphoton fluorescence microscopy is a well-established technique for deep-tissue imaging with subcellular resolution. Three-photon microscopy (3PM) when combined with long wavelength excitation was shown to allow deeper imaging than two-photon microscopy (2PM) in biological tissues, such as mouse brain, because out-of-focus background light can be further reduced due to the higher order nonlinear excitation. As was demonstrated in 2PM systems, imaging depth and resolution can be improved by aberration correction using adaptive optics (AO) techniques which are based on shaping the scanning beam using a spatial light modulator (SLM). In this way, it is possible to compensate for tissue low order aberration and to some extent, to compensate for tissue scattering. Here, we present a 3PM AO microscopy system for brain imaging. Soliton self-frequency shift is used to create a femtosecond source at 1675 nm and a microelectromechanical (MEMS) SLM serves as the wavefront shaping device. We perturb the 1020 segment SLM using a modified nonlinear version of three-point phase shifting interferometry. The nonlinearity of the fluorescence signal used for feedback ensures that the signal is increasing when the spot size decreases, allowing compensation of phase errors in an iterative optimization process without direct phase measurement. We compare the performance for different orders of nonlinear feedback, showing an exponential growth in signal improvement as the nonlinear order increases. We demonstrate the impact of the method by applying the 3PM AO system for in-vivo mouse brain imaging, showing improvement in signal at 1-mm depth inside the brain.

Experimental quantum teleportation and multiphoton entanglement via interfering narrowband photon sources

International Nuclear Information System (INIS)

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

2009-01-01

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

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

Energy Technology Data Exchange (ETDEWEB)

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

2010-08-23

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

Multi-photon vertical cross-sectional imaging with a dynamically-balanced thin-film PZT z-axis microactuator.

Science.gov (United States)

Choi, Jongsoo; Duan, Xiyu; Li, Haijun; Wang, Thomas D; Oldham, Kenn R

2017-10-01

Use of a thin-film piezoelectric microactuator for axial scanning during multi-photon vertical cross-sectional imaging is described. The actuator uses thin-film lead-zirconate-titanate (PZT) to generate upward displacement of a central mirror platform, micro-machined from a silicon-on-insulator (SOI) wafer to dimensions compatible with endoscopic imaging instruments. Device modeling in this paper focuses on existence of frequencies near device resonance producing vertical motion with minimal off-axis tilt even in the presence of multiple vibration modes and non-uniformity in fabrication outcomes. Operation near rear resonance permits large stroke lengths at low voltages relative to other vertical microactuators. Highly uniform vertical motion of the mirror platform is a key requirement for vertical cross-sectional imaging in the remote scan architecture being used for multi-photon instrument prototyping. The stage is installed in a benchtop testbed in combination with an electrostatic mirror that performs in-plane scanning. Vertical sectional images are acquired from 15 μm diameter beads and excised mouse colon tissue.

Multiphoton Absorption Order of CsPbBr3 As Determined by Wavelength-Dependent Nonlinear Optical Spectroscopy.

Science.gov (United States)

Saouma, Felix O; Stoumpos, Constantinos C; Kanatzidis, Mercouri G; Kim, Yong Soo; Jang, Joon I

2017-10-05

CsPbBr 3 is a direct-gap semiconductor where optical absorption takes place across the fundamental bandgap, but this all-inorganic halide perovskite typically exhibits above-bandgap emission when excited over an energy level, lying above the conduction-band minimum. We probe this bandgap anomaly using wavelength-dependent multiphoton absorption spectroscopy and find that the fundamental gap is strictly two-photon forbidden, rendering it three-photon absorption (3PA) active. Instead, two-photon absorption (2PA) commences when the two-photon energy is resonant with the optical gap, associated with the level causing the anomaly. We determine absolute nonlinear optical dispersion over this 3PA-2PA region, which can be explained by two-band models in terms of the optical gap. The polarization dependence of 3PA and 2PA is also measured and explained by the relevant selection rules. CsPbBr 3 is highly luminescent under multiphoton absorption at room temperature with marked polarization and wavelength dependence at the 3PA-2PA crossover and therefore has potential for nonlinear optical applications.

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

International Nuclear Information System (INIS)

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

2005-01-01

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

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

DEFF Research Database (Denmark)

Mangiarotti, Alessio; Sona, Pietro; Ballestrero, Sergio

2012-01-01

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

Scanning photoemission microscopy with synchrotron radiation

Science.gov (United States)

Ade, Harald W.

1992-08-01

Progress in photoemission spectro-microscopy at various synchrotron radiation facilities is reviewed. Microprobe devices such as MAXIMUM at the SRC in Wisconsin, the X1-SPEM at the NSLS at BNL, as well as the ellipsoidal ring mirror microscope at DESY in Hamburg, recorded first images during the last few years. The present status of these devices which achieve their lateral resolution by focusing X-rays to a small spot is the primary focus of this paper, but work representing other approaches to spectro-microscopy is also discussed.

Multiphoton ionization of atomic cesium

International Nuclear Information System (INIS)

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

1984-01-01

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

Correlative two-photon and serial block face scanning electron microscopy in neuronal tissue using 3D near-infrared branding maps.

Science.gov (United States)

Lees, Robert M; Peddie, Christopher J; Collinson, Lucy M; Ashby, Michael C; Verkade, Paul

2017-01-01

Linking cellular structure and function has always been a key goal of microscopy, but obtaining high resolution spatial and temporal information from the same specimen is a fundamental challenge. Two-photon (2P) microscopy allows imaging deep inside intact tissue, bringing great insight into the structural and functional dynamics of cells in their physiological environment. At the nanoscale, the complex ultrastructure of a cell's environment in tissue can be reconstructed in three dimensions (3D) using serial block face scanning electron microscopy (SBF-SEM). This provides a snapshot of high resolution structural information pertaining to the shape, organization, and localization of multiple subcellular structures at the same time. The pairing of these two imaging modalities in the same specimen provides key information to relate cellular dynamics to the ultrastructural environment. Until recently, approaches to relocate a region of interest (ROI) in tissue from 2P microscopy for SBF-SEM have been inefficient or unreliable. However, near-infrared branding (NIRB) overcomes this by using the laser from a multiphoton microscope to create fiducial markers for accurate correlation of 2P and electron microscopy (EM) imaging volumes. The process is quick and can be user defined for each sample. Here, to increase the efficiency of ROI relocation, multiple NIRB marks are used in 3D to target ultramicrotomy. A workflow is described and discussed to obtain a data set for 3D correlated light and electron microscopy, using three different preparations of brain tissue as examples. Copyright © 2017 Elsevier Inc. All rights reserved.

Probing stem cell differentiation using atomic force microscopy

International Nuclear Information System (INIS)

Liang, Xiaobin; Shi, Xuetao; Ostrovidov, Serge; Wu, Hongkai; Nakajima, Ken

2016-01-01

Graphical abstract: - Highlights: • Atomic force microscopy (AFM) was developed to probe stem cell differentiation. • The mechanical properties of stem cells and their ECMs can be used to clearly distinguish specific stem cell-differentiated lineages. • AFM is a facile and useful tool for monitoring stem cell differentiation in a non-invasive manner. - Abstract: A real-time method using atomic force microscopy (AFM) was developed to probe stem cell differentiation by measuring the mechanical properties of cells and the extracellular matrix (ECM). The mechanical properties of stem cells and their ECMs can be used to clearly distinguish specific stem cell-differentiated lineages. It is clear that AFM is a facile and useful tool for monitoring the differentiation of stem cells in a non-invasive manner.

Probing stem cell differentiation using atomic force microscopy

Energy Technology Data Exchange (ETDEWEB)

Liang, Xiaobin [Graduate School of Science and Engineering, Tokyo Institute of Technology, Ookayama 2-12-1, Meguro-ku, Tokyo 152-8550 (Japan); Shi, Xuetao, E-mail: mrshixuetao@gmail.com [School of Materials Science and Engineering, South China University of Technology, Guangzhou 510641 (China); Ostrovidov, Serge [WPI-Advanced Institute for Materials Research, Tohoku University, Sendai (Japan); Wu, Hongkai, E-mail: chhkwu@ust.hk [Department of Chemistry & Division of Biomedical Engineering, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong (China); Nakajima, Ken [Graduate School of Science and Engineering, Tokyo Institute of Technology, Ookayama 2-12-1, Meguro-ku, Tokyo 152-8550 (Japan)

2016-03-15

Graphical abstract: - Highlights: • Atomic force microscopy (AFM) was developed to probe stem cell differentiation. • The mechanical properties of stem cells and their ECMs can be used to clearly distinguish specific stem cell-differentiated lineages. • AFM is a facile and useful tool for monitoring stem cell differentiation in a non-invasive manner. - Abstract: A real-time method using atomic force microscopy (AFM) was developed to probe stem cell differentiation by measuring the mechanical properties of cells and the extracellular matrix (ECM). The mechanical properties of stem cells and their ECMs can be used to clearly distinguish specific stem cell-differentiated lineages. It is clear that AFM is a facile and useful tool for monitoring the differentiation of stem cells in a non-invasive manner.

Development of resonance-enhanced multiphoton ionization system

International Nuclear Information System (INIS)

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

2009-05-01

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

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

Science.gov (United States)

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

1995-07-01

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

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

International Nuclear Information System (INIS)

Requate, A.

2007-03-01

Theoretical analysis of the vibrational excitation of small molecules during multiphoton ionization in intense laser fields of optical and infrared frequencies. Analysis of the alignment dependence of the electron impact ionization of diatomic molecules in the presence of an intense laser field as the final step in the process of Nonsequential Double Ionization. Quantum mechanical description using S-matrix theory in Strong Field Approximation (SFA), i.e. beyond perturbation theory. (orig.)

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

Energy Technology Data Exchange (ETDEWEB)

Requate, A

2007-03-15

Theoretical analysis of the vibrational excitation of small molecules during multiphoton ionization in intense laser fields of optical and infrared frequencies. Analysis of the alignment dependence of the electron impact ionization of diatomic molecules in the presence of an intense laser field as the final step in the process of Nonsequential Double Ionization. Quantum mechanical description using S-matrix theory in Strong Field Approximation (SFA), i.e. beyond perturbation theory. (orig.)

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

Czech Academy of Sciences Publication Activity Database

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

2016-01-01

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

Multiphoton Rabi oscillations between highly excited Stark states of potassium

International Nuclear Information System (INIS)

He Yonglin

2011-01-01

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

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

International Nuclear Information System (INIS)

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

1989-01-01

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

Search for Multiphoton Signatures of a Higgs Boson

Energy Technology Data Exchange (ETDEWEB)

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

2006-07-01

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

Light Microscopy Module (LMM)-Emulator

Science.gov (United States)

Levine, Howard G.; Smith, Trent M.; Richards, Stephanie E.

2016-01-01

The Light Microscopy Module (LMM) is a microscope facility developed at Glenn Research Center (GRC) that provides researchers with powerful imaging capability onboard the International Space Station (ISS). LMM has the ability to have its hardware recongured on-orbit to accommodate a wide variety of investigations, with the capability of remotely acquiring and downloading digital images across multiple levels of magnication.

The multiphoton ionization of uranium hexafluoride

International Nuclear Information System (INIS)

Armstrong, D.P.

1992-05-01

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

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

Science.gov (United States)

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

1997-11-01

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

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

Science.gov (United States)

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

2012-11-01

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

Multiphoton spectral analysis of benzo[a]pyrene uptake and metabolism in a rat liver cell line

International Nuclear Information System (INIS)

Barhoumi, Rola; Mouneimne, Youssef; Ramos, Ernesto; Morisseau, Christophe; Hammock, Bruce D.; Safe, Stephen; Parrish, Alan R.; Burghardt, Robert C.

2011-01-01

Dynamic analysis of the uptake and metabolism of polycyclic aromatic hydrocarbons (PAHs) and their metabolites within live cells in real time has the potential to provide novel insights into genotoxic and non-genotoxic mechanisms of cellular injury caused by PAHs. The present work, combining the use of metabolite spectra generated from metabolite standards using multiphoton spectral analysis and an 'advanced unmixing process', identifies and quantifies the uptake, partitioning, and metabolite formation of one of the most important PAHs (benzo[a]pyrene, BaP) in viable cultured rat liver cells over a period of 24 h. The application of the advanced unmixing process resulted in the simultaneous identification of 8 metabolites in live cells at any single time. The accuracy of this unmixing process was verified using specific microsomal epoxide hydrolase inhibitors, glucuronidation and sulfation inhibitors as well as several mixtures of metabolite standards. Our findings prove that the two-photon microscopy imaging surpasses the conventional fluorescence imaging techniques and the unmixing process is a mathematical technique that seems applicable to the analysis of BaP metabolites in living cells especially for analysis of changes of the ultimate carcinogen benzo[a]pyrene-r-7,t-8-dihydrodiol-t-9,10-epoxide. Therefore, the combination of the two-photon acquisition with the unmixing process should provide important insights into the cellular and molecular mechanisms by which BaP and other PAHs alter cellular homeostasis.

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

Energy Technology Data Exchange (ETDEWEB)

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

2014-10-14

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

Development of fiber lasers and devices for coherent Raman scattering microscopy

Science.gov (United States)

Lamb, Erin Stranford

As ultrafast laser technology has found expanding application in machining, spectroscopy, microscopy, surgery, and numerous other areas, the desire for inexpensive and robust laser sources has grown. Until recently, nonlinear effects in fiber systems due to the tight confinement of the light in the core have limited their performance. However, with advances in managing nonlinearity through pulse propagation physics and the use of large core fibers, the performance of fiber lasers can compete with that of their solid-state counterparts. As specific applications, such as coherent Raman scattering microscopy, emerge that stand to benefit from fiber technology, new performance challenges in areas such as laser noise are anticipated. This thesis studies nonlinear pulse propagation in fiber lasers and fiber parametric devices. Applications of dissipative solitons and self-similar pulse propagation to low-repetition rate oscillators that have the potential to simplify short-pulse amplification schemes will be examined. The rest of this thesis focuses on topics relevant to fiber laser development for coherent Raman scattering microscopy sources. Coherent pulse division and recombination inside the laser cavity will be introduced as an energy-scaling mechanism and demonstrated for a fiber soliton laser. The relative intensity noise properties of mode-locked fiber lasers, with a particular emphasis on normal dispersion lasers, will be explored in simulation and experiment. A fiber optical parametric oscillator will be studied in detail for low noise frequency conversion of picosecond pulses, and its utility for coherent Raman imaging will be demonstrated. Spectral compression of femtosecond pulses is used to generate picosecond pulses to pump this device, and this technique provides a route to future noise reduction in the system. Furthermore, this device forms a multimodal source capable of providing the picosecond pulses for coherent Raman scattering microscopy and the

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.

Determination of the Goos-Hänchen shift in dielectric waveguides via photo emission electron microscopy in the visible spectrum.

Science.gov (United States)

Stenmark, Theodore; Word, R C; Könenkamp, R

2016-02-22

Photoemission Electron Microscopy (PEEM) is a versatile tool that relies on the photoelectric effect to produce high-resolution images. Pulse lasers allow for multi-photon PEEM where multiple photons are required excite a single electron. This non-linear process can directly image the near field region of electromagnetic fields in materials. We use this ability here to analyze wave propagation in a linear dielectric waveguide with wavelengths of 410 nm and 780 nm. The propagation constant of the waveguide can be extracted from the interference pattern created by the coupled and incident light and shows distinct polarization dependence. The electromagnetic field interaction at the boundaries can then be deduced which is essential to understand power flow in wave guiding structures. These results match well with simulations using finite element techniques.

Guide to user facilities at the Lawrence Berkeley Laboratory

International Nuclear Information System (INIS)

1984-04-01

Lawrence Berkeley Laboratories' user facilities are described. Specific facilities include: the National Center for Electron Microscopy; the Bevalac; the SuperHILAC; the Neutral Beam Engineering Test Facility; the National Tritium Labeling Facility; the 88 inch Cyclotron; the Heavy Charged-Particle Treatment Facility; the 2.5 MeV Van de Graaff; the Sky Simulator; the Center for Computational Seismology; and the Low Background Counting Facility

Ultrafast Science Opportunities with Electron Microscopy

Energy Technology Data Exchange (ETDEWEB)

DURR, HERMANN; Wang, X.J., ed.

2016-04-28

X-rays and electrons are two of the most fundamental probes of matter. When the Linac Coherent Light Source (LCLS), the world’s first x-ray free electron laser, began operation in 2009, it transformed ultrafast science with the ability to generate laser-like x-ray pulses from the manipulation of relativistic electron beams. This document describes a similar future transformation. In Transmission Electron Microscopy, ultrafast relativistic (MeV energy) electron pulses can achieve unsurpassed spatial and temporal resolution. Ultrafast temporal resolution will be the next frontier in electron microscopy and can ideally complement ultrafast x-ray science done with free electron lasers. This document describes the Grand Challenge science opportunities in chemistry, material science, physics and biology that arise from an MeV ultrafast electron diffraction & microscopy facility, especially when coupled with linac-based intense THz and X-ray pump capabilities.

Multimodal microscopy of immune cells and melanoma for longitudinal studies

Science.gov (United States)

Entenberg, David; Aranda, Iana; Li, Yongbiao; Toledo-Crow, Ricardo; Schaer, David; Li, Yanyun

2006-02-01

Intravital microscopy of cancer is a well established tool that provides direct visualization of the tumor cycle. It traditionally involves one of several strategies: invasive subcutaneous (SC) implantation of tumors followed by surgical opening of skin flaps for imaging, techniques utilizing skin fold chambers and implanted optical windows or intradermal injections under 200μm from the skin surface. All of these techniques allow the use of fluorescent proteins as markers for biologically significant constituents. However, observation methods utilizing skin-flaps, skin-fold chambers and optical windows are invasive and tend to alter the immune environment of the tissue and/or limit the duration of studies that can be performed. If implanted correctly, intradermally injected tumors can be minimally invasive, will not require biopsies or surgical intervention to observe and are accessible for direct transdermal imaging with a number of in vivo modalities. We present our work in the development of a small animal intravital microscopy workstation that allows the acquisition of different contrast imaging modalities: reflectance confocal, wide field epifluorescence, multiphoton and second harmonic generation (SHG). The images are acquired pair-wise simultaneously and sequentially in time. The aim of our instrumentation is to gather all information generated by the single probing beam via the reflected or back-scattered signal, SHG signal and various fluorescence signals. Additionally, we also present our development of a microscopic tissue navigation technique to mark, label and track sites of interest. This technique enables us to revisit sites periodically and record, with different imaging contrasts, their biological changes over time.

Label-free 3D visualization of cellular and tissue structures in intact muscle with second and third harmonic generation microscopy.

Directory of Open Access Journals (Sweden)

Markus Rehberg

Full Text Available Second and Third Harmonic Generation (SHG and THG microscopy is based on optical effects which are induced by specific inherent physical properties of a specimen. As a multi-photon laser scanning approach which is not based on fluorescence it combines the advantages of a label-free technique with restriction of signal generation to the focal plane, thus allowing high resolution 3D reconstruction of image volumes without out-of-focus background several hundred micrometers deep into the tissue. While in mammalian soft tissues SHG is mostly restricted to collagen fibers and striated muscle myosin, THG is induced at a large variety of structures, since it is generated at interfaces such as refraction index changes within the focal volume of the excitation laser. Besides, colorants such as hemoglobin can cause resonance enhancement, leading to intense THG signals. We applied SHG and THG microscopy to murine (Mus musculus muscles, an established model system for physiological research, to investigate their potential for label-free tissue imaging. In addition to collagen fibers and muscle fiber substructure, THG allowed us to visualize blood vessel walls and erythrocytes as well as white blood cells adhering to vessel walls, residing in or moving through the extravascular tissue. Moreover peripheral nerve fibers could be clearly identified. Structure down to the nuclear chromatin distribution was visualized in 3D and with more detail than obtainable by bright field microscopy. To our knowledge, most of these objects have not been visualized previously by THG or any label-free 3D approach. THG allows label-free microscopy with inherent optical sectioning and therefore may offer similar improvements compared to bright field microscopy as does confocal laser scanning microscopy compared to conventional fluorescence microscopy.

Laboratory design for high-performance electron microscopy

Energy Technology Data Exchange (ETDEWEB)

O' Keefe, Michael A.; Turner, John H.; Hetherington, Crispin J.D.; Cullis, A.G.; Carragher, Bridget; Jenkins, Ron; Milgrim, Julie; Milligan,Ronald A.; Potter, Clinton S.; Allard, Lawrence F.; Blom, Douglas A.; Degenhardt, Lynn; Sides, William H.

2004-04-23

Proliferation of electron microscopes with field emission guns, imaging filters and hardware spherical aberration correctors (giving higher spatial and energy resolution) has resulted in the need to construct special laboratories. As resolutions improve, transmission electron microscopes (TEMs) and scanning transmission electron microscopes (STEMs) become more sensitive to ambient conditions. State-of-the-art electron microscopes require state-of-the-art environments, and this means careful design and implementation of microscope sites, from the microscope room to the building that surrounds it. Laboratories have been constructed to house high-sensitive instruments with resolutions ranging down to sub-Angstrom levels; we present the various design philosophies used for some of these laboratories and our experiences with them. Four facilities are described: the National Center for Electron Microscopy OAM Laboratory at LBNL; the FEGTEM Facility at the University of Sheffield; the Center for Integrative Molecular Biosciences at TSRI; and the Advanced Microscopy Laboratory at ORNL.

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

International Nuclear Information System (INIS)

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

2014-01-01

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

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

Energy Technology Data Exchange (ETDEWEB)

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

2014-10-24

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

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

Science.gov (United States)

2009-09-01

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

The New Electron Microscopy: Cells and Molecules in Three Dimensions | Poster

Science.gov (United States)

NCI recently announced the launch of the new National Cryo-Electron Microscopy Facility (NCEF) at the Frederick National Laboratory for Cancer Research (FNLCR). The launch comes while cryo-electron microscopy (cryo-EM) is enjoying the spotlight as a newly emerging, rapidly evolving technology with the potential to revolutionize the field of structural biology. Read more...

Facile synthesis and electron transport properties of NiO nanostructures investigated by scanning tunneling microscopy

Directory of Open Access Journals (Sweden)

Govind Mallick

2017-08-01

Full Text Available Due to their unique chemical, thermal, electronic and photonic properties, low -dimensional transition metal oxides, especially NiO, have attracted great deal of attention for potential applications in a wide range of technologies, such as, sensors, electrochromic coatings and self-healing materials. However, their synthesis involves multi-step complex procedures that in addition to being expensive, further introduce impurities. Here we present a low cost facile approach to synthesize uniform size NiO nanoparticles (NPs from hydrothermally grown Ni(OH2. Detailed transmission electron microscopic analysis reveal the average size of NiO NPs to be around 29 nm. The dimension of NiO NP is also corroborated by the small area scanning tunneling microscope (STM measurements. Further, we investigate electron transport characteristics of newly synthesized Ni(OH2 and NiO nanoparticles on p-type Si substrate using scanning tunneling microscopy. The conductivity of Ni(OH2 and NiO are determined to be 1.46x10-3 S/cm and 2.37x10-5 S/cm, respectively. The NiO NPs exhibit a lower voltage window (∼0.7 V electron tunneling than the parent Ni(OH2.

Facile method to stain the bacterial cell surface for super-resolution fluorescence microscopy

Energy Technology Data Exchange (ETDEWEB)

Gunsolus, Ian L.; Hu, Dehong; Mihai, Cosmin; Lohse, Samuel E.; Lee, Chang-Soo; Torelli, Marco; Hamers, Robert J.; Murphy, Catherine; Orr, Galya; Haynes, Christy L.

2014-01-01

A method to fluorescently stain the surfaces of both Gram-negative and Gram-positive bacterial cells compatible with super-resolution fluorescence microscopy is presented. This method utilizes a commercially-available fluorescent probe to label primary amines at the surface of the cell. We demonstrate efficient staining of two bacterial strains, the Gram-negative Shewanella oneidensis MR-1 and the Gram-positive Bacillus subtilis 168. Using structured illumination microscopy and stochastic optical reconstruction microscopy, which require high quantum yield or specialized dyes, we show that this staining method may be used to resolve the bacterial cell surface with sub-diffraction-limited resolution. We further use this method to identify localization patterns of nanomaterials, specifically cadmium selenide quantum dots, following interaction with bacterial cells.

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

International Nuclear Information System (INIS)

Mercouris, Theodoros; Nicolaides, Cleanthes A

2005-01-01

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

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

International Nuclear Information System (INIS)

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

2015-01-01

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

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

Energy Technology Data Exchange (ETDEWEB)

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

2015-04-13

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

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

International Nuclear Information System (INIS)

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

2013-01-01

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

Field enhancement of multiphoton induced luminescence processes in ZnO nanorods

Science.gov (United States)

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

2018-03-01

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

Two-photon emission and multiphoton absorption by atoms

International Nuclear Information System (INIS)

Mu, X.

1988-01-01

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

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

International Nuclear Information System (INIS)

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

2002-01-01

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

Technical parameters of vertical in vivo multiphoton microscopy: a critical evaluation of the flyscanning method

International Nuclear Information System (INIS)

Czekalla, C; Röwert-Huber, J; Darvin, M E; Lademann, J; Meinke, M C; Schönborn, K H; Markworth, S; Ulrich, M; Göppner, D; Gollnick, H

2015-01-01

The optical biopsy could be a quick and painless support or alternative to a punch biopsy. In this letter the first in vivo vertical wide field two photon microscopy (2PM) images of healthy volunteers are shown. The 2PM images are fused images of two photon excited auto fluorescence (AF) and second harmonic generation (SHG) signals given as false-color images of 200 μm  ×  7 mm in size. By using these two nonlinear effects, the epidermis can be easily distinguished from the dermis at a glance. The auto fluorescence provides cellular resolution of the epidermal cells, and elastin fibers are partly visible in the dermis. Collagen, visible by SHG signal, is the dominant structure in the dermis. As contact agent water was evaluated to increase the AF signal, especially in the deeper layers of epidermis and dermis. For further improvement any terminal hairs should be removed by shaving and by taking tape strips of the first five layers of the stratum corneum. The first images illustrated that young skin compared to aged skin shows remarkably different dermal elastin and collagen signals in the dermis. (letter)

Photoionization of excited molecular states using multiphoton excitation techniques

International Nuclear Information System (INIS)

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

1984-01-01

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

Multi-photon absorption limits to heralded single photon sources

Science.gov (United States)

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

2013-01-01

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

Multiphoton ionization photoelectron spectroscopy of xenon: Experiment and theory

International Nuclear Information System (INIS)

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

1988-11-01

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

Permutational symmetries for coincidence rates in multimode multiphotonic interferometry

Science.gov (United States)

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

2018-06-01

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

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

Science.gov (United States)

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

2014-01-01

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

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

Science.gov (United States)

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

2014-03-01

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

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

International Nuclear Information System (INIS)

Likhatov, P V; Telnov, D A

2009-01-01

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

Simultaneous multiphoton processes in the interaction of atoms with electromagnetic fields

International Nuclear Information System (INIS)

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

1984-01-01

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

Multiphoton production at high energies in the standard model. II

International Nuclear Information System (INIS)

Mahlon, G.

1993-01-01

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

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

Energy Technology Data Exchange (ETDEWEB)

Mette B. Gaarde

2009-08-28

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

Single and multi-photon events with missing energy in $e^+ e^-$ collisions at 161 GeV < $\\sqrt{s}$ < 172 GeV

CERN Document Server

Acciarri, M; Aguilar-Benítez, M; Ahlen, S P; Alcaraz, J; Alemanni, G; Allaby, James V; Aloisio, A; Alverson, G; Alviggi, M G; Ambrosi, G; Anderhub, H; Andreev, V P; Angelescu, T; Anselmo, F; Arefev, A; Azemoon, T; Aziz, T; Bagnaia, P; Baksay, L; Banerjee, S; Banerjee, Sw; Banicz, K; Barczyk, A; Barillère, R; Barone, L; Bartalini, P; Baschirotto, A; Basile, M; Battiston, R; Bay, A; Becattini, F; Becker, U; Behner, F; Berdugo, J; Berges, P; Bertucci, B; Betev, B L; Bhattacharya, S; Biasini, M; Biland, A; Bilei, G M; Blaising, J J; Blyth, S C; Bobbink, Gerjan J; Böck, R K; Böhm, A; Boldizsar, L; Borgia, B; Bourilkov, D; Bourquin, Maurice; Braccini, S; Branson, J G; Brigljevic, V; Brock, I C; Buffini, A; Buijs, A; Burger, J D; Burger, W J; Busenitz, J K; Button, A M; Cai, X D; Campanelli, M; Capell, M; Cara Romeo, G; Carlino, G; Cartacci, A M; Casaus, J; Castellini, G; Cavallari, F; Cavallo, N; Cecchi, C; Cerrada-Canales, M; Cesaroni, F; Chamizo-Llatas, M; Chang, Y H; Chaturvedi, U K; Chekanov, S V; Chemarin, M; Chen, A; Chen, G; Chen, G M; Chen, H F; Chen, H S; Chéreau, X J; Chiefari, G; Chien, C Y; Cifarelli, Luisa; Cindolo, F; Civinini, C; Clare, I; Clare, R; Cohn, H O; Coignet, G; Colijn, A P; Colino, N; Commichau, V; Costantini, S; Cotorobai, F; de la Cruz, B; Csilling, Akos; Dai, T S; D'Alessandro, R; De Asmundis, R; Degré, A; Deiters, K; Della Volpe, D; Denes, P; De Notaristefani, F; DiBitonto, Daryl; Diemoz, M; Van Dierendonck, D N; Di Lodovico, F; Dionisi, C; Dittmar, Michael; Dominguez, A; Doria, A; Dova, M T; Duchesneau, D; Duinker, P; Durán, I; Dutta, S; Easo, S; Efremenko, Yu V; El-Mamouni, H; Engler, A; Eppling, F J; Erné, F C; Ernenwein, J P; Extermann, Pierre; Fabre, M; Faccini, R; Falciano, S; Favara, A; Fay, J; Fedin, O; Felcini, Marta; Fenyi, B; Ferguson, T; Ferroni, F; Fesefeldt, H S; Fiandrini, E; Field, J H; Filthaut, Frank; Fisher, P H; Fisk, I; Forconi, G; Fredj, L; Freudenreich, Klaus; Furetta, C; Galaktionov, Yu; Ganguli, S N; García-Abia, P; Gau, S S; Gentile, S; Gheordanescu, N; Giagu, S; Goldfarb, S; Goldstein, J; Gong, Z F; Gougas, Andreas; Gratta, Giorgio; Grünewald, M W; Gupta, V K; Gurtu, A; Gutay, L J; Hartmann, B; Hasan, A; Hatzifotiadou, D; Hebbeker, T; Hervé, A; Van Hoek, W C; Hofer, H; Hong, S J; Hoorani, H; Hou, S R; Hu, G; Innocente, Vincenzo; Jenkes, K; Jin, B N; Jones, L W; de Jong, P; Josa-Mutuberria, I; Kasser, A; Khan, R A; Kamrad, D; Kamyshkov, Yu A; Kapustinsky, J S; Karyotakis, Yu; Kaur, M; Kienzle-Focacci, M N; Kim, D; Kim, D H; Kim, J K; Kim, S C; Kim, Y G; Kinnison, W W; Kirkby, A; Kirkby, D; Kirkby, Jasper; Kiss, D; Kittel, E W; Klimentov, A; König, A C; Kopp, A; Korolko, I; Koutsenko, V F; Krämer, R W; Krenz, W; Kunin, A; Ladrón de Guevara, P; Laktineh, I; Landi, G; Lapoint, C; Lassila-Perini, K M; Laurikainen, P; Lebeau, M; Lebedev, A; Lebrun, P; Lecomte, P; Lecoq, P; Le Coultre, P; Le Goff, J M; Leiste, R; Leonardi, E; Levchenko, P M; Li Chuan; Lin, C H; Lin, W T; Linde, Frank L; Lista, L; Liu, Z A; Lohmann, W; Longo, E; Lu, W; Lü, Y S; Lübelsmeyer, K; Luci, C; Luckey, D; Luminari, L; Lustermann, W; Ma Wen Gan; Maity, M; Majumder, G; Malgeri, L; Malinin, A; Maña, C; Mangeol, D J J; Mangla, S; Marchesini, P A; Marin, A; Martin, J P; Marzano, F; Massaro, G G G; McNally, D; McNeil, R R; Mele, S; Merola, L; Meschini, M; Metzger, W J; Von der Mey, M; Mi, Y; Mihul, A; Van Mil, A J W; Mirabelli, G; Mnich, J; Molnár, P; Monteleoni, B; Moore, R; Morganti, S; Moulik, T; Mount, R; Müller, S; Muheim, F; Muijs, A J M; Nahn, S; Napolitano, M; Nessi-Tedaldi, F; Newman, H; Niessen, T; Nippe, A; Nisati, A; Nowak, H; Oh, Yu D; Opitz, H; Organtini, G; Ostonen, R; Palomares, C; Pandoulas, D; Paoletti, S; Paolucci, P; Park, H K; Park, I H; Pascale, G; Passaleva, G; Patricelli, S; Paul, T; Pauluzzi, M; Paus, C; Pauss, Felicitas; Peach, D; Pei, Y J; Pensotti, S; Perret-Gallix, D; Petersen, B; Petrak, S; Pevsner, A; Piccolo, D; Pieri, M; Pinto, J C; Piroué, P A; Pistolesi, E; Plyaskin, V; Pohl, M; Pozhidaev, V; Postema, H; Produit, N; Prokofev, D; Prokofiev, D O; Rahal-Callot, G; Raja, N; Rancoita, P G; Rattaggi, M; Raven, G; Razis, P A; Read, K; Ren, D; Rescigno, M; Reucroft, S; Van Rhee, T; Riemann, S; Riles, K; Robohm, A; Rodin, J; Roe, B P; Romero, L; Rosier-Lees, S; Rosselet, P; Van Rossum, W; Roth, S; Rubio, Juan Antonio; Ruschmeier, D; Rykaczewski, H; Salicio, J; Sánchez, E; Sanders, M P; Sarakinos, M E; Sarkar, S; Sassowsky, M; Schäfer, C; Shchegelskii, V; Schmidt-Kärst, S; Schmitz, D; Schmitz, P; Scholz, N; Schopper, Herwig Franz; Schotanus, D J; Schwenke, J; Schwering, G; Sciacca, C; Sciarrino, D; Servoli, L; Shevchenko, S; Shivarov, N; Shoutko, V; Shukla, J; Shumilov, E; Shvorob, A V; Siedenburg, T; Son, D; Sopczak, André; Smith, B; Spillantini, P; Steuer, M; Stickland, D P; Stone, A; Stone, H; Stoyanov, B; Strässner, A; Strauch, K; Sudhakar, K; Sultanov, G G; Sun, L Z; Susinno, G F; Suter, H; Swain, J D; Tang, X W; Tauscher, Ludwig; Taylor, L; Ting, Samuel C C; Ting, S M; Tonutti, M; Tonwar, S C; Tóth, J; Tully, C; Tuchscherer, H; Tung, K L; Uchida, Y; Ulbricht, J; Uwer, U; Valente, E; Van de Walle, R T; Vesztergombi, G; Vetlitskii, I; Viertel, Gert M; Vivargent, M; Völkert, R; Vogel, H; Vogt, H; Vorobev, I; Vorobyov, A A; Vorvolakos, A; Wadhwa, M; Wallraff, W; Wang, J C; Wang, X L; Wang, Z M; Weber, A; Wittgenstein, F; Wu, S X; Wynhoff, S; Xu, J; Xu, Z Z; Yang, B Z; Yang, C G; Yao, X Y; Ye, J B; Yeh, S C; You, J M; Zalite, A; Zalite, Yu; Zemp, P; Zeng, Y; Zhang, Z; Zhang, Z P; Zhou, B; Zhu, G Y; Zhu, R Y; Zichichi, Antonino; Ziegler, F

1997-01-01

A search for single and multi-photon events with missing energy is performed using data collected at centre-of-mass energies between 161 GeV and 172 GeV for a total of 20.9 pb$^{-1}$ of integrated luminosity. The results obtained are used to derive the value for the $\

Reproducibility in light microscopy: Maintenance, standards and SOPs.

Science.gov (United States)

Deagle, Rebecca C; Wee, Tse-Luen Erika; Brown, Claire M

2017-08-01

Light microscopy has grown to be a valuable asset in both the physical and life sciences. It is a highly quantitative method available in individual research laboratories and often centralized in core facilities. However, although quantitative microscopy is becoming a customary tool in research, it is rarely standardized. To achieve accurate quantitative microscopy data and reproducible results, three levels of standardization must be considered: (1) aspects of the microscope, (2) the sample, and (3) the detector. The accuracy of the data is only as reliable as the imaging system itself, thereby imposing the need for routine standard performance testing. Depending on the task some maintenance procedures should be performed once a month, some before each imaging session, while others conducted annually. This text should be implemented as a resource for researchers to integrate with their own standard operating procedures to ensure the highest quality quantitative microscopy data. Copyright © 2017. Published by Elsevier Ltd.

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

Energy Technology Data Exchange (ETDEWEB)

Lee, Yuan T.; Shen, Y. Ron

1980-06-01

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

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

International Nuclear Information System (INIS)

Kimman, J.T.N.

1984-01-01

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

Photoionization of excited molecular states using multiphoton excitation techniques

International Nuclear Information System (INIS)

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

1984-01-01

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

National Center for Electron Microscopy users' guide

International Nuclear Information System (INIS)

1987-01-01

The National Center for Electron Microscopy (NCEM) in the Materials and Molecular Research Division of the Lawrence Berkeley Laboratory is a high voltage electron microscope facility for ultra-high resolution or dynamic in-situ studies. This guide describes the instruments and their specifications, support instrumentation, and user policies. Advice as to travel and accommodations is provided in the guide. (FI)

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

Directory of Open Access Journals (Sweden)

Adan Li

2014-01-01

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

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

International Nuclear Information System (INIS)

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

2012-01-01

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

Simulation of multi-photon emission isotopes using time-resolved SimSET multiple photon history generator

Energy Technology Data Exchange (ETDEWEB)

Chiang, Chih-Chieh; Lin, Hsin-Hon; Lin, Chang-Shiun; Chuang, Keh-Shih [Department of Biomedical Engineering and Environmental Sciences, National Tsing-HuaUniversity, Hsinchu, Taiwan (China); Jan, Meei-Ling [Health Physics Division, Institute of Nuclear Energy Research, Atomic Energy Council, Taoyuan, Taiwan (China)

2015-07-01

Abstract-Multiple-photon emitters, such as In-111 or Se-75, have enormous potential in the field of nuclear medicine imaging. For example, Se-75 can be used to investigate the bile acid malabsorption and measure the bile acid pool loss. The simulation system for emission tomography (SimSET) is a well-known Monte Carlo simulation (MCS) code in nuclear medicine for its high computational efficiency. However, current SimSET cannot simulate these isotopes due to the lack of modeling of complex decay scheme and the time-dependent decay process. To extend the versatility of SimSET for simulation of those multi-photon emission isotopes, a time-resolved multiple photon history generator based on SimSET codes is developed in present study. For developing the time-resolved SimSET (trSimSET) with radionuclide decay process, the new MCS model introduce new features, including decay time information and photon time-of-flight information, into this new code. The half-life of energy states were tabulated from the Evaluated Nuclear Structure Data File (ENSDF) database. The MCS results indicate that the overall percent difference is less than 8.5% for all simulation trials as compared to GATE. To sum up, we demonstrated that time-resolved SimSET multiple photon history generator can have comparable accuracy with GATE and keeping better computational efficiency. The new MCS code is very useful to study the multi-photon imaging of novel isotopes that needs the simulation of lifetime and the time-of-fight measurements. (authors)

Probing temporal aspects of high-order harmonic pulses via multi-colour, multi-photon ionization processes

Energy Technology Data Exchange (ETDEWEB)

Mauritsson, J [Department of Physics and Astronomy, Louisiana State University, Baton Rouge, LA 70803-4001 (United States); Johnsson, P [Department of Physics, Lund Institute of Technology, PO Box 118, SE-22100 Lund (Sweden); Lopez-Martens, R [Department of Physics, Lund Institute of Technology, PO Box 118, SE-22100 Lund (Sweden); Varju, K [Department of Physics, Lund Institute of Technology, PO Box 118, SE-22100 Lund (Sweden); L' Huillier, A [Department of Physics, Lund Institute of Technology, PO Box 118, SE-22100 Lund (Sweden); Gaarde, M B [Department of Physics and Astronomy, Louisiana State University, Baton Rouge, LA 70803-4001 (United States); Schafer, K J [Department of Physics and Astronomy, Louisiana State University, Baton Rouge, LA 70803-4001 (United States)

2005-07-14

High-order harmonics generated through the interaction of atoms and strong laser fields are a versatile, laboratory-scale source of extreme ultraviolet (XUV) radiation on a femtosecond or even attosecond time-scale. In order to be a useful experimental tool, however, this radiation has to be well characterized, both temporally and spectrally. In this paper we discuss how multi-photon, multi-colour ionization processes can be used to completely characterize either individual harmonics or attosecond pulse trains. In particular, we discuss the influence of the intensity and duration of the probe laser, and how these parameters effect the accuracy of the XUV characterization.

Probing temporal aspects of high-order harmonic pulses via multi-colour, multi-photon ionization processes

International Nuclear Information System (INIS)

Mauritsson, J; Johnsson, P; Lopez-Martens, R; Varju, K; L'Huillier, A; Gaarde, M B; Schafer, K J

2005-01-01

High-order harmonics generated through the interaction of atoms and strong laser fields are a versatile, laboratory-scale source of extreme ultraviolet (XUV) radiation on a femtosecond or even attosecond time-scale. In order to be a useful experimental tool, however, this radiation has to be well characterized, both temporally and spectrally. In this paper we discuss how multi-photon, multi-colour ionization processes can be used to completely characterize either individual harmonics or attosecond pulse trains. In particular, we discuss the influence of the intensity and duration of the probe laser, and how these parameters effect the accuracy of the XUV characterization

Resonance enhanced multiphoton ionization spectra of molecules and molecular fragments. Annual progress report, March 1992 - February 1993

International Nuclear Information System (INIS)

1993-01-01

In this report, the author will review the progress made in his studies of ion rotational distributions resulting from resonance enhanced multiphoton ionization of excited electronic states and from single-photon ionization of ground electronic states of jet-cooled molecules by coherent VUV and XUV radiation. To do so he will select a few examples from his studies which serve to highlight his progress and to identify the background and significance of the specific spectral features and systems he has chosen to study

Nano, Queensland and cryo-electron microscopy

International Nuclear Information System (INIS)

McDowall, A.W.

2002-01-01

Full text: In a recent review the authors, Wolfgang Baumeister and Alasdair Steven wrote, '....there is immense opportunity for Cryo-EM, especially as boosted by merging crystallographic structures of individual subunits into moderate resolution Cryo-EM density maps of whole complexes. Electron tomography has now advanced to the point where it is a realistic goal to glimpse molecular machines operating inside cells....' This statement gives testament to the advances made over the past 25 years by many labs around the world to the area of microscopy referred to as Cryo-EM and related 3-D computing technologies. Australian scientific societies have been eager followers of this progress and heard first hand of the new developments in the field at the 1984 ACEM-8 (2). Since those early days the ACEM and other Australian/NZ societies have sponsored numerous researchers and workshops in the field of Cryo-EM to their conferences, Helin Sabil, Wah Chiu, Ron Milligan, Richard Henderson and Werner Kuhlbrandt to name only a few. These visits have stimulated a desire from Australian/NZ researchers to establish collaborations and access to prominent labs in the USA and Europe, where the means and knowledge to provide Cryo EM and 3D reconstruction technology for studying macromolecular complexes is well established. However, Australia has not been backward in seeking to provide its home research community with access to a base in biological molecular microscopy and electron crystallography technology. Since the last ACEM we have seen the emergence of a number of crucial factors, which will make the establishment of a national research facility in this field an operational reality in early 2003. Well publicized is the development of Australia's newest and perhaps most unique research institute, the institute for Molecular Bioscience (IMB) to open at the University of Queensland (UQ) in 2002. The IMB will be the platform for a new research group in advanced computational 3D

An Automated System for the Control of, and Data Acquisition from Multiphoton Ionization and Fluorescence Lifetime Measurements.

Science.gov (United States)

1986-09-01

Quanta- Ray company , which also supplied the laser used for the multiphoton work. The, burner was mounted on a translator stage from Velmex, Inc...and no longer exists as a process in the system. When the user analysis program has completed, the lifetime program is again automatically re-started...KCHAR) RETURN 100 FORMAT(I3) 101 FORMAT(F7.2) END SUBROUTINE LAB4 FODA SE"oteD C This routine puts the label "INTEGRAL FROM DATA SET" on the MDP C screen

Detection of malaria parasites by microscopy and rapid diagnostic ...

African Journals Online (AJOL)

The effectiveness of Rapid Diagnostic Test Kit (RDT) was compared with microscopy for the evaluation of malaria infection in children and pregnant women attending two selected health facilities in Lagos State, south-western, Nigeria. A total of 482 patients comprising 252 pregnant women (mean age: 26.86±4.46 years) ...

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

International Nuclear Information System (INIS)

Chien, R.; Sogard, M.R.

1990-01-01

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

Multi-photon resonant effects in strong-field ionization: origin of the dip in experimental longitudinal momentum distributions

International Nuclear Information System (INIS)

Alnaser, A S; Maharjan, C M; Wang, P; Litvinyuk, I V

2006-01-01

We studied ionization of neon and argon by intense linearly polarized femtosecond laser pulses of different wavelengths (400 nm and 800 nm) and peak intensities, and by measuring momentum distributions of singly charged positive ions in the direction parallel to laser polarization. For Ne the momentum distributions exhibited a characteristic dip at zero momentum at 800 nm and a complex multipeak structure at 400 nm. Similarly, for Ar the momentum distributions evolved from a complex multipeak structure with a pronounced dip in the centre at 400 nm, to a smooth distribution characteristic of pure tunneling ionization (800 nm, high intensities). In the intermediate regime (800 nm, medium to low intensities), for both atoms we observed recoil ion momentum distributions modulated by quasi-periodic structures usually seen in the photoelectron energy spectra in a multi-photon regime (ATI spectra). Ne did show a characteristic 'dip' at low momentum, while the longitudinal momentum distribution for Ar exhibited a spike at zero momentum instead. The spectra did dramatically change at 400 nm, where both ions show the pronounced dip near zero momentum. Based on our results, we conclude that the structures observed in Ne and Ar momentum distributions reflect the specifics of atomic structure of the two targets and should not be attributed to effects of electron recollision, as was suggested earlier. Instead, as our results indicate, they are due to the effects of multi-photon resonant enhancement of strong-field ionization. (letter to the editor)

Multiphoton Ionization Detection in Collinear Laser Spectroscopy of Isolde Beams

CERN Multimedia

2002-01-01

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

A hard X-ray nanoprobe beamline for nanoscale microscopy

Energy Technology Data Exchange (ETDEWEB)

Winarski, Robert P., E-mail: winarski@anl.gov; Holt, Martin V. [Argonne National Laboratory, 9700 South Cass Avenue, Argonne, IL 60441 (United States); Rose, Volker [Argonne National Laboratory, 9700 South Cass Avenue, Argonne, IL 60441 (United States); Fuesz, Peter; Carbaugh, Dean; Benson, Christa; Shu, Deming; Kline, David; Stephenson, G. Brian; McNulty, Ian [Argonne National Laboratory, 9700 South Cass Avenue, Argonne, IL 60441 (United States); Maser, Jörg [Argonne National Laboratory, 9700 South Cass Avenue, Argonne, IL 60441 (United States)

2012-11-01

The Hard X-ray Nanoprobe Beamline is a precision platform for scanning probe and full-field microscopy with 3–30 keV X-rays. A combination of high-stability X-ray optics and precision motion sensing and control enables detailed studies of the internal features of samples with resolutions approaching 30 nm. The Hard X-ray Nanoprobe Beamline (or Nanoprobe Beamline) is an X-ray microscopy facility incorporating diffraction, fluorescence and full-field imaging capabilities designed and operated by the Center for Nanoscale Materials and the Advanced Photon Source at Sector 26 of the Advanced Photon Source at Argonne National Laboratory. This facility was constructed to probe the nanoscale structure of biological, environmental and material sciences samples. The beamline provides intense focused X-rays to the Hard X-ray Nanoprobe (or Nanoprobe) which incorporates Fresnel zone plate optics and a precision laser sensing and control system. The beamline operates over X-ray energies from 3 to 30 keV, enabling studies of most elements in the periodic table, with a particular emphasis on imaging transition metals.

Collaborative Initiative in Biomedical Imaging to Study Complex Diseases

Energy Technology Data Exchange (ETDEWEB)

Lin, Weili [The University of North Carolina at Chapel Hill; Fiddy, Michael A. [The University of North Carolina at Charlotte

2012-03-31

The work reported addressed these topics: Fluorescence imaging; Optical coherence tomography; X-ray interferometer/phase imaging system; Quantitative imaging from scattered fields, Terahertz imaging and spectroscopy; and Multiphoton and Raman microscopy.

Microsphere imaging with confocal microscopy and two photon microscopy

International Nuclear Information System (INIS)

Chun, Hyung Su; An, Kyung Won; Lee, Jai Hyung

2002-01-01

We have acquired images of polystyrene and fused-silica microsphere by using conventional optical microscopy, confocal microscopy and two-photon microscopy, and performed comparative analysis of these images. Different from conventional optical microscopy, confocal and two-photon microscopy had good optical sectioning capability. In addition, confocal microscopy and two-photon microscopy had better lateral resolution than conventional optical microscopy. These results are attributed to confocality and nonlinearity of confocal microscopy and two photon microscopy, respectively.

Multiphoton ionization for hydrogen plasma diagnostics

International Nuclear Information System (INIS)

Bonnie, J.H.M.

1987-01-01

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

Paracheck-PF ® test Versus Microscopy in the Diagnosis of ...

African Journals Online (AJOL)

Background: Malaria is a major cause of morbidity and mortality in Ethiopia. Rapid diagnostic tests such as Paracheck Pf are the major tools for falciparum malaria diagnosis as an alternative to microscopy in peripheral health facilities. The objective of this study was to evaluate the sensitivity and specificity of Paracheck Pf ...

Theory of the effect of third-harmonic generation on three-photon resonantly enhanced multiphoton ionization in focused beams

International Nuclear Information System (INIS)

Payne, M.G.; Garrett, W.R.

1983-01-01

Multiphoton ionization in the region near a three-photon resonance is treated for focused, plane-polarized Gaussian beams with diffraction-limited beam divergence. In this situation, a third-harmonic field is generated within the laser beam. At, and very near, three-photon resonance the driving rate for the upper-state probability amplitude due to one-photon absorption of third-harmonic light becomes nearly equal to the corresponding three-photon rate due to the laser field, but these effects are 180 0 out of phase. As a consequence of this cancellation between two pumping terms, the three-photon resonance line essentially disappears at moderate concentrations and the observed ionization has a line shape that is close to the phase-matching curve for third-harmonic generation. The ionization signal, near but not on the resonance, is due almost entirely to absorption of third-harmonic photons plus other laser photons; three-photon resonantly enhanced multiphoton ionization by the laser is much weaker. This is particularly true on the blue side of the three-photon resonance at detunings where phase matching occurs. The problem is treated quite generally with predictions of the full line shape for n-photon ionization and third-harmonic light generation near three-photon resonance, including the rather strong influences of positively dispersive buffer gases. We also show that the cancellation between the one-photon and the three-photon process is partially spoiled in the presence of a counterpropagating beam at the same frequency

The future of the SIRAD SEE facility Ion-Electron Emission Microscopy

CERN Document Server

Wyss, J; Kaminski, A; Magalini, A; Nigro, M; Pantano, D; Sedhykh, S

2002-01-01

The SIRAD facility is dedicated to radiation damage studies on semiconductor detectors, electronic devices and systems, using proton and ion beams delivered by a 15 MV tandem accelerator. It is routinely used by groups involved in detector development for elementary particle physics, electronic device physics and space applications. In particular, Single Event Effect studies are very important to the latter two activities. Presently, the facility can only characterize the global sensitivity of a device or system to single ion impacts. To map out the sensitivity of a device with micrometric resolution, following an idea developed at SANDIA, we will implement an Ion-Electron Emission Microscope (IEEM) to reconstruct the X,Y and time coordinates of an impacting energetic ion by imaging the secondary electrons emitted by the sample using a standard emission electron microscope and position sensitive detector system. After describing typical Single Event Effect activities at SIRAD we will discuss the basic princip...

Multiphoton ionization of (Xe)/sub n/ and (NO)/sub n/ clusters using a picosecond laser

International Nuclear Information System (INIS)

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

1989-01-01

In an effort to extend the application of multiphoton ionization (MPI) spectroscopy to the study of weakly bound systems, we have begun a systematic investigation of picosecond MPI in van der Waals molecules and clusters. To our knowledge no previous picosecond MPI studies of weakly bound systems have been reported. We present here results of picosecond MPI of Xe/sub n/(n = 1-20) and (NO)/sub n/(n = 1-4) clusters. Previous MPI studies using nanosecond lasers have not detected the NO cluster series, presumably because of fast dissociation channels. The use of high peak-power allows resonant and non-resonant photon absorption to the ionization limit to compete effectively with fast dissociative processes. 10 refs., 2 figs

Dissecting multi-photon resonances at the large hadron collider

Energy Technology Data Exchange (ETDEWEB)

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

2017-09-15

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

Energetics of the rearrangement of neutral and ionized perfluorocyclopropane to perfluoropropylene. Use of infrared multiphoton dissociation spectra to identify structural isomers of molecular ions

International Nuclear Information System (INIS)

Bomse, D.S.; Berman, D.W.; Beauchamp, J.L.

1981-01-01

Infrared photodissociation spectroscopy is used to compare the structure of gas-phase C 3 F 6 + ions obtained by electron-impact ionization of two isomeric precursors: perfluoropropylene and perfluorocyclopropane. Photodissociation spectra are obtained by observing the extent of multiphoton dissociation as the CO 2 laser is tuned across the 925 to 1080 cm -1 wavelength range. Ions are formed, stored, and detected with the use of techniques of ion cyclotron resonance spectroscopy. Infrared multiphoton excitation is effected by using low-power, continuous-wave laser radiation. The fingerprint spectrum of the molecular ion of perfluorocyclopropane is identical with that obtained from perfluoropropylene, indicating rearrangement of the former to the latter. Photodissociation kinetics indicate that the entire perfluorocyclopropane molecular ion population isomerizes to the more stable perfluoropropylene structure. Thermochemistry of C 3 F 6 and C 3 F 6 + isomers is discussed. Comparisons are made with the analogous C 3 H 6 system. Photoionization mass spectroscopy results yield ΔH/sub f/(c-C 3 F 6 ) = -233.8 kcal/mol. 4 figures

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

International Nuclear Information System (INIS)

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

1991-07-01

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

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

Science.gov (United States)

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

2018-03-01

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

Impulsive IR-multiphoton dissociation of acrolein: observation of non-statistical product vibrational excitation in CO ( v=1-12) by time resolved IR fluorescence spectroscopy

Science.gov (United States)

Chowdhury, P. K.

2000-10-01

On IR-multiphoton excitation, vibrationally highly excited acrolein molecules undergo concerted dissociation generating CO and ethylene. The vibrationally excited products, CO and ethylene, are detected immediately following the CO 2 laser pulse by observing IR fluorescence at 4.7 and 3.2 μm, respectively. The nascent CO is formed with significant vibrational excitation, with a Boltzmann population distribution for v=1-12 levels corresponding to T v=12 950±50 K. The average vibrational energy in the product CO is found to be 26 kcal mol -1, in contrast to its statistical share of 5 kcal mol -1, available from the product energy distribution. The nascent vibrationally excited ethylene either dissociates by absorbing further infrared laser photons from the tail of the CO 2 laser pulse or relaxes by collisional deactivation. Ethylene IR-fluorescence excitation spectrum showed a structure in the quasi-continuum, with a facile resonance at 10.53 μm corresponding to the 10P(14) CO 2 laser line, which explains the higher acetylene yield observed at a higher pressure. A hydrogen atom transfer mechanism followed by C-C impulsive break in the acrolein transition state may be responsible for such non-statistical product energy distribution.

Subdiffraction Multicolor Imaging of the Nuclear Periphery with 3D Structured Illumination Microscopy

Science.gov (United States)

Schermelleh, Lothar; Carlton, Peter M.; Haase, Sebastian; Shao, Lin; Winoto, Lukman; Kner, Peter; Burke, Brian; Cardoso, M. Cristina; Agard, David A.; Gustafsson, Mats G. L.; Leonhardt, Heinrich; Sedat, John W.

2010-01-01

Fluorescence light microscopy allows multicolor visualization of cellular components with high specificity, but its utility has until recently been constrained by the intrinsic limit of spatial resolution. We applied three-dimensional structured illumination microscopy (3D-SIM) to circumvent this limit and to study the mammalian nucleus. By simultaneously imaging chromatin, nuclear lamina, and the nuclear pore complex (NPC), we observed several features that escape detection by conventional microscopy. We could resolve single NPCs that colocalized with channels in the lamin network and peripheral heterochromatin. We could differentially localize distinct NPC components and detect double-layered invaginations of the nuclear envelope in prophase as previously seen only by electron microscopy. Multicolor 3D-SIM opens new and facile possibilities to analyze subcellular structures beyond the diffraction limit of the emitted light. PMID:18535242

All-optical bidirectional neural interfacing using hybrid multiphoton holographic optogenetic stimulation.

Science.gov (United States)

Paluch-Siegler, Shir; Mayblum, Tom; Dana, Hod; Brosh, Inbar; Gefen, Inna; Shoham, Shy

2015-07-01

Our understanding of neural information processing could potentially be advanced by combining flexible three-dimensional (3-D) neuroimaging and stimulation. Recent developments in optogenetics suggest that neurophotonic approaches are in principle highly suited for noncontact stimulation of network activity patterns. In particular, two-photon holographic optical neural stimulation (2P-HONS) has emerged as a leading approach for multisite 3-D excitation, and combining it with temporal focusing (TF) further enables axially confined yet spatially extended light patterns. Here, we study key steps toward bidirectional cell-targeted 3-D interfacing by introducing and testing a hybrid new 2P-TF-HONS stimulation path for accurate parallel optogenetic excitation into a recently developed hybrid multiphoton 3-D imaging system. The system is shown to allow targeted all-optical probing of in vitro cortical networks expressing channelrhodopsin-2 using a regeneratively amplified femtosecond laser source tuned to 905 nm. These developments further advance a prospective new tool for studying and achieving distributed control over 3-D neuronal circuits both in vitro and in vivo.

Design and implementation of a sensitive high-resolution nonlinear spectral imaging microscope

NARCIS (Netherlands)

Palero, Jonathan A.; Latouche, Gwendal; de Bruijn, Henriëtte S.; van der Ploeg van den Heuvel, Angélique; Sterenborg, Henricus J. C. M.; Gerritsen, Hans C.

2008-01-01

Live tissue nonlinear microscopy based on multiphoton autofluorescence and second harmonic emission originating from endogenous fluorophores and noncentrosymmetric-structured proteins is rapidly gaining interest in biomedical applications. The advantage of this technique includes high imaging

Real-time dynamic range and signal to noise enhancement in beam-scanning microscopy by integration of sensor characteristics, data acquisition hardware, and statistical methods

Science.gov (United States)

Kissick, David J.; Muir, Ryan D.; Sullivan, Shane Z.; Oglesbee, Robert A.; Simpson, Garth J.

2013-02-01

Despite the ubiquitous use of multi-photon and confocal microscopy measurements in biology, the core techniques typically suffer from fundamental compromises between signal to noise (S/N) and linear dynamic range (LDR). In this study, direct synchronous digitization of voltage transients coupled with statistical analysis is shown to allow S/N approaching the theoretical maximum throughout an LDR spanning more than 8 decades, limited only by the dark counts of the detector on the low end and by the intrinsic nonlinearities of the photomultiplier tube (PMT) detector on the high end. Synchronous digitization of each voltage transient represents a fundamental departure from established methods in confocal/multi-photon imaging, which are currently based on either photon counting or signal averaging. High information-density data acquisition (up to 3.2 GB/s of raw data) enables the smooth transition between the two modalities on a pixel-by-pixel basis and the ultimate writing of much smaller files (few kB/s). Modeling of the PMT response allows extraction of key sensor parameters from the histogram of voltage peak-heights. Applications in second harmonic generation (SHG) microscopy are described demonstrating S/N approaching the shot-noise limit of the detector over large dynamic ranges.

Conjugate adaptive optics with remote focusing in multiphoton microscopy

Science.gov (United States)

Tao, Xiaodong; Lam, Tuwin; Zhu, Bingzhao; Li, Qinggele; Reinig, Marc R.; Kubby, Joel

2018-02-01

The small correction volume for conventional wavefront shaping methods limits their application in biological imaging through scattering media. In this paper, we take advantage of conjugate adaptive optics (CAO) and remote focusing (CAORF) to achieve three-dimensional (3D) scanning through a scattering layer with a single correction. Our results show that the proposed system can provide 10 times wider axial field of view compared with a conventional conjugate AO system when 16,384 segments are used on a spatial light modulator. We demonstrate two-photon imaging with CAORF through mouse skull. The fluorescent microspheres embedded under the scattering layers can be clearly observed after applying the correction.

A new method using multiphoton imaging and morphometric analysis for differentiating chromophobe renal cell carcinoma and oncocytoma kidney tumors

Science.gov (United States)

Wu, Binlin; Mukherjee, Sushmita; Jain, Manu

2016-03-01

Distinguishing chromophobe renal cell carcinoma (chRCC) from oncocytoma on hematoxylin and eosin images may be difficult and require time-consuming ancillary procedures. Multiphoton microscopy (MPM), an optical imaging modality, was used to rapidly generate sub-cellular histological resolution images from formalin-fixed unstained tissue sections from chRCC and oncocytoma.Tissues were excited using 780nm wavelength and emission signals (including second harmonic generation and autofluorescence) were collected in different channels between 390 nm and 650 nm. Granular structure in the cell cytoplasm was observed in both chRCC and oncocytoma. Quantitative morphometric analysis was conducted to distinguish chRCC and oncocytoma. To perform the analysis, cytoplasm and granules in tumor cells were segmented from the images. Their area and fluorescence intensity were found in different channels. Multiple features were measured to quantify the morphological and fluorescence properties. Linear support vector machine (SVM) was used for classification. Re-substitution validation, cross validation and receiver operating characteristic (ROC) curve were implemented to evaluate the efficacy of the SVM classifier. A wrapper feature algorithm was used to select the optimal features which provided the best predictive performance in separating the two tissue types (classes). Statistical measures such as sensitivity, specificity, accuracy and area under curve (AUC) of ROC were calculated to evaluate the efficacy of the classification. Over 80% accuracy was achieved as the predictive performance. This method, if validated on a larger and more diverse sample set, may serve as an automated rapid diagnostic tool to differentiate between chRCC and oncocytoma. An advantage of such automated methods are that they are free from investigator bias and variability.

Correlative Stochastic Optical Reconstruction Microscopy and Electron Microscopy

Science.gov (United States)

Kim, Doory; Deerinck, Thomas J.; Sigal, Yaron M.; Babcock, Hazen P.; Ellisman, Mark H.; Zhuang, Xiaowei

2015-01-01

Correlative fluorescence light microscopy and electron microscopy allows the imaging of spatial distributions of specific biomolecules in the context of cellular ultrastructure. Recent development of super-resolution fluorescence microscopy allows the location of molecules to be determined with nanometer-scale spatial resolution. However, correlative super-resolution fluorescence microscopy and electron microscopy (EM) still remains challenging because the optimal specimen preparation and imaging conditions for super-resolution fluorescence microscopy and EM are often not compatible. Here, we have developed several experiment protocols for correlative stochastic optical reconstruction microscopy (STORM) and EM methods, both for un-embedded samples by applying EM-specific sample preparations after STORM imaging and for embedded and sectioned samples by optimizing the fluorescence under EM fixation, staining and embedding conditions. We demonstrated these methods using a variety of cellular targets. PMID:25874453

Multiphoton control of the 1,3-cyclohexadiene ring-opening reaction in the presence of competing solvent reactions.

Science.gov (United States)

Carroll, Elizabeth C; White, James L; Florean, Andrei C; Bucksbaum, Philip H; Sension, Roseanne J

2008-07-31

Although physical chemistry has often concentrated on the observation and understanding of chemical systems, the defining characteristic of chemistry remains the direction and control of chemical reactivity. Optical control of molecular dynamics, and thus of chemical reactivity provides a path to use photon energy as a smart reagent in a chemical system. In this paper, we discuss recent research in this field in the context of our studies of the multiphoton optical control of the photo-initiated ring-opening reaction of 1,3-cyclohexadiene (CHD) to form 1,3,5- cis-hexatriene (Z-HT). Closed-loop feedback and learning algorithms are able to identify pulses that increase the desired target state by as much as a factor of two. Mechanisms for control are discussed through the influence of the intensity dependence, the nonlinear power spectrum, and the projection of the pulses onto low orders of polynomial phase. Control measurements in neat solvents demonstrate that competing solvent fragmentation reactions must also be considered. In particular, multiphoton excitation of cyclohexane alone is capable of producing hexatriene. Statistical analyses of data sets obtained in learning algorithm searches in neat cyclohexane and for CHD in hexane and cyclohexane highlight the importance of linear and quadratic chirp, while demonstrating that the control features are not so easily defined. Higher order phase components are also important. On the basis of these results the involvement of low-frequency ground-state vibrational modes is proposed. When the population is transferred to the excited state, momentum along the torsional coordinate may keep the wave packet localized as it moves toward the conical intersections controlling the yield of Z-HT.

Signatures of tunneling and multiphoton ionization in the electron-momentum distributions of atoms by intense few-cycle laser pulses

International Nuclear Information System (INIS)

Wickenhauser, M.; Tong, X. M.; Arbo, D. G.; Burgdoerfer, J.; Lin, C. D.

2006-01-01

Electron-momentum distributions for above-threshold ionization of argon in a few-cycle, linearly polarized laser pulse are investigated. Spectral features characteristic of multiphoton as well as tunneling ionization coexist over a range of the Keldysh parameter γ in the transition regime γ∼1. Surprisingly, the simple strong-field approximation (SFA) is capable of reproducing the key features of the two-dimensional momentum distributions found in the full solution of the time-dependent Schroedinger equation, despite the fact that SFA is known to severely underestimate the total ionization probability

Infrared multiphoton dissociation of acrolein. Time-resolved observation of CO ( v = 1) IR emission at 4.7 μm

Science.gov (United States)

Chowdhury, P. K.; Rama Rao, K. V. S.; Mittal, J. P.

1994-02-01

In contrast to the photochemistry of electronically excited acrolein producing vinyl and formyl radicals via CC bond rupture, multiphoton vibrationally excited molecules undergo concerted dissociation generating CO and ethylene. Vibrational excitation in the CO product is detected immediately following the CO 2 laser pulse by observing IR emission at 4.7 μm. The decay of the IR emission was studied as a function of acrolein pressure. A vibrational-vibrational relaxation rate constant of CO ( v=1) by acrolein is found to be 1240 ± 200 Torr -1 s -1.

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

Energy Technology Data Exchange (ETDEWEB)

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

2014-07-01

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

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

Science.gov (United States)

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

2005-12-01

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

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

International Nuclear Information System (INIS)

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

2006-01-01

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

In-vivo nonlinear optical microscopy (NLOM) of epithelial-connective tissue interface (ECTI) reveals quantitative measures of neoplasia in hamster oral mucosa.

Science.gov (United States)

Pal, Rahul; Yang, Jinping; Ortiz, Daniel; Qiu, Suimin; Resto, Vicente; McCammon, Susan; Vargas, Gracie

2015-01-01

The epithelial-connective tissue interface (ECTI) plays an integral role in epithelial neoplasia, including oral squamous cell carcinoma (OSCC). This interface undergoes significant alterations due to hyperproliferating epithelium that supports the transformation of normal epithelium to precancers and cancer. We present a method based on nonlinear optical microscopy to directly assess the ECTI and quantify dysplastic alterations using a hamster model for oral carcinogenesis. Neoplastic and non-neoplastic normal mucosa were imaged in-vivo by both multiphoton autofluorescence microscopy (MPAM) and second harmonic generation microscopy (SHGM) to obtain cross-sectional reconstructions of the oral epithelium and lamina propria. Imaged sites were biopsied and processed for histopathological grading and measurement of ECTI parameters. An ECTI shape parameter was calculated based on deviation from the linear geometry (ΔLinearity) seen in normal mucosa was measured using MPAM-SHGM and histology. The ECTI was readily visible in MPAM-SHGM and quantitative shape analysis showed ECTI deformation in dysplasia but not in normal mucosa. ΔLinearity was significantly (p tissue with 87.9% sensitivity and 97.6% specificity, while calculations from histology provided 96.4% sensitivity and 85.7% specificity. Among other quantifiable architectural changes, a progressive statistically significant increase in epithelial thickness was seen with increasing grade of dysplasia. MPAM-SHGM provides new noninvasive ways for direct characterization of ECTI which may be used in preclinical studies to investigate the role of this interface in early transformation. Further development of the method may also lead to new diagnostic approaches to differentiate non-neoplastic tissue from precancers and neoplasia, possibly with other cellular and layer based indicators of abnormality.

Fluorescence microscopy.

Science.gov (United States)

Sanderson, Michael J; Smith, Ian; Parker, Ian; Bootman, Martin D

2014-10-01

Fluorescence microscopy is a major tool with which to monitor cell physiology. Although the concepts of fluorescence and its optical separation using filters remain similar, microscope design varies with the aim of increasing image contrast and spatial resolution. The basics of wide-field microscopy are outlined to emphasize the selection, advantages, and correct use of laser scanning confocal microscopy, two-photon microscopy, scanning disk confocal microscopy, total internal reflection, and super-resolution microscopy. In addition, the principles of how these microscopes form images are reviewed to appreciate their capabilities, limitations, and constraints for operation. © 2014 Cold Spring Harbor Laboratory Press.

Joint-dependent response to impact and implications for post-traumatic osteoarthritis

DEFF Research Database (Denmark)

Novakofski, K D; Berg, Lise Charlotte; Bronzini, I

2015-01-01

RELEVANCE: Understanding differences in the response to injury and potential susceptibility to OA can lead to the development of preventative or treatment strategies. KEY TERMS: Gene expression, cartilage injury, chondrocyte, multiphoton microscopy, cartilage biomechanical properties, PTOA. WHAT IS KNOWN...

Facility for low-temperature spin-polarized-scanning tunneling microscopy studies of magnetic/spintronic materials prepared in situ by nitride molecular beam epitaxy.

Science.gov (United States)

Lin, Wenzhi; Foley, Andrew; Alam, Khan; Wang, Kangkang; Liu, Yinghao; Chen, Tianjiao; Pak, Jeongihm; Smith, Arthur R

2014-04-01

Based on the interest in, as well as exciting outlook for, nitride semiconductor based structures with regard to electronic, optoelectronic, and spintronic applications, it is compelling to investigate these systems using the powerful technique of spin-polarized scanning tunneling microscopy (STM), a technique capable of achieving magnetic resolution down to the atomic scale. However, the delicate surfaces of these materials are easily corrupted by in-air transfers, making it unfeasible to study them in stand-alone ultra-high vacuum STM facilities. Therefore, we have carried out the development of a hybrid system including a nitrogen plasma assisted molecular beam epitaxy/pulsed laser epitaxy facility for sample growth combined with a low-temperature, spin-polarized scanning tunneling microscope system. The custom-designed molecular beam epitaxy growth system supports up to eight sources, including up to seven effusion cells plus a radio frequency nitrogen plasma source, for epitaxially growing a variety of materials, such as nitride semiconductors, magnetic materials, and their hetero-structures, and also incorporating in situ reflection high energy electron diffraction. The growth system also enables integration of pulsed laser epitaxy. The STM unit has a modular design, consisting of an upper body and a lower body. The upper body contains the coarse approach mechanism and the scanner unit, while the lower body accepts molecular beam epitaxy grown samples using compression springs and sample skis. The design of the system employs two stages of vibration isolation as well as a layer of acoustic noise isolation in order to reduce noise during STM measurements. This isolation allows the system to effectively acquire STM data in a typical lab space, which during its construction had no special and highly costly elements included, (such as isolated slabs) which would lower the environmental noise. The design further enables tip exchange and tip coating without

Facility for low-temperature spin-polarized-scanning tunneling microscopy studies of magnetic/spintronic materials prepared in situ by nitride molecular beam epitaxy

Energy Technology Data Exchange (ETDEWEB)

Lin, Wenzhi; Foley, Andrew; Alam, Khan; Wang, Kangkang; Liu, Yinghao; Chen, Tianjiao; Pak, Jeongihm; Smith, Arthur R., E-mail: smitha2@ohio.edu [Department of Physics and Astronomy, Nanoscale and Quantum Phenomena Institute, Ohio University, Athens, Ohio 45701 (United States)

2014-04-15

Based on the interest in, as well as exciting outlook for, nitride semiconductor based structures with regard to electronic, optoelectronic, and spintronic applications, it is compelling to investigate these systems using the powerful technique of spin-polarized scanning tunneling microscopy (STM), a technique capable of achieving magnetic resolution down to the atomic scale. However, the delicate surfaces of these materials are easily corrupted by in-air transfers, making it unfeasible to study them in stand-alone ultra-high vacuum STM facilities. Therefore, we have carried out the development of a hybrid system including a nitrogen plasma assisted molecular beam epitaxy/pulsed laser epitaxy facility for sample growth combined with a low-temperature, spin-polarized scanning tunneling microscope system. The custom-designed molecular beam epitaxy growth system supports up to eight sources, including up to seven effusion cells plus a radio frequency nitrogen plasma source, for epitaxially growing a variety of materials, such as nitride semiconductors, magnetic materials, and their hetero-structures, and also incorporating in situ reflection high energy electron diffraction. The growth system also enables integration of pulsed laser epitaxy. The STM unit has a modular design, consisting of an upper body and a lower body. The upper body contains the coarse approach mechanism and the scanner unit, while the lower body accepts molecular beam epitaxy grown samples using compression springs and sample skis. The design of the system employs two stages of vibration isolation as well as a layer of acoustic noise isolation in order to reduce noise during STM measurements. This isolation allows the system to effectively acquire STM data in a typical lab space, which during its construction had no special and highly costly elements included, (such as isolated slabs) which would lower the environmental noise. The design further enables tip exchange and tip coating without

Facility for low-temperature spin-polarized-scanning tunneling microscopy studies of magnetic/spintronic materials prepared in situ by nitride molecular beam epitaxy

International Nuclear Information System (INIS)

Lin, Wenzhi; Foley, Andrew; Alam, Khan; Wang, Kangkang; Liu, Yinghao; Chen, Tianjiao; Pak, Jeongihm; Smith, Arthur R.

2014-01-01

Based on the interest in, as well as exciting outlook for, nitride semiconductor based structures with regard to electronic, optoelectronic, and spintronic applications, it is compelling to investigate these systems using the powerful technique of spin-polarized scanning tunneling microscopy (STM), a technique capable of achieving magnetic resolution down to the atomic scale. However, the delicate surfaces of these materials are easily corrupted by in-air transfers, making it unfeasible to study them in stand-alone ultra-high vacuum STM facilities. Therefore, we have carried out the development of a hybrid system including a nitrogen plasma assisted molecular beam epitaxy/pulsed laser epitaxy facility for sample growth combined with a low-temperature, spin-polarized scanning tunneling microscope system. The custom-designed molecular beam epitaxy growth system supports up to eight sources, including up to seven effusion cells plus a radio frequency nitrogen plasma source, for epitaxially growing a variety of materials, such as nitride semiconductors, magnetic materials, and their hetero-structures, and also incorporating in situ reflection high energy electron diffraction. The growth system also enables integration of pulsed laser epitaxy. The STM unit has a modular design, consisting of an upper body and a lower body. The upper body contains the coarse approach mechanism and the scanner unit, while the lower body accepts molecular beam epitaxy grown samples using compression springs and sample skis. The design of the system employs two stages of vibration isolation as well as a layer of acoustic noise isolation in order to reduce noise during STM measurements. This isolation allows the system to effectively acquire STM data in a typical lab space, which during its construction had no special and highly costly elements included, (such as isolated slabs) which would lower the environmental noise. The design further enables tip exchange and tip coating without

Clinical coherent anti-Stokes Raman scattering and multiphoton tomography of human skin with a femtosecond laser and photonic crystal fiber

International Nuclear Information System (INIS)

Breunig, Hans Georg; Weinigel, Martin; Bückle, Rainer; Kellner-Höfer, Marcel; König, Karsten; Lademann, Jürgen; Darvin, Maxim E; Sterry, Wolfram

2013-01-01

We report on in vivo coherent anti-Stokes Raman scattering spectroscopy (CARS), two-photon fluorescence and second-harmonic-generation imaging on human skin with a novel multimodal clinical CARS/multiphoton tomograph. CARS imaging is realized by a combination of femtosecond pulses with broadband continuum pulses generated by a photonic crystal fiber. The images reveal the microscopic distribution of (i) non-fluorescent lipids, (ii) endogenous fluorophores and (iii) the collagen network inside the human skin in vivo with subcellular resolution. Examples of healthy as well as cancer-affected skin are presented. (letter)

Clinical coherent anti-Stokes Raman scattering and multiphoton tomography of human skin with a femtosecond laser and photonic crystal fiber

Science.gov (United States)

Breunig, Hans Georg; Weinigel, Martin; Bückle, Rainer; Kellner-Höfer, Marcel; Lademann, Jürgen; Darvin, Maxim E.; Sterry, Wolfram; König, Karsten

2013-02-01

We report on in vivo coherent anti-Stokes Raman scattering spectroscopy (CARS), two-photon fluorescence and second-harmonic-generation imaging on human skin with a novel multimodal clinical CARS/multiphoton tomograph. CARS imaging is realized by a combination of femtosecond pulses with broadband continuum pulses generated by a photonic crystal fiber. The images reveal the microscopic distribution of (i) non-fluorescent lipids, (ii) endogenous fluorophores and (iii) the collagen network inside the human skin in vivo with subcellular resolution. Examples of healthy as well as cancer-affected skin are presented.

Open-ended response theory with polarizable embedding: multiphoton absorption in biomolecular systems.

Science.gov (United States)

Steindal, Arnfinn Hykkerud; Beerepoot, Maarten T P; Ringholm, Magnus; List, Nanna Holmgaard; Ruud, Kenneth; Kongsted, Jacob; Olsen, Jógvan Magnus Haugaard

2016-10-12

We present the theory and implementation of an open-ended framework for electric response properties at the level of Hartree-Fock and Kohn-Sham density functional theory that includes effects from the molecular environment modeled by the polarizable embedding (PE) model. With this new state-of-the-art multiscale functionality, electric response properties to any order can be calculated for molecules embedded in polarizable atomistic molecular environments ranging from solvents to complex heterogeneous macromolecules such as proteins. In addition, environmental effects on multiphoton absorption (MPA) properties can be studied by evaluating single residues of the response functions. The PE approach includes mutual polarization effects between the quantum and classical parts of the system through induced dipoles that are determined self-consistently with respect to the electronic density. The applicability of our approach is demonstrated by calculating MPA strengths up to four-photon absorption for the green fluorescent protein. We show how the size of the quantum region, as well as the treatment of the border between the quantum and classical regions, is crucial in order to obtain reliable MPA predictions.

Time-resolved photoion imaging spectroscopy: Determining energy distribution in multiphoton absorption experiments

Science.gov (United States)

Qian, D. B.; Shi, F. D.; Chen, L.; Martin, S.; Bernard, J.; Yang, J.; Zhang, S. F.; Chen, Z. Q.; Zhu, X. L.; Ma, X.

2018-04-01

We propose an approach to determine the excitation energy distribution due to multiphoton absorption in the case of excited systems following decays to produce different ion species. This approach is based on the measurement of the time-resolved photoion position spectrum by using velocity map imaging spectrometry and an unfocused laser beam with a low fluence and homogeneous profile. Such a measurement allows us to identify the species and the origin of each ion detected and to depict the energy distribution using a pure Poisson's equation involving only one variable which is proportional to the absolute photon absorption cross section. A cascade decay model is used to build direct connections between the energy distribution and the probability to detect each ionic species. Comparison between experiments and simulations permits the energy distribution and accordingly the absolute photon absorption cross section to be determined. This approach is illustrated using C60 as an example. It may therefore be extended to a wide variety of molecules and clusters having decay mechanisms similar to those of fullerene molecules.

Optical characterization of directly deposited graphene on a dielectric substrate

DEFF Research Database (Denmark)

Kaplas, Tommi; Karvonen, Lasse; Ahmadi, Sepehr

2016-01-01

By using scanning multiphoton microscopy we compare the nonlinear optical properties of the directly deposited and transferred to the dielectric substrate graphene. The direct deposition of graphene on oxidized silicon wafer was done by utilizing sacrificial copper catalyst film. We demonstrate...

In Vivo Microscopy Reveals Extensive Embedding of Capillaries within the Sarcolemma of Skeletal Muscle Fibers

Science.gov (United States)

Glancy, Brian; Hsu, Li-Yueh; Dao, Lam; Bakalar, Matthew; French, Stephanie; Chess, David J.; Taylor, Joni L.; Picard, Martin; Aponte, Angel; Daniels, Mathew P.; Esfahani, Shervin; Cushman, Samuel; Balaban, Robert S.

2013-01-01

Objective To provide insight into mitochondrial function in vivo, we evaluated the 3D spatial relationship between capillaries, mitochondria, and muscle fibers in live mice. Methods 3D volumes of in vivo murine Tibialis anterior muscles were imaged by multi-photon microscopy (MPM). Muscle fiber type, mitochondrial distribution, number of capillaries, and capillary-to-fiber contact were assessed. The role of myoglobin-facilitated diffusion was examined in myoglobin knockout mice. Distribution of GLUT4 was also evaluated in the context of the capillary and mitochondrial network. Results MPM revealed that 43.6 ± 3.3% of oxidative fiber capillaries had ≥ 50% of their circumference embedded in a groove in the sarcolemma, in vivo. Embedded capillaries were tightly associated with dense mitochondrial populations lateral to capillary grooves and nearly absent below the groove. Mitochondrial distribution, number of embedded capillaries, and capillary-to-fiber contact were proportional to fiber oxidative capacity and unaffected by myoglobin knockout. GLUT4 did not preferentially localize to embedded capillaries. Conclusions Embedding capillaries in the sarcolemma may provide a regulatory mechanism to optimize delivery of oxygen to heterogeneous groups of muscle fibers. We hypothesize that mitochondria locate to paravascular regions due to myofibril voids created by embedded capillaries, not to enhance the delivery of oxygen to the mitochondria. PMID:25279425

Fast monitoring of motor exhaust components by resonant multi-photon ionisation and time-of-flight mass spectrometry

Science.gov (United States)

Franzen, Jochen; Frey, Rüdiger; Nagel, Holger

1995-03-01

A new analytical procedure is provided by the combination of two types of spectroscopy. Resonant ionization of selected compounds by multiphoton ionization is based on results of absorption spectroscopy for the compound molecules of interest and time-of-flight mass spectrometry serves for the unambigious detection of these compounds. An interesting application of this method is the fast exhaust gas analysis. In the development of future combustion engines, the management of dynamic motor processes becomes predominant because by more than 90 % of all the dangerous exhaust pollutions are produced in instationary motor phases such as fast speed or load changes. The investigation of dynamic processes however, requires fast analytical procedures with millisecond time resolution together with the capability to measure individual components in a very complex gas mixture The objectives for a development project of such an instrument were set by the Research Association for Combustion Engines (Forschungsvereinigung Verbrennungskraftmaschinen, FVV, Germany): Up to ten substances should be monitored synchroneously with a time resolution of about 10 milliseconds, with concentration limits of 1 part per million and with a precision better than 10 % relative standard deviation. Such a laser mass spectrometer for fast multi-component automotive exhaust analyses has been developed in a joint research project by Bruker-Franzen Analytik GmbH, Dornier GmbH and the Technical University of Munich. The system has been applied at a motor test facility to investigate the emissions of the aromatic hydrocarbons benzene, toluene and xylene, of nitric oxide and acetaldehyde in stationary and dynamic engine operation. These measurements demonstrate that strong emission of these pollutants takes place at instationary engine operation and in particular that these compounds are emitted at different times, giving new information about the processes in the combustion chamber and in the exhaust pipe.

The X-ray microscopy project at saga SLS

International Nuclear Information System (INIS)

Yasumoto, M.; Ishiguro, E.; Takemoto, K.; Kihara, H.; Kamijo, N.; Tomimasu, T.; Tsurushima, T.; Takahara, A.; Hara, K.; Chikaura, Y.

2002-01-01

A new high resolution X-ray microscopy project has been proposed at Saga synchrotron light source, which is a third generation synchrotron light facility in Japan. Two microscopy beamlines are planned for this project. One is a scanning microscope in the water window region, and the other is a full-field imaging microscope in the multi-keV X-ray energy region. To demonstrate the feasibility of the project, the optical layout of the scanning microscope was designed. The beamline mainly consists of a 3.5 cm periodical undulator, a varied line-spacing plane grating monochromator (600 lines/mm) and an end-station including a zone plate. Thus, the calculated X-ray properties focused on the sample position are as follows: the spot size is ∼ 70 nm, the monochromaticity is ∼2000, and the photon flux is 10 9 ∼ 10 10 photons/sec. (authors)

Correlated Light Microscopy and Electron Microscopy

NARCIS (Netherlands)

Sjollema, Klaas A.; Schnell, Ulrike; Kuipers, Jeroen; Kalicharan, Ruby; Giepmans, Ben N. G.; MullerReichert, T; Verkade, P

2012-01-01

Understanding where, when, and how biomolecules (inter)act is crucial to uncover fundamental mechanisms in cell biology. Recent developments in fluorescence light microscopy (FLM) allow protein imaging in living cells and at the near molecular level. However, fluorescence microscopy only reveals

Best practices for managing large CryoEM facilities.

Science.gov (United States)

Alewijnse, Bart; Ashton, Alun W; Chambers, Melissa G; Chen, Songye; Cheng, Anchi; Ebrahim, Mark; Eng, Edward T; Hagen, Wim J H; Koster, Abraham J; López, Claudia S; Lukoyanova, Natalya; Ortega, Joaquin; Renault, Ludovic; Reyntjens, Steve; Rice, William J; Scapin, Giovanna; Schrijver, Raymond; Siebert, Alistair; Stagg, Scott M; Grum-Tokars, Valerie; Wright, Elizabeth R; Wu, Shenping; Yu, Zhiheng; Zhou, Z Hong; Carragher, Bridget; Potter, Clinton S

2017-09-01

This paper provides an overview of the discussion and presentations from the Workshop on the Management of Large CryoEM Facilities held at the New York Structural Biology Center, New York, NY on February 6-7, 2017. A major objective of the workshop was to discuss best practices for managing cryoEM facilities. The discussions were largely focused on supporting single-particle methods for cryoEM and topics included: user access, assessing projects, workflow, sample handling, microscopy, data management and processing, and user training. Crown Copyright © 2017. Published by Elsevier Inc. All rights reserved.

Facile hydrothermal synthesis of CeO 2 nanopebbles

Indian Academy of Sciences (India)

Cerium oxide (CeO2) nanopebbles have been synthesized using a facile hydrothermal method. X-ray diffraction pattern (XRD) and transmission electron microscopy analyses confirm the presence of CeO2 nanopebbles. XRD shows the formation of cubic fluorite CeO2 and the average particle size estimated from the ...

Microscope-Based Fluid Physics Experiments in the Fluids and Combustion Facility on ISS

Science.gov (United States)

Doherty, Michael P.; Motil, Susan M.; Snead, John H.; Malarik, Diane C.

2000-01-01

At the NASA Glenn Research Center, the Microgravity Science Program is planning to conduct a large number of experiments on the International Space Station in both the Fluid Physics and Combustion Science disciplines, and is developing flight experiment hardware for use within the International Space Station's Fluids and Combustion Facility. Four fluids physics experiments that require an optical microscope will be sequentially conducted within a subrack payload to the Fluids Integrated Rack of the Fluids and Combustion Facility called the Light Microscopy Module, which will provide the containment, changeout, and diagnostic capabilities to perform the experiments. The Light Microscopy Module is planned as a fully remotely controllable on-orbit microscope facility, allowing flexible scheduling and control of experiments within International Space Station resources. This paper will focus on the four microscope-based experiments, specifically, their objectives and the sample cell and instrument hardware to accommodate their requirements.

Order of multiphoton excitation of sulfonium photo-acid generators used in photoresists based on SU-8

Science.gov (United States)

Williams, Henry E.; Diaz, Carlos; Padilla, Gabriel; Hernandez, Florencio E.; Kuebler, Stephen M.

2017-06-01

Multiphoton lithography (MPL), Z-scan spectroscopy, and quantum chemical calculations were employed to investigate the order of multiphoton excitation that occurs when femtosecond laser pulses are used to excite two sulfonium photo-acid generators (PAGs) commonly used in photoresists based on the cross-linkable epoxide SU-8. The mole-fractions of the mono- and bis-sulfonium forms of these PAGs were determined for the commercially available photoresist SU-8 2075 and for the PAGs alone from a separate source. Both were found to contain similar fractions of the mono- and bis-forms, with the mono form present in the majority. Reichert's method was used to determine the solvatochromic strength of the SU-8 matrix, so that results obtained for the PAGs in SU-8 and in solution could be reliably compared. The PAGs were found to exhibit a minimal solvatochromic shift for a series of solvents that span across the solvatochromic strength of SU-8 itself. Sub-micron-sized features were fabricated in SU-8 2075 by MPL using amplified and continuous-wave mode-locked laser pulses. Analysis of the features as a function of average laser power, scan speed, and excitation wavelength shows that the PAGs can be activated by both two- and three-photon absorption (2PA and 3PA). Which activation mode dominates depends principally upon the excitation wavelength because the average laser powers that can be used with the photoresist are limited by practical considerations. The power must be high enough to effect sufficient cross-linking, yet not so high as to exceed the damage threshold of the material. When the laser pulses have a duration on the order of 100 fs, 3PA dominates at wavelengths near 800 nm, whereas 2PA becomes dominant at wavelengths below 700 nm. These findings are corroborated by open-aperture Z-scan measurements and quantum chemical calculations of the cross-sections for 2PA and 3PA as a function of wavelength.

Dictionary of Microscopy

Science.gov (United States)

Heath, Julian

2005-10-01

The past decade has seen huge advances in the application of microscopy in all areas of science. This welcome development in microscopy has been paralleled by an expansion of the vocabulary of technical terms used in microscopy: terms have been coined for new instruments and techniques and, as microscopes reach even higher resolution, the use of terms that relate to the optical and physical principles underpinning microscopy is now commonplace. The Dictionary of Microscopy was compiled to meet this challenge and provides concise definitions of over 2,500 terms used in the fields of light microscopy, electron microscopy, scanning probe microscopy, x-ray microscopy and related techniques. Written by Dr Julian P. Heath, Editor of Microscopy and Analysis, the dictionary is intended to provide easy navigation through the microscopy terminology and to be a first point of reference for definitions of new and established terms. The Dictionary of Microscopy is an essential, accessible resource for: students who are new to the field and are learning about microscopes equipment purchasers who want an explanation of the terms used in manufacturers' literature scientists who are considering using a new microscopical technique experienced microscopists as an aide mémoire or quick source of reference librarians, the press and marketing personnel who require definitions for technical reports.

Nanostructures based on quantum dots for application in promising methods of single- and multiphoton imaging and diagnostics

Science.gov (United States)

Nabiev, I. R.

2017-01-01

Molecules recognizing biomarkers of diseases (monoclonal antibodies (monoABs)) are often too large for biomedical applications, and the conditions that are used to bind them with nanolabels lead to disordered orientation of monoABs with respect to the nanoparticle surface. Extremely small nanoprobes, designed via oriented conjugation of quantum dots (QDs) with single-domain antibodies (sdABs) derived from the immunoglobulin of llama and produced in the E. coli culture, have a hydrodynamic diameter less than 12 nm and contain equally oriented sdAB molecules on the surface of each QD. These nanoprobes exhibit excellent specificity and sensitivity in quantitative determination of a small number of cells expressing biomarkers. In addition, the higher diffusion coefficient of sdABs makes it possible to perform immunohistochemical analysis in bulk tissue, inaccessible for conventional monoABs. The necessary conditions for implementing high-quality immunofluorescence diagnostics are a high specificity of labeling and clear differences between the fluorescence of nanoprobes and the autofluorescence of tissues. Multiphoton micros-copy with excitation in the near-IR spectral range, which is remote from the range of tissue autofluorescence excitation, makes it possible to solve this problem and image deep layers in biological tissues. The two-photon absorption cross sections of CdSe/ZnS QDs conjugated with sdABs exceed the corresponding values for organic fluorophores by several orders of magnitude. These nanoprobes provide clear discrimination between the regions of tumor and normal tissues with a ratio of the sdAB fluorescence to the tissue autofluorescence upon two-photon excitation exceeding that in the case of single-photon excitation by a factor of more than 40. The data obtained indicate that the sdAB-QD conjugates used as labels provide the same, or even better, quality as the "gold standard" of immunohistochemical diagnostics. The developed nanoprobes are expected to

Stanford MFEL and Near Infrared Science Center

Science.gov (United States)

2011-01-28

are incorporated into glass catadioptric lenses that are mounted and sealed at each end of the stainless steel microscope. In addition to the self...highly effective in preventing biofilm formation , as well as in killing biofilms that are already present. b) Peer-Reviewed publications (in reversed...Multiphoton Microscopy in the Biomedical Sciences VII, SPIE, vol. 6442 (2007). 3. On Image formation in Near-field Infrared Microscopy, D. M

High-resolution, 2- and 3-dimensional imaging of uncut, unembedded tissue biopsy samples.

Science.gov (United States)

Torres, Richard; Vesuna, Sam; Levene, Michael J

2014-03-01

Despite continuing advances in tissue processing automation, traditional embedding, cutting, and staining methods limit our ability for rapid, comprehensive visual examination. These limitations are particularly relevant to biopsies for which immediate therapeutic decisions are most necessary, faster feedback to the patient is desired, and preservation of tissue for ancillary studies is most important. The recent development of improved tissue clearing techniques has made it possible to consider use of multiphoton microscopy (MPM) tools in clinical settings, which could address difficulties of established methods. To demonstrate the potential of MPM of cleared tissue for the evaluation of unembedded and uncut pathology samples. Human prostate, liver, breast, and kidney specimens were fixed and dehydrated by using traditional histologic techniques, with or without incorporation of nucleic acid fluorescent stains into dehydration steps. A benzyl alcohol/benzyl benzoate clearing protocol was substituted for xylene. Multiphoton microscopy was performed on a home-built system. Excellent morphologic detail was achievable with MPM at depths greater than 500 μm. Pseudocoloring produced images analogous to hematoxylin-eosin-stained images. Concurrent second-harmonic generation detection allowed mapping of collagen. Subsequent traditional section staining with hematoxylin-eosin did not reveal any detrimental morphologic effects. Sample immunostains on renal tissue showed preservation of normal reactivity. Complete reconstructions of 1-mm cubic samples elucidated 3-dimensional architectural organization. Multiphoton microscopy on cleared, unembedded, uncut biopsy specimens shows potential as a practical clinical tool with significant advantages over traditional histology while maintaining compatibility with gold standard techniques. Further investigation to address remaining implementation barriers is warranted.

Heavy-ion microscopy

International Nuclear Information System (INIS)

Kraft, G.; Yang, T.C.H.; Richards, T.; Tobias, C.A.

1980-01-01

This chapter briefly describes the techniques of optical microscopy, scanning and transmission electron microscopy, soft x-ray microscopy and compares these latter techniques with heavy-ion microscopy. The resolution obtained with these various types of microscopy are compared and the influence of the etching procedure on total resolution is discussed. Several micrographs of mammalian cells are included

Location-dependent coronary artery diffusive and convective mass transport properties of a lipophilic drug surrogate measured using nonlinear microscopy.

Science.gov (United States)

Keyes, Joseph T; Simon, Bruce R; Vande Geest, Jonathan P

2013-04-01

Arterial wall mass transport properties dictate local distribution of biomolecules or locally delivered dugs. Knowing how these properties vary between coronary artery locations could provide insight into how therapy efficacy is altered between arterial locations. We introduced an indocarbocyanine drug surrogate to the lumens of left anterior descending and right coronary (LADC; RC) arteries from pigs with or without a pressure gradient. Interstitial fluorescent intensity was measured on live samples with multiphoton microscopy. We also measured binding to porcine coronary SMCs in monoculture. Diffusive transport constants peaked in the middle sections of the LADC and RC arteries by 2.09 and 2.04 times, respectively, compared to the proximal and distal segments. There was no statistical difference between the average diffusivity value between LADC and RC arteries. The convection coefficients had an upward trend down each artery, with the RC being higher than the LADC by 3.89 times. This study demonstrates that the convective and diffusive transport of lipophilic molecules changes between the LADC and the RC arteries as well as along their length. These results may have important implications in optimizing drug delivery for the treatment of coronary artery disease.

Managing the Introduction of Super-Resolution Microscopy into a Core Facility.

Science.gov (United States)

Kamykowski, Jeffrey A; Storrie, Brian

2017-01-01

Super resolution techniques place the resolution of fluorescence microscopy closer to the size of the underlying cell structure or molecular machine being studied. Structured illumination techniques will give users a set of tools that are close to their past experience and relatively simple and quick to learn. The present dyes can be used. Resolution approaching 100 nm XY can be achieved. In contrast, stochastic methods such as PALM/STORM typically require the choice of new dyes and a much greater learning curve to master the technology and calculations. However, a further fivefold resolution improvement is possible. Stimulated depletion techniques such as STED offer a third set of approaches that will again require the use of new dyes. All these approaches require substantial investment in new equipment and in user training. There is no free lunch in the search for better resolution.

Theory of the multiphoton cascade transitions with two photon links: comparison of quantum electrodynamical and quantum mechanical approaches

International Nuclear Information System (INIS)

Zalialiutdinov, T; Baukina, Yu; Solovyev, D; Labzowsky, L

2014-01-01

The theory of multiphoton cascade transitions with two-photon links is considered within two different approaches: quantum electrodynamical (QED) and phenomenological quantum mechanical (QM). A problem of regularization of the cascade contributions is investigated in detail. It is argued that the correct regularization should include both initial and intermediate level widths in the singular energy denominators. This result follows both from the QED and from the QM approach. Particular transitions nl → 1s + 2γ with nl = 3s, 4s, 3d, 4d and nl → 1s + 3γ with nl = 3p, 4p are considered as examples. The importance of the proper cascade regularization is also demonstrated. (paper)

Enhancements to the analytical facilities at the GNS proton microprobe

International Nuclear Information System (INIS)

Barry, B.J.; Markwitz, A.; Kennedy, V.J.; Trompetter, W.J.

2005-01-01

In recent years a number of detection systems have been added to the proton microprobe facility at GNS Science. Particular additions have been a large area HPGe detector and a system for scanning transmission imaging microscopy. The HPGe detector has improved detection sensitivity, particularly for higher energy K x-rays where energy resolution is of lesser importance. The scanning microscopy system has enabled mapping of areal densities in biological samples to give accurate elemental maps. Examples are given of these applications. (author). 22 refs., 7 figs

Building collaboration tools and access to on-line facilities

International Nuclear Information System (INIS)

Agarwal, D.; Sachs, S.

1996-11-01

Network-based facilities will allow researchers at different locations to collaborate on experiments as if they all were together in the same laboratory. The expected value of these geographically distributed environments includes substantially increased effectiveness in doing science, and an enabling capability for analytical and high-value production use by industry. The Distributed, Collaboratory Experiment Environments (DCEE) Program consists of four projects that were established to build prototype remote experiment and collaborative environments. The work undertaken in this project represents some of the research and development of the mechanisms and infrastructure required to make collaboratories a reality. Some of these mechanisms have already been developed. Several other mechanisms, such as data dissemination, resource management for the sharing of experiment control, safety and security, electronic notebooks, elements of telepresence, and integrated user interfaces need further research and development. The pilot application for these collaborative tools is the Advanced Light Source (ALS) Beamline 7.0 at the Ernest Orlando Lawrence Berkeley Laboratory. The ALS is a particle accelerator and is a source of very high brilliance soft X-ray beams. One experimental facility is the Spectro-Microscopy Facility Beamline 7.0. Through this project, the Spectro-Microscopy Facility will be opened up to users from a wide range of organizations. The goal is to build software that will not only put the ALS Beamline 7.0 on-line, but will also serve as building blocks for future collaboratory development

Population inversion of two atoms under the phase decoherence in the multiphoton process

International Nuclear Information System (INIS)

Zhang Dongxia; Sa Chuerfu; Mu Qier

2011-01-01

By means of the quantum theory, the population inversion of two atoms in the system of two two-level atoms coupled to a light field in the Binomial Optical Field are investigated in the presence of phase decoherence in the multiphoton Tavis-Cumming Model. The influences of the phase decoherence coefficient, the parameters η of the binomial optical field, the maximum number of photons and the number of the transitional photons on the properties of the population inversion of two atoms have been discussed. The results show that the phase decoherence reduced the oscillation amplitude of the population inversion of two atoms and destroyed the atomic quantum characteristic. Changing the number of the transitional photons, evolved cycle and evolved intensity the population inversion of two atoms can be changed. The phenomena of collapse and revival disappear as photon number increase. When the binomial optical state changes from a coherent state to a Fock state, the oscillation frequency of the atomic population reduces gradually, the phenomena of collapse and revival vanishes gradually. (authors)

Innovative Strategies for Clinical Microscopy Instruction: Virtual Versus Light Microscopy.

Science.gov (United States)

McDaniel, M Jane; Russell, Gregory B; Crandall, Sonia J

2018-06-01

The purpose of the study was to compare virtual microscopy with light microscopy to determine differences in learning outcomes and learner attitudes in teaching clinical microscopy to physician assistant (PA) students. A prospective, randomized, crossover design study was conducted with a convenience sample of 67 first-year PA students randomized to 2 groups. One group used light microscopes to find microscopic structures, whereas the other group used instructor-directed video streaming of microscopic elements. At the midpoint of the study, the groups switched instructional strategies. Learning outcomes were assessed via posttest after each section of the study, with comparison of final practical examination results to previous cohorts. Attitudes about the 2 educational strategies were assessed through a postcourse questionnaire with a Likert scale. Analysis of the first posttest demonstrated that students in the video-streamed group had significantly better learning outcomes than those in the light microscopy group (P = .004; Cohen's d = 0.74). Analysis of the posttest after crossover showed no differences between the 2 groups (P = .48). Between the 2 posttests, students first assigned to the light microscopy group scored a 6.6 mean point increase (±10.4 SD; p = .0011), whereas students first assigned to the virtual microscopy group scored a 1.3 mean point increase (±7.1 SD; p = .29). The light microscopy group improved more than the virtual microscopy group (P = .019). Analysis of practical examination data revealed higher scores for the study group compared with 5 previous cohorts of first-year students (P virtual microscopy to traditional light microscopy. Virtual microscopy is an effective educational strategy, and students prefer this method when learning to interpret images of clinical specimens.

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

Science.gov (United States)

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

2010-02-26

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

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

International Nuclear Information System (INIS)

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

2010-01-01

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

Compression of fiber supercontinuum pulses to the Fourier-limit in a high-numerical-aperture focus

DEFF Research Database (Denmark)

Tu, Haohua; Liu, Yuan; Turchinovich, Dmitry

2011-01-01

A multiphoton intrapulse interference phase scan (MIIPS) adaptively and automatically compensates the combined phase distortion from a fiber supercontinuum source, a spatial light modulator pulse shaper, and a high-NA microscope objective, allowing Fourier-transform-limited compression of the sup......A multiphoton intrapulse interference phase scan (MIIPS) adaptively and automatically compensates the combined phase distortion from a fiber supercontinuum source, a spatial light modulator pulse shaper, and a high-NA microscope objective, allowing Fourier-transform-limited compression...... power of 18–70mW, and a repetition rate of 76MHz, permitting the application of this source to nonlinear optical microscopy and coherently controlled microspectroscopy....

Comparative study of Nd(3+) emission from 4f2 5d and 4f3 configurations induced by multiphotonic process in YLF, GLF and LLF crystals

International Nuclear Information System (INIS)

Librantz, Andre Felipe Henriques

2000-01-01

Nd 3+ ultraviolet fluorescence induced by multiphotonic laser excitations was studied in Nd-doped YLiF 4 (YLF) and LuLiF 4 (LLF) crystals by using the time resolved spectroscopy technique. The UV luminescences are due to transitions between the 4f 2 5d and the 4f 3 electronic configurations of Nd 3+ ions. The 4f 2 5d configuration can be reached by direct pumping the UV transition or by multiphotonic excitation, both processes give raise to the UV emission band with a structure due to the strong phonon coupling, expected for a 5d orbital involvement in the transition. The multiphotonic excitation process is due to three photons (532 nm) sequential absorptions of 532 nm-photons by metastable levels of the 4f 3 configuration splitted by crystalline local field. The sequential excitation of Nd by the pumping laser is attributed to the 4 I 9/2 +532nm → 4 G 7/2 ground state absorption followed by the 4 G 7/2 +532 nm →2 F 5/2 and 2 F 5/2 +532 nm → 4f 2 5d excited state absorptions. The UV emissions due to 4f 2 5d configuration are parity allowed, having lifetime of 35 ns in contrast to UV emissions from 4f 3 configuration which are induced by two absorption steps and are parity forbidden showing longer lifetime of 8μs and narrow tines. The polarization effects of the UV emissions were studied and their behavior are dependent on the excited state configuration involving or not the 5d orbital. The allowed UV emissions positions were affected by the host variation more than the ones originating from the 4f 3 configuration as expected. The electronic energy of the 4f 2 5d configuration shifts to lower energy when increasing the crystal field. (author)

Environmental Microscopy Facility

Data.gov (United States)

Federal Laboratory Consortium — FUNCTION: Provides high-resolution (5 nm) images and elemental composition (for elements heavier than sodium) of hydrated specimens, including biological materials....

Rapid creation of distant entanglement by multi-photon resonant fluorescence

Science.gov (United States)

Cohen, Guy Z.; Sham, L. J.

2014-03-01

We study a simple, effective and robust method for entangling two separate stationary quantum dot spin qubits with high fidelity using multi-photon Gaussian state. The fluorescence signals from the two dots interfere at a beam splitter. The bosonic nature of photons leads, in analogy with the Hong-Ou-Mandel (HOM) effect, to selective pairing of photon holes (photon absences in the fluorescent signals). By the HOM effect, two photon holes with the same polarization end up at the same beam splitter output. As a result, two odd photon number detections at the outgoing beams, which must correspond to two photon holes with different polarizations, herald entanglement creation. The robustness of the Gaussian states is evidenced by the ability to compensate for photon absorption and noise by a moderate increase in the number of photons at the input. We calculate the entanglement generation rate in the ideal, non-ideal and near-ideal detector regimes and find substantial improvement over single-photon schemes in all three regimes. Fast and efficient spin-spin entanglement creation can form the basis for a scalable quantum dot quantum computing network. Our predictions can be tested using current experimental capabilities. This research was supported by the U.S. Army Research Office MURI award W911NF0910406, by NSF grant PHY-1104446 and by ARO (IARPA, W911NF-08-1-0487). The authors thank D. G. Steel for useful discussions.

Status of FEL-SUT project, and the experimental setup for multiphoton dissociation and isotope separation in the gaseous phase

CERN Document Server

Chernyshev, A V; Petrov, A K; Kawai, M; Toyoda, K; Nakai, K; Kuroda, H

2001-01-01

The IR FEL Research Center of the Science University of Tokyo (FEL-SUT) is open for users to develop new applications of IR FEL in a wide field of material science, chemical technology and bio-chemical applications. The FEL is based on 35 MeV linac operated at the frequency of 2856 MHz (s-band). The FEL covers the wavelength range from 5 to 16 mu m with the micropulse duration of 1-2 ps, macropulse duration of 1 mu s, macropulse repetition rate of 10 Hz and the overall average power of 1 W. We report the present status of the Center and an experimental setup designed and constructed for the experiments on multiphoton dissociation and isotope separation.

In situ ion irradiation/implantation studies in the HVEM-Tandem Facility at Argonne National Laboratory

International Nuclear Information System (INIS)

Allen, C.W.; Funk, L.L.; Ryan, E.A.; Taylor, A.

1988-09-01

The HVEM-Tandem User Facility at Argonne National Laboratory interfaces two ion accelerators, a 2 MV tandem accelerator and a 650 kV ion implanter, to a 1.2 MV high voltage electron microscope. This combination allows experiments involving simultaneous ion irradiation/ion implantation, electron irradiation and electron microscopy/electron diffraction to be performed. In addition the availability of a variety of microscope sample holders permits these as well as other types of in situ experiments to be performed at temperatures ranging from 10-1300 K, with the sample in a stressed state or with simultaneous determination of electrical resistivity of the specimen. This paper summarizes the details of the Facility which are relevant to simultaneous ion beam material modification and electron microscopy, presents several current applications and briefly describes the straightforward mechanism for potential users to access this US Department of Energy supported facility. 7 refs., 1 fig., 1 tab

Progress in the Correlative Atomic Force Microscopy and Optical Microscopy

Directory of Open Access Journals (Sweden)

Lulu Zhou

2017-04-01

Full Text Available Atomic force microscopy (AFM has evolved from the originally morphological imaging technique to a powerful and multifunctional technique for manipulating and detecting the interactions between molecules at nanometer resolution. However, AFM cannot provide the precise information of synchronized molecular groups and has many shortcomings in the aspects of determining the mechanism of the interactions and the elaborate structure due to the limitations of the technology, itself, such as non-specificity and low imaging speed. To overcome the technical limitations, it is necessary to combine AFM with other complementary techniques, such as fluorescence microscopy. The combination of several complementary techniques in one instrument has increasingly become a vital approach to investigate the details of the interactions among molecules and molecular dynamics. In this review, we reported the principles of AFM and optical microscopy, such as confocal microscopy and single-molecule localization microscopy, and focused on the development and use of correlative AFM and optical microscopy.

Design and installation of a multimode microscopy system

Science.gov (United States)

Helm, Johannes P.; Haug, Finn-Mogens S.; Storm, Johan F.; Ottersen, Ole-Petter

2001-04-01

We describe design and installation of a multi-mode microscopy core facility in an environment of varied research activity in life-sciences. The experimentators can select any combination of a) microscopes (upright, upright fixed-stage, inverted), b) microscopy modes (widefield, DIC, IRDIC, widefield epifluorescence, transmission LSM, reflection and fluorescence CLSM, MPLSM), c) imaging techniques (direct observation, video observation, photography, quantitative camera-recording, flying spot scanning), d) auxiliary systems (equipment for live specimen imaging, electrophysiology, time-coordinated laser-scanning and electrophysiology, patch-clamp). The equipment is installed on one large vibration-isolating optical table (3m X 1.5m X 0.3m). Electronics, auxiliary equipment, and a fiber-coupled, remotely controlled Ar+-Kr+ laser are mounted in a rack system fixed to the ceiling. The design of the shelves allows the head of the CSLM to be moved to any of the microscopes without increasing critical cable lengths. At the same time easy access to all the units is preserved. The beam of a Titanium-Sapphire laser, controlled by means of an EOM and a prism GVD, is coupled directly to the microscopes. Three mirrors mounted on a single precision translation table are integrated into the beam steering system so that the beam can easily be redirected to any of the microscopes. All the available instruments can be operated by the educated and trained user. The system is popular among researchers in neuroanatomy, embryology, cell biology, molecular biology - including the study of protein interactions, e.g. by means of FRET, and electrophysiology. Its colocalization with an EM facility promises to provide considerable synergy effects.

Cellular organization and spectral diversity of GFP-like proteins in live coral cells studied by single and multiphoton imaging and microspectroscopy

Science.gov (United States)

Salih, Anya; Cox, Guy C.; Larkum, Anthony W.

2003-07-01

Tissues of many marine invertebrates of class Anthozoa contain intensely fluorescent or brightly coloured pigments. These pigments belong to a family of photoactive proteins closely related to Green Fluorescent Protein (GFP), and their emissions range from blue to red wavelengths. The great diversity of these pigments has only recently been realised. To investigate the role of these proteins in corals, we have performed an in vivo fluorescent pigment (FP) spectral and cellular distribution analyses in live coral cells using single and multi-photon laser scanning imaging and microspectroscopy. These analyses revealed that even single colour corals contain spectroscopically heterogeneous pigment mixtures, with 2-5 major colour types in the same area of tissue. They were typically arranged in step-wise light emission energy gradients (e.g. blue, green, yellow, red). The successive overlapping emission-excitation spectral profiles of differently coloured FPs suggested that they were suited for sequential energy coupling. Traces of red FPs (emission = 570-660 nm) were present, even in non-red corals. We confirmed that radiative energy transfer could occur between separate granules of blue and green FPs and that energy transfer was inversely proportional to the square of the distance between them. Multi-photon micro-spectrofluorometric analysis gave significantly improved spectral resolution by restricting FP excitation to a single point in the focal plane of the sample. Pigment heterogeneity at small scales within granules suggested that fluorescence resonance energy transfer (FRET) might be occurring, and we confirmed that this was the case. Thus, energy transfer can take place both radiatively and by FRET, probably functioning in photoprotection by dissipation of excessive solar radiation.

Pico-second laser materials interactions: mechanisms, material lifetime and performance optimization Ted Laurence(14-ERD-014)

Energy Technology Data Exchange (ETDEWEB)

Laurence, Ted A. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

2016-12-14

Laser-induced damage with ps pulse widths straddles the transition from intrinsic, multiphoton ionization- and avalanche ionization-based ablation with fs pulses to defectdominated, thermal-based damage with ns pulses. We investigated the morphology and scaling of damage for commonly used silica and hafnia coatings as well as fused silica. Using carefully calibrated laser-induced damage experiments, in situ imaging, and high-resolution optical microscopy, atomic force microscopy, and scanning electron microscopy, we showed that defects play an important role in laser-induced damage for pulse durations as short as 1 ps. Three damage morphologies were observed: standard material ablation, ultra-high density pits, and isolated absorbers. For 10 ps and longer, the isolated absorbers limited the damage performance of the coating materials. We showed that damage resulting from the isolated absorbers grows dramatically with subsequent pulses for sufficient fluences. For hafnia coatings, we used electric field modeling and experiments to show that isolated absorbers near the surface were affected by the chemical environment (vacuum vs. air) for pulses as short as 10 ps. Coupled with the silica results, these results suggested that improvements in the performance in the 10 -60 ps range have not reached fundamental limits. These findings motivate new efforts, including a new SI LDRD in improving the laser-damage performance of multi-layer dielectric coatings. A damage test facility for ps pulses was developed and automated, and was used for testing production optics for ARC. The resulting software was transferred to other laser test facilities for fs pulses and multiple wavelengths with 30 ps pulses. Additionally, the LDRD supported the retention and promotion of an important staff scientist in high-resolution dynamic microscopy and laser-damage testing.

Electron Microscopy Facility for Research and Services in the Malaysian Nuclear Agency towards TSO

International Nuclear Information System (INIS)

Nadira Kamarudin; Mohd Bin Harun; Zaiton Selamat

2011-01-01

Scanning Electron Microscope FEI-Quanta 400 (SEM) made in the USA was commissioned in late 2003. This equipment is used in many areas of materials science, metallurgy, engineering, electronics, medicine, agriculture, biology and so on. This facility has helped the researchers in conducting research in their respective fields as well have been providing services to agencies, institutions, industries and local industry. Since 2004, there were 81 projects and 5000 samples analyzed using this facility in Malaysian Nuclear Agency, while 23 companies and 900 samples were from various agencies. In addition, revenue derived from these services has able to provide for the maintenance of this equipment. SEM is an important step in the nuclear material testing process. Nuclear material can be inspected for its performance by getting information from its morphology micrograph by using SEM. It opens up a whole new world that is unseen by the naked eye. (author)

French Society of Microscopy, 10. conference; Societe Francaise des Microscopies, 10. colloque

Energy Technology Data Exchange (ETDEWEB)

Thibault-Penisson, J; Cremer, Ch; Susini, J; Kirklanda, A I; Rigneault, H; Renault, O; Bailly, A; Zagonel, L F; Barrett, N; Bogner, A; Gauthier, C; Jouneau, P H; Thollet, G; Fuchs, G; Basset, D; Deconihout, B; Vurpillot, F; Vella, A; Matthieu, G; Cadel, E; Bostel, A; Blavette, D; Baumeister, W; Usson, Y; Zaefferer, St; Laffont, L; Weyland, M; Thomas, J M; Midgley, P; Benlekbir, S; Epicier, Th; Diop, B N; Roux, St; Ou, M; Perriat, P; Bausach, M; Aouine, M; Berhault, G; Idrissi, H; Cottevieille, M; Jonic, S; Larquet, E; Svergun, D; Vannoni, M A; Boisset, N; Ersena, O; Werckmann, J; Ulhaq, C; Hirlimann, Ch; Tihay, F; Cuong, Pham-Huu; Crucifix, C; Schultz, P; Jornsanoha, P; Thollet, G; Masenelli-Varlot, K; Gauthier, C; Ludwig, W; King, A; Johnson, G; Gonzalves-Hoennicke, M; Reischig, P; Messaoudi, C; Ibrahim, R; Marco, S; Klie, R F; Zhao, Y; Yang, G; Zhu, Y; Hue, F; Hytch, M; Hartmann, J M; Bogumilowicz, Y; Claverie, A; Klein, H; Alloyeau, D; Ricolleau, C; Langlois, C; Le Bouar, Y; Loiseau, A; Colliex, C; Stephan, O; Kociak, M; Tence, M; Gloter, A; Imhoff, D; Walls, M; Nelayah, J; March, K; Couillard, M; Ailliot, C; Bertin, F; Cooper, D; Rivallin, P; Dumelie, N; Benhayoune, H; Balossier, G; Cheynet, M; Pokrant, S; Tichelaar, F; Rouviere, J L; Cooper, D; Truche, R; Chabli, A; Debili, M Y; Houdellier, F; Warot-Fonrose, B; Hytch, M J; Snoeck, E; Calmels, L; Serin, V; Schattschneider, P; Jacob, D; Cordier, P

2007-07-01

This document gathers the resumes of some of the presentations made at this conference whose aim was to present the last developments and achievements of the 3 complementary microscopies: optical microscopy, electron microscopy and X-ray microscopy. The contributions have been organized around the following 12 topics: 1) new technical developments, 2) 3-dimensional imaging, 3) quantitative microscopy, 4) technical progress in photon microscopy, 5) synchrotron radiation, 6) measurements of patterns, deformations and strains, 7) materials for energy and transports, 8) nano-structures, 9) virus: structure and infection mechanisms, 10) 3-dimensional imaging for molecules, cells and cellular tissues, 11) nano-particles and colloids, and 12) liquid crystals.

Metabolic Mapping of Breast Cancer with Multiphoton Spectral and Lifetime Imaging

Science.gov (United States)

2007-03-01

2002. Spectrally resolved fluorescence lifetime imaging microscopy. Appl. Spec- trosc. 56 :155-166. 38. Becker, W., A. Bergmann, E. Haustein , Z...photon fluores- cence lifetime imaging microscopy of macrophage-mediated antigen processing. J. Microsc. 185 :339-353. 45. Lin, H.J., P. Herman , and

On the necessity of taking into account the contribution of multiphoton exchanges into electron-proton deep inelastic scattering

International Nuclear Information System (INIS)

Savrin, V.I.

1979-01-01

The hypothesis that the multiphoton exchanges give a substantial contribution to the electron-proton inclusive scattering is formulated. The hypothesis explains the observed violation of the Bjorken scaling law. As it is shown, the mechanism of such intensification of multiple exchanges may by connected with the properties of the processes of hadron multiproduction in the deep inelastic field. This results in the necessity to calculate the inclusive cross section in all electromagnetic coupling constant orders. This has been done in the framework of the density matrix method. As a result the deep inelastic scattering cross section calculated without application of the perturbation theory reveals a new property of the scaling invariance and leads to the natural relationship of structural functions with electromagnetic proton form-factors on the exclusive threshold

Ultra slow muon microscopy by laser resonant ionization at J-PARC, MUSE

Science.gov (United States)

Miyake, Y.; Ikedo, Y.; Shimomura, K.; Strasser, P.; Kawamura, N.; Nishiyama, K.; Koda, A.; Fujimori, H.; Makimura, S.; Nakamura, J.; Nagatomo, T.; Kadono, R.; Torikai, E.; Iwasaki, M.; Wada, S.; Saito, N.; Okamura, K.; Yokoyama, K.; Ito, T.; Higemoto, W.

2013-04-01

As one of the principal muon beam line at the J-PARC muon facility (MUSE), we are now constructing a Muon beam line (U-Line), which consists of a large acceptance solenoid made of mineral insulation cables (MIC), a superconducting curved transport solenoid and superconducting axial focusing magnets. There, we can extract 2 × 108/s surface muons towards a hot tungsten target. At the U-Line, we are now establishing a new type of muon microscopy; a new technique with use of the intense ultra-slow muon source generated by resonant ionization of thermal Muonium (designated as Mu; consisting of a μ + and an e - ) atoms generated from the surface of the tungsten target. In this contribution, the latest status of the Ultra Slow Muon Microscopy project, fully funded, is reported.

Coherent light microscopy

CERN Document Server

Ferraro, Pietro; Zalevsky, Zeev

2011-01-01

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

A series of fluorene-based two-photon absorbing molecules: synthesis, linear and nonlinear characterization, and bioimaging

Science.gov (United States)

Andrade, Carolina D.; Yanez, Ciceron O.; Rodriguez, Luis; Belfield, Kevin D.

2010-01-01

The synthesis, structural, and photophysical characterization of a series of new fluorescent donor–acceptor and acceptor-acceptor molecules, based on the fluorenyl ring system, with two-photon absorbing properties is described. These new compounds exhibited large Stokes shifts, high fluorescent quantum yields, and, significantly, high two-photon absorption cross sections, making them well suited for two-photon fluorescence microscopy (2PFM) imaging. Confocal and two-photon fluorescence microscopy imaging of COS-7 and HCT 116 cells incubated with probe I showed endosomal selectivity, demonstrating the potential of this class of fluorescent probes in multiphoton fluorescence microscopy. PMID:20481596

O2 rotational temperature measurements in an atmospheric air microdischarge by radar resonance-enhanced multiphoton ionization

International Nuclear Information System (INIS)

Sawyer, Jordan; Wu, Yue; Zhang, Zhili; Adams, Steven F.

2013-01-01

Nonintrusive spatially resolved rotational temperature measurements in an atmospheric air microdischarge are presented. The measurements were based on coherent microwave Rayleigh scattering (Radar) from resonance-enhanced multiphoton ionization of molecular oxygen. The open air DC microdischarge source operated in a stable “normal-glow” mode and pin-to-pin electrodes spaced 1.3 mm apart. The second harmonic of a tunable dye laser beam was focused between the two electrodes and scanned between 286 and 288 nm. Coherent microwave Rayleigh scattering was used to collect the two-photon rotational spectra of O 2 at C 3 Π(v = 2)←X 3 Σ(v′ = 0) transitions. The Boltzmann plots from analyses of the O 2 rotational lines determined local rotational temperatures at various axial locations between the electrodes. The molecular oxygen rotational temperature varied from ∼1150 K to ∼1350 K within the discharge area. The measurements had an accuracy of ∼±50 K.

O2 rotational temperature measurements in an atmospheric air microdischarge by radar resonance-enhanced multiphoton ionization

Science.gov (United States)

Sawyer, Jordan; Wu, Yue; Zhang, Zhili; Adams, Steven F.

2013-06-01

Nonintrusive spatially resolved rotational temperature measurements in an atmospheric air microdischarge are presented. The measurements were based on coherent microwave Rayleigh scattering (Radar) from resonance-enhanced multiphoton ionization of molecular oxygen. The open air DC microdischarge source operated in a stable "normal-glow" mode and pin-to-pin electrodes spaced 1.3 mm apart. The second harmonic of a tunable dye laser beam was focused between the two electrodes and scanned between 286 and 288 nm. Coherent microwave Rayleigh scattering was used to collect the two-photon rotational spectra of O2 at C3Π(v = 2)←X3Σ(v' = 0) transitions. The Boltzmann plots from analyses of the O2 rotational lines determined local rotational temperatures at various axial locations between the electrodes. The molecular oxygen rotational temperature varied from ˜1150 K to ˜1350 K within the discharge area. The measurements had an accuracy of ˜±50 K.

Bio-optic signatures for advanced glycation end products in the skin in streptozotocin (STZ) Induced Diabetes (Conference Presentation)

Science.gov (United States)

Saidian, Mayer; Ponticorvo, Adrien; Rowland, Rebecca A.; Balbado, Melisa L.; Lentsch, Griffin; Balu, Mihaela; Alexander, Micheal; Shiri, Li; Lakey, Jonathan R. T.; Durkin, Anthony J.; Kohen, Roni; Tromberg, Bruce J.

2017-02-01

Type 1diabetes (T1D) is an autoimmune disorder that occurs due to the rapid destruction of insulin-producing beta cells, leading to insulin deficiency and the inability to regulate blood glucose levels and leads to destructive secondary complications. Advanced glycation end (AGEs) products, the result of the cross-linking of reducing sugars and proteins within the tissues, are one of the key causes of major complications associated with diabetes such as renal failure, blindness, nerve damage and vascular changes. Non-invasive techniques to detect AGEs are important for preventing the harmful effects of AGEs during diabetes mellitus. In this study, we utilized multiphoton microscopy to image biopsies taken from control rats and compared them to biopsies taken from streptozotocin (STZ) induced adult male diabetic rats. This was done at two and four weeks after the induction of hyperglycemia (>400 mg/dL) specifically to evaluate the effects of glycation on collagen. We chose to use an in-situ multiphoton microscopy method that combines multiphoton auto-florescence (AF) and second harmonic generation (SHG) to detect the microscopic influence of glycation. Initial results show high auto-florescence levels were present on the collagen, as a result of the accumulation of AGEs only two weeks after the STZ injection and considerably higher levels were present four weeks after the STZ injection. Future projects could involve evaluating advanced glycation end products in a clinical trial of diabetic patients.

Multiphoton ionization in superintense, high-frequency laser fields. I. General developments

International Nuclear Information System (INIS)

Pont, M.

1991-01-01

This is the first of two papers studying multiphoton ionization (MPI) in superintense, high-frequency laser fields. They are based on a general iteration scheme in increasing powers of the inverse frequency. To lowest order in the frequency, i.e., the high-frequency limit, the atom was shown to be stable against decay by MPI, though distorted. To next order in the iteration, an expression for the MPI amplitude was obtained. In the present paper, we present general developments from this expression, valid for arbitrary polarization, binding potential, intensity, and initial state. First we analyze the symmetry of the angular distributions of photoelectrons determined by this expression for the MPI amplitude. This expression can explain the asymmetries in the angular distributions of photoelectrons occurring in the case of elliptic polarization that were recently reported in experiments. In the radiation regime where our theory applies these asymmetries are, however, weak. In certain instances our theory yields asymmetries in cases where lowest-order perturbation theory (LOPT) fails to predict them. We prove that at low intensities our expression for the MPI amplitude yields results in agreement with LOPT evaluated at high frequencies. An important part of this paper consists, however, of the derivation of an alternative form for the MPI amplitude of atomic hydrogen, which is substantially simpler, though somewhat less accurate. We study the consequences of this simplified expression for the case of linearly polarized fields in the following paper [Phys. Rev. A 44, xxxx (1991)

Imaging of Caenorhabditis elegans samples and sub-cellular localization of new generation photosensitizers for photodynamic therapy, using non-linear microscopy

Energy Technology Data Exchange (ETDEWEB)

Filippidis, G [Institute of Electronic Structure and Laser, Foundation of Research and Technology-Hellas, PO Box 1527, 71110 Heraklion (Greece); Kouloumentas, C [Institute of Electronic Structure and Laser, Foundation of Research and Technology-Hellas, PO Box 1527, 71110 Heraklion (Greece); Kapsokalyvas, D [Institute of Electronic Structure and Laser, Foundation of Research and Technology-Hellas, PO Box 1527, 71110 Heraklion (Greece); Voglis, G [Institute of Molecular Biology and Biotechnology, Foundation of Research and Technology, Heraklion 71110, Crete (Greece); Tavernarakis, N [Institute of Molecular Biology and Biotechnology, Foundation of Research and Technology, Heraklion 71110, Crete (Greece); Papazoglou, T G [Institute of Electronic Structure and Laser, Foundation of Research and Technology-Hellas, PO Box 1527, 71110 Heraklion (Greece)

2005-08-07

Two-photon excitation fluorescence (TPEF) and second-harmonic generation (SHG) are relatively new promising tools for the imaging and mapping of biological structures and processes at the microscopic level. The combination of the two image-contrast modes in a single instrument can provide unique and complementary information concerning the structure and the function of tissues and individual cells. The extended application of this novel, innovative technique by the biological community is limited due to the high price of commercial multiphoton microscopes. In this study, a compact, inexpensive and reliable setup utilizing femtosecond pulses for excitation was developed for the TPEF and SHG imaging of biological samples. Specific cell types of the nematode Caenorhabditis elegans were imaged. Detection of the endogenous structural proteins of the worm, which are responsible for observation of SHG signals, was achieved. Additionally, the binding of different photosensitizers in the HL-60 cell line was investigated, using non-linear microscopy. The sub-cellular localization of photosensitizers of a new generation, very promising for photodynamic therapy (PDT) (Hypericum perforatum L. extracts) was achieved. The sub-cellular localization of these novel photosensitizers was linked with their photodynamic action during PDT, and the possible mechanisms for cell killing have been elucidated.

Ion, X-ray, UV and Neutron Microbeam Systems for Cell Irradiation.

Science.gov (United States)

Bigelow, A W; Randers-Pehrson, G; Garty, G; Geard, C R; Xu, Y; Harken, A D; Johnson, G W; Brenner, D J

2010-08-08

The array of microbeam cell-irradiation systems, available to users at the Radiological Research Accelerator Facility (RARAF), Center for Radiological Research, Columbia University, is expanding. The HVE 5MV Singletron particle accelerator at the facility provides particles to two focused ion microbeam lines: the sub-micron microbeam II and the permanent magnetic microbeam (PMM). Both the electrostatic quadrupole lenses on the microbeam II system and the magnetic quadrupole lenses on the PMM system are arranged as compound lenses consisting of two quadrupole triplets with "Russian" symmetry. Also, the RARAF accelerator is a source for a proton-induced x-ray microbeam (undergoing testing) and is projected to supply protons to a neutron microbeam based on the (7)Li(p, n)(7)Be nuclear reaction (under development). Leveraging from the multiphoton microscope technology integrated within the microbeam II endstation, a UV microspot irradiator - based on multiphoton excitation - is available for facility users. Highlights from radiation-biology demonstrations on single living mammalian cells are included in this review of microbeam systems for cell irradiation at RARAF.

Multiphoton ionization of many-electron atoms and highly-charged ions in intense laser fields: a relativistic time-dependent density functional theory approach

Science.gov (United States)

Tumakov, Dmitry A.; Telnov, Dmitry A.; Maltsev, Ilia A.; Plunien, Günter; Shabaev, Vladimir M.

2017-10-01

We develop an efficient numerical implementation of the relativistic time-dependent density functional theory (RTDDFT) to study multielectron highly-charged ions subject to intense linearly-polarized laser fields. The interaction with the electromagnetic field is described within the electric dipole approximation. The resulting time-dependent relativistic Kohn-Sham (RKS) equations possess an axial symmetry and are solved accurately and efficiently with the help of the time-dependent generalized pseudospectral method. As a case study, we calculate multiphoton ionization probabilities of the neutral argon atom and argon-like xenon ion. Relativistic effects are assessed by comparison of our present results with existing non-relativistic data.

Ultra slow muon microscopy by laser resonant ionization at J-PARC, MUSE

Energy Technology Data Exchange (ETDEWEB)

Miyake, Y., E-mail: yasuhiro.miyake@kek.jp; Ikedo, Y.; Shimomura, K.; Strasser, P.; Kawamura, N.; Nishiyama, K.; Koda, A.; Fujimori, H.; Makimura, S.; Nakamura, J.; Nagatomo, T.; Kadono, R. [High Energy Accelerator Research Organization (KEK), Muon Science Laboratory (Japan); Torikai, E. [Yamanashi University, Faculty of Engineering (Japan); Iwasaki, M. [RIKEN Nishina Center, Advanced Meson Science Laboratory (Japan); Wada, S.; Saito, N. [RIKEN, Advanced Science Institute (Japan); Okamura, K. [RIKEN-WAKO Incubation Plaza 301, Megaopto Co., Ltd. (Japan); Yokoyama, K. [RIKEN Nishina Center, Advanced Meson Science Laboratory (Japan); Ito, T.; Higemoto, W. [J-PARC Center, Muon Section, Materials and Life Science Division (Japan)

2013-04-15

As one of the principal muon beam line at the J-PARC muon facility (MUSE), we are now constructing a Muon beam line (U-Line), which consists of a large acceptance solenoid made of mineral insulation cables (MIC), a superconducting curved transport solenoid and superconducting axial focusing magnets. There, we can extract 2 Multiplication-Sign 10{sup 8}/s surface muons towards a hot tungsten target. At the U-Line, we are now establishing a new type of muon microscopy; a new technique with use of the intense ultra-slow muon source generated by resonant ionization of thermal Muonium (designated as Mu; consisting of a {mu}{sup + } and an e{sup - }) atoms generated from the surface of the tungsten target. In this contribution, the latest status of the Ultra Slow Muon Microscopy project, fully funded, is reported.

Gas chromatography/multiphoton ionization/time-of-flight mass spectrometry of polychlorinated biphenyls

International Nuclear Information System (INIS)

Matsui, Taiki; Uchimura, Tomohiro; Imasaka, Totaro

2011-01-01

A sample mixture of polychlorinated biphenyls (PCBs) was measured by gas chromatography/multiphoton ionization/time-of-flight mass spectrometry (GC/MPI/TOF-MS) using four types of laser sources. When a fourth harmonic emission (266 nm) of a picosecond Nd:YAG laser (1064 nm) was utilized, highly chlorinated PCBs larger than hepta-CBs were not observed. A fifth harmonic emission (213 nm) of the picosecond Nd:YAG laser allowed the measurement of PCBs from di-CBs to octa-CBs, and the limit of detection (LOD) was several pg for each component of PCBs. The LOD for the total amount of PCBs, which was calculated using the protocol provided by the Ministry of the Environment, Japan, was 1000 pg. The signal intensity of the congeners with chlorine atoms at the ortho positions (non-coplanar PCBs) was enhanced by using the fifth harmonic emission. When the fourth harmonic emission remaining after fifth harmonic generation was simultaneously used, the LOD for total PCBs was improved to 667 pg. The PCB sample was also measured using a third harmonic emission (267 nm) of a femtosecond Ti:sapphire laser (800 nm), providing an LOD of 677 pg. Thus, the two-color beam (266/213 nm) of a picosecond Nd:YAG laser had a comparable, or even slightly superior, performance to the more expensive femtosecond Ti:sapphire laser.

Laser stimulation can activate autophagy in HeLa cells

International Nuclear Information System (INIS)

Wang, Yisen; Hu, Minglie; Wang, Chingyue; Lan, Bei; Cao, Youjia; He, Hao

2014-01-01

For decades, lasers have been a daily tool in most biological research for fluorescent excitation by confocal or multiphoton microscopy. More than 20 years ago, cell photodamage caused by intense laser stimulation was noticed by generating reactive oxygen species, which was then thought as the main damage effect by photons. In this study, we show that laser stimulation can induce autophagy, an important cell lysosomal pathway responding to immune stimulation and starvation, without any biochemical treatment. Two different types of laser stimulations are found to be capable of activating autophagy: continuous scanning by continuous-wave visible lasers and a short-time flash of femtosecond laser irradiation. The autophagy generation is independent from wavelength, power, and scanning duration of the visible lasers. In contrast, the power of femtosecond laser is very critical to autophagy because the multiphoton excited Ca 2+ dominates autophagy signaling. In general, we show here the different mechanisms of autophagy generation by such laser stimulation, which correspond to confocal microscopy and cell surgery, respectively. Those results can help further understanding of photodamage and autophagy signaling.

A microwave-assisted solution combustion synthesis to produce europium-doped calcium phosphate nanowhiskers for bioimaging applications.

Science.gov (United States)

Wagner, Darcy E; Eisenmann, Kathryn M; Nestor-Kalinoski, Andrea L; Bhaduri, Sarit B

2013-09-01

Biocompatible nanoparticles possessing fluorescent properties offer attractive possibilities for multifunctional bioimaging and/or drug and gene delivery applications. Many of the limitations with current imaging systems center on the properties of the optical probes in relation to equipment technical capabilities. Here we introduce a novel high aspect ratio and highly crystalline europium-doped calcium phosphate nanowhisker produced using a simple microwave-assisted solution combustion synthesis method for use as a multifunctional bioimaging probe. X-ray diffraction confirmed the material phase as europium-doped hydroxyapatite. Fluorescence emission and excitation spectra and their corresponding peaks were identified using spectrofluorimetry and validated with fluorescence, confocal and multiphoton microscopy. The nanowhiskers were found to exhibit red and far red wavelength fluorescence under ultraviolet excitation with an optimal peak emission of 696 nm achieved with a 350 nm excitation. Relatively narrow emission bands were observed, which may permit their use in multicolor imaging applications. Confocal and multiphoton microscopy confirmed that the nanoparticles provide sufficient intensity to be utilized in imaging applications. Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Laser stimulation can activate autophagy in HeLa cells

Energy Technology Data Exchange (ETDEWEB)

Wang, Yisen; Hu, Minglie; Wang, Chingyue [Ultrafast Laser Laboratory, Key Laboratory of Optoelectronic Information Technology (Ministry of Education), College of Precision Instrument and Optoelectronics Engineering, Tianjin University, Tianjin (China); Lan, Bei; Cao, Youjia [Key Laboratory of Microbial Functional Genomics of Ministry of Education, College of Life Sciences, Nankai University, Tianjin (China); He, Hao, E-mail: haohe@tju.edu.cn [Ultrafast Laser Laboratory, Key Laboratory of Optoelectronic Information Technology (Ministry of Education), College of Precision Instrument and Optoelectronics Engineering, Tianjin University, Tianjin (China); Med-X Research Institute, School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai (China)

2014-10-27

For decades, lasers have been a daily tool in most biological research for fluorescent excitation by confocal or multiphoton microscopy. More than 20 years ago, cell photodamage caused by intense laser stimulation was noticed by generating reactive oxygen species, which was then thought as the main damage effect by photons. In this study, we show that laser stimulation can induce autophagy, an important cell lysosomal pathway responding to immune stimulation and starvation, without any biochemical treatment. Two different types of laser stimulations are found to be capable of activating autophagy: continuous scanning by continuous-wave visible lasers and a short-time flash of femtosecond laser irradiation. The autophagy generation is independent from wavelength, power, and scanning duration of the visible lasers. In contrast, the power of femtosecond laser is very critical to autophagy because the multiphoton excited Ca{sup 2+} dominates autophagy signaling. In general, we show here the different mechanisms of autophagy generation by such laser stimulation, which correspond to confocal microscopy and cell surgery, respectively. Those results can help further understanding of photodamage and autophagy signaling.

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

Science.gov (United States)

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

2013-02-01

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

Dynamics of NO2 dissociation. Study by resonance-enhanced multi-photon ionisation of energy distribution and of anisotropies in fragments

International Nuclear Information System (INIS)

Mons, Michel

1988-01-01

In this research thesis, the author reports the use of laser resonance-enhanced multi-photon ionization and of time-of-flight mass spectrometry for a detailed characterization of fragments produced by a photo-dissociation process. The author more particularly addressed the case of a NO 2 molecule excited at low energies above the dissociation threshold. In the first part, the author discusses issues and problems related to molecular photo-dissociation. In the second part, he presents the developed method and shows that the combined use of both techniques allows a precise characterisation of photo-fragments in terms of internal or translational energies as well as in terms of angle distributions. Finally, the author presents and discusses results obtained in the case of NO 2 [fr

Silicon isotope separation utilizing infrared multiphoton dissociation of Si2F6 irradiated with two-color CO2 laser light

International Nuclear Information System (INIS)

Yokoyama, Atsushi; Ohba, Hironori; Hashimoto, Masashi; Arai, Shigeyoshi

2002-01-01

Silicon isotope separation has been done by utilizing the Infrared Multiphoton Dissociation (IRMPD) of Si 2 F 6 irradiated with two-color CO 2 laser lights. The two-color excitation method improved the separation efficiency keeping the high enrichment factors. For example, 99.74% of 28 Si was obtained at 49.63% dissociation of Si 2 F 6 after the simultaneous irradiation of 200 pulses with 966.23 cm -1 photons (0.084 J/cm 2 ) and 954.55 cm -1 photons (0.658 J/cm 2 ), while 2000 pulses were needed to obtain 99.35% of 28 Si at 35.6% dissociation in the case of only one-color irradiation at 954.55 cm -1 (0.97 J/cm 2 ). (author)