... Title : Multiphoton and Multicolor Confocal Microscopy of Microstructure, Fracture Delamination and Thermal Degradation in Multilayer Organic ...

Intravital multiphoton microscopy provides a unique opportunity to discover and characterize biological phenomena in the natural context of living organisms. Here we provide an overview of multiphoton microscopy with particular attention to its application for studying host-pathogen interactions.

The next evolution in multiphoton microscopy will further enhance the ability to observe complex and dynamic biological processes from deeper within living tissue with minimal invasion and photodamage.

... Accession Number : ADA077227. Title : Mechanisms of Multiphoton Dissociation of Molecular Ions. Descriptive Note : Summary rept. ...

... Accession Number : ADA120570. Title : Detection of Rocket Fuel Components by Multiphoton Photoionization Mass Spectroscopy. ...

... Accession Number : ADA136446. Title : Competing Dissociation Channels in the Infrared Multiphoton Decomposition of Ethyl Vinyl Ether,. ...

... Accession Number : ADA116057. Title : Comparison of Multiphoton Ionization Fragmentation Patterns for the Azulene and Naphthalene Isomers. ...

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 ...

... conference papers discuss work in the areas of biomedical imaging, microscopy, spectroscopy, fluorescence, multiphoton microscopy, fiber optics ...

Intravital multiphoton microscopy has provided powerful mechanistic insights into health and disease, and has become a common instrument in the modern biological laboratory. The requisite high numerical aperture and exogenous contrast agents that enable multiphoton microscopy, however, limit ability to investigate ...

We present a combined multiphoton-acoustic microscope giving collocated access to the local morphological as well as mechanical properties of living cells. Both methods relay on intrinsic contrast mechanisms and dispense with the need of staining. In the acoustic part of the microscope, a gigahertz ultrasound wave is generated by an acoustic lens and the ...

Typical imaging depths with multiphoton microscopy (MPM) are limited to less than 300 mum in many tissues due to light scattering. Optical clearing significantly reduces light scattering by replacing water in the organ tissue with a fluid having a similar index of refraction to that of proteins. We demonstrate MPM of intact, fixed, cleared mouse organs ...

order), except BTI people. Oracle Calendar for NBTC Confocal / Multiphoton Microscope httpNBTC Confocal / Multiphoton Microscopy Facility D21-B and D22 Clark Hall (revised 8/06) Carol Confocal and Multiphoton System is located in the CCMR Materials Facility D21 Clark Hall and the office

The accessibility of tunable, ultrafast laser sources has spurred the development and wide application of specialized microscopy techniques based on chromophore fluorescence following two- and three-photon absorption. The attendant advantages of such methods, which have led to a host of important applications including three-dimensional biological imaging, ...

The dynamic ability of neuronal dendrites to shape and integrate synaptic responses is the hallmark of information processing in the brain. Effectively studying this phenomenon requires concurrent measurements at multiple sites on live neurons. Significant progress has been made by optical imaging systems which combine confocal and multiphoton microscopy ...

... Accession Number : AD0760800. Title : Note Concerning Multi-Photon Ionization by a Laser Beam,. Corporate Author : INSTITUTE ...

... Accession Number : ADP011222. Title : Multi-Photon Fluorescence Spectroscopy of Fluorescent Bio-Probes and Bio-Molecules. ...

... Accession Number : ADA385825. Title : Multi-Photon Entanglement and Quantum Teleportation. Descriptive Note : Final rept. ...

... Accession Number : ADA099121. Title : Mechanisms of Multiphoton Dissociation of Molecular Ions. Descriptive Note : Final rept. Sep 76-Mar 81,. ...

Multiphoton microscopy is a powerful technique based on complex quantum mechanical effects. Thanks to the development of turnkey mode-locked laser systems, multiphoton microscopy is now available for everyone to use without extreme complexity. In this short introduction, we describe qualitatively the important ...

of the broadband laser pulse is extracted analytically with double quadrature spectral interferometry (DQSICharacterization of a broadband pulse for phase controlled multiphoton microscopy by single beam beam. Two narrow pulses and one broadband pulse are selected out of an ultrafast laser pulse

The research in biomedical photonics is clearly evolving in the direction of the understanding of biological processes at the cell level. The spatial resolution to accomplish this task practically requires photonics tools. However, an integration of different photonic tools and a multimodal and functional approach will be necessary to access the mechanical and biochemical cell ...

Multiphoton excitation of molecular probes has become an important tool in experimental neurobiology owing to the intrinsic optical sectioning and low light scattering it affords. Using molecular functional indicators, multiphoton excitation allows physiological signals within single neurons to be observed from within living brain tissue. Ideally, it would ...

We review the current state of multiphoton microscopy. In particular, the requirements and limitations associated with high-speed multiphoton imaging are considered. A description of the different scanning technologies such as line scan, multifoci approaches, multidepth microscopy, and novel detection techniques is ...

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 ...

Multiphoton microscopy has enabled biologists to collect high-resolution images hundreds of microns into biological tissues, including tissues of living animals. While the depth of imaging exceeds that possible from any other form of light microscopy, multiphoton microscopy is nonetheless ...

The spectroscopic content within optical coherence tomography (OCT) data can provide a wealth of information. Spectroscopic OCT methods are frequently limited by time-frequency trade-offs that limit high spectral and spatial resolution simultaneously. We present spectroscopic spectral-domain optical coherence microscopy performed with a multimodality microscope. Restricting ...

Unlike conventional multiphoton excited microscopy according to pixel-by-pixel point scanning, a widefield multiphoton excited microscopy based on spatiotemporal focusing has been developed to construct three-dimensional (3D) multiphoton fluorescence images only with the need of an axial ...

Currently we designed and implemented a combined spectral and lifetime imaging system which combines the benefits of multiphoton microscopy, spectral discrimination, lifetime analysis and temporal information. This system has efficiencies of > 50% and cap...

The application of multiphoton microscopy to the biological sciences has led to a new generation of imaging-based studies extending from the tracking of individual molecules within living cells to the observation of whole organisms

The use of multiphoton microscopy for imaging mouse brain in vivo offers several advantages and poses several challenges. This tutorial begins by briefly comparing multiphoton microscopy with other imaging modalities used to visualize the brain and its activity. Next, an overview of the techniques for introducing ...

Gradient index lenses enable multiphoton microscopy of deep tissues in the intact animal. In order to assess their applicability to clinical research, we present in vivo multiphoton microscopy with gradient index lenses in brain regions associated with Alzheimer's disease and Parkinson's disease in both transgenic ...

Reflected confocal microscopy has been widely used in clinical application in dermatology. In recent years, multiphoton microscopy has also emerged as an important minimally invasive bioimaging technique for the skin. In this study, we combine reflected confocal microscopy and multiphoton ...

Multi-photon microscopy, now in its twentieth year, has developed into one of the most robust and powerful techniques for live cell and in vivo fluorescence imaging. Although its theoretical framework is nearly a century old, it has only become a practical tool for biological research with the development of ultrafast lasers and scanning ...

Background & AimsThe ability to observe cellular and subcellular detail during routine endoscopy is a major goal in the development of new endoscopic imaging techniques. Multiphoton microscopy, which relies on nonlinear infared optical processes, has the potential to identify cellular details by excitation of endogenous fluorescent molecules. We ...

In a typical multiphoton microscope, the majority of the laser power from the near-IR pulsed source is thrown away in order to prevent photo-damage on biological samples. It is thus desirable to convert this wasted power into other wavelengths through nonlinear fiber optics to accommodate imaging modalities beyond conventional multiphoton ...

We have developed a new descanned parallel (32-fold) pinhole and photomultiplier detection array for multifocal multiphoton microscopy that effectively reduces the blurring effect originating from scattered fluorescence photons in strongly scattering biological media. With this method, we achieve a fourfold improvement in photon statistics for detecting ...

We propose and demonstrate a dynamic point spread function (PSF) for single and multiphoton fluorescence microscopy. The goal is to generate a PSF whose shape and size can be maneuvered from highly localized to elongated one, thereby allowing shallow-to-depth excitation capability during active imaging. The PSF is obtained by utilizing specially designed ...

... Accession Number : ADP007081. Title : Multiphoton Ionization of I2, Hi, and Cf3i, and Above-Threshold Ionization inI2,. Corporate ...

... Accession Number : ADA183703. Title : Infrared Multiphoton Dissociation of RDX (Hexahydro-1,3,5-trinitro-1,3,5-triazine) in a Molecular Beam. ...

studies of some addictive drugs following dansyl chloride labelling, Talanta 43 (1996), 1429�1436. 8 #12;

... We have used maize leaf protoplast as a model system to evaluate the photo-induced response of living sample under high intensity illumination. ...

Collinear pulse measurement tools useful at the full numerical aperture (NA) of multiphoton microscope objectives are a necessity for a quantitative characterization of the femtosecond pulses focused by these systems. In this letter, we demonstrate a simple new technique, for characterizing the pulse at the focus in a multiphoton microscope. This ...

In multiphoton fluorescence laser-scanning microscopy, ultrafast laser pulses [i.e., light pulses having pulse width multiphoton absorption cross-sections of common fluorophores. Because of the broad overlapping two-photon absorption spectra of fluorophores and ...

In multiphoton fluorescence laser-scanning microscopy, ultrafast laser pulses [i.e., light pulses having pulse width <=1 ps (1 ps=10-12 s)] are commonly employed to circumvent the low-multiphoton absorption cross-sections of common fluorophores. Because of the broad overlapping two-photon absorption spectra of fluorophores and the ...

In this work, intravital multiphoton microscopy is applied to the imaging of liver metabolism with the least invasiveness. We observed intravital dynamics of the uptake, processing and excretion of the organic anion species, 6-carboxyfluorescein diacetate (6-CFDA) in the hepatobiliary system. This is achieved by the use of multiphoton ...

We present the application of remote focusing to multiphoton laser scanning microscopy and utilize this technology to demonstrate simultaneous, programmable multi-layer imaging. Remote focusing is used to independently control the axial location of multiple focal planes that can be simultaneously imaged with single element detection. This ...

Conventional diagnosis for hepatocellular carcinoma (HCC) is time consuming in sample preparation and deficient in quantitative examination. In this work, we developed multiphoton autofluorescence (MAF) microscopy as an effective and efficient tool in HCC diagnosis with qualitative imaging and quantitative measurement. We found that MAF imaging effectively ...

... Accession Number : ADA262301. Title : Scanning Tunneling Microscopy, Atomic Force Microscopy, and Related Techniques. ...

Multiphoton laser scanning microscopy (MPLSM) is a promising tool to study the tissue distribution of environmental chemical contaminants during fish early life stages. One such chemical for which this is possible is benzo[a]pyrene (BaP), a polycyclic aromatic hydrocarbon that a...

While multiphoton microscopy (MPM) has been performed with a wide range of excitation wavelengths, fluorescence emission has been limited to the visible spectrum. We introduce a paradigm for MPM of near-infrared (NIR) fluorescent molecular probes via nonlinear excitation at 1550 nm. This all-NIR system expands the range of available MPM fluorophores, ...

Multiphoton laser scanning microscopy (MPLSM) has proved to be an important tool in cancer research providing insight into morphologic changes in tumor vasculature, extracellular matrix components and gene expression. However, there is a lack of technique...

Intravital microscopy (IVM) allows for the direct in vivo visualization of dynamic biological processes in their physiological context at high spatial and temporal resolution. Novel nonlinear optical imaging modalities, most prominently multiphoton microscopy, have extended the spectrum of cellular functions amenable to IVM ...

... SW Chu, TM Liu, PC Cheng,"High intensity scanning microscopy with a femtosecond Cr:Forsterite laser," Scanning, 22, pp. 95-96,2000. 4. K. Konig ...

We use a novel nonlinear optical technique, multiphoton microscopy, to differentiate keloids from hypertrophic and normal scars through the morphological and quantitative analysis of collagen and elastin in superficial dermis of scar tissues and boundary between normal skin and scar tissue.

Multiphoton microscopy has been added to the array of imaging techniques at the endstation for the Microbeam II cell irradiator at Columbia University�s Radiological Research Accelerator Facility (RARAF). This three-dimensional (3D), laser-scanning microscope functions through multiphoton excitation, providing an enhanced imaging ...

By means of multiphoton laser scanning microscopy, neuroscientists can look inside the brain deeper than has ever been possible before. Multiphoton fluorescent images, as all optical images, suffer from degradation caused by a variety of sources (e.g. light dispersion and absorption in the tissue, laser fluctuations, spurious ...

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, ...

The aim of this work is to demonstrate that multiphoton microscopy is a preferred technique to investigate intact cornea structure without slicing and staining. At the micron resolution, multiphoton imaging can provide both large morphological features and detailed structure of epithelium, corneal collagen fibril bundles and ...

The use of semiconductor nanocrystals (quantum dots) as fluorescent labels for multiphoton microscopy enables multicolor imaging in demanding biological environments such as living tissue. We characterized water-soluble cadmium selenide-zinc sulfide quantum dots for multiphoton imaging in live animals. These fluorescent probes have ...

The aim of this study is to assess the application of multiphoton autofluorescence and second harmonic generation (SHG) microscopy for investigating corneal wound healing after high myopic (-10.0D) photorefractive keratectomy (PRK) procedures on the rabbit eyes. The effect of PRK on the morphology and distribution of keratocytes were investigated using ...

Lamina propria (LP) of gastric mucosa plays an important role in progression of gastric cancer because of the site at where inflammatory reactions occur. Multiphoton imaging has been recently employed for microscopic examination of intact tissue. In this paper, using multiphoton microscopy (MPM) based on two-photon excited fluorescence ...

Intravital imaging emerged as an indispensible tool in biological research, and a variety of imaging techniques have been developed to noninvasively monitor tissues in vivo. However, most of the current techniques lack the resolution to study events at the single-cell level. Although intravital multiphoton microscopy has addressed this limitation, the need ...

A microfluidic mixer is applied to study the kinetics of calmodulin conformational changes upon Ca²⁺ binding. The device facilitates rapid, uniform mixing by decoupling hydrodynamic focusing from diffusive mixing and accesses time scales of tens of microseconds. The mixer is used in conjunction with multiphoton ...

For the last decade, multiphoton excitation fluorescence microscopy has found numerous applications in biology. Multiphoton microscopy provides several advantages over conventional fluorescence microscopy, including increased penetration depth, improved signal-to-background ratio, and reduced ...

We recorded one-photon excited fluorescence (1PEF) and two-photon excited fluorescence (2PEF) spectra of purified keratin from human epidermis, and determined the action cross section of this endogenous chromophore. We used this spectroscopic analysis to analyse multiphoton images of skin biopsies and assign the intrinsic fluorescence signals in the epidermis. We observed a ...

Photobleaching as a result of intense laser irradiation has been an important issue in the applications of confocal microscopy. In this study we are employing time-lapsed micro-spectroscopy to compare the effects of photobleaching under single photon and multi-photon excitation. A model is derived to account for photobleaching in both cases and protoplasts ...

We develop an adaptive optics (AO) multiphoton microscope by incorporating a deformable mirror and a Hartmann-Shack wavefront sensor. The AO module operating in closed-loop is used to correct for the aberrations of the illumination laser beam. This increases the efficiency of the nonlinear processes in reducing tissue photodamage, improves contrast, and enhances lateral ...

We develop an adaptive optics (AO) multiphoton microscope by incorporating a deformable mirror and a Hartmann-Shack wavefront sensor. The AO module operating in closed-loop is used to correct for the aberrations of the illumination laser beam. This increases the efficiency of the nonlinear processes in reducing tissue photodamage, improves contrast, and enhances lateral ...

An integrated microscope that combines different optical techniques for simultaneous imaging is demonstrated. The microscope enables spectral-domain optical coherence microscopy based on optical backscatter, and multiphoton microscopy for the detection of two-photon fluorescence and second harmonic generation signals. The unique ...

We present combined coherent anti Stokes Raman scattering and two-photon excited fluorescence microscopy, for label-free in vitro and in vivo imaging of skin with both chemical discrimination and subcellular resolution. In particular, we show how CARS microscopy is utilized to image lipid-rich structures inside the skin, preparing for a combined ...

The need for quantification and real time visualization of developmental processes has called for increasingly sophisticated imaging techniques. Among them, multiphoton microscopy reveals itself to be an extremely versatile tool owing to its unique ability to combine fluorescent imaging, laser ablation, and higher harmonic generation. Furthermore, recent ...

By introducing spatiotemporal pulse shaping techniques to multiphoton microscopy it is possible to obtain video-rate images with depth resolution similar to point-by-point scanning multiphoton microscopy while mechanically scanning in only one dimension. This is achieved by temporal focusing of the illumination ...

We performed intravital multiphoton microscopy to image and analyze normal and carcinogen treated skin tissues of nude mice in vivo. Using intravital images and the quantitative pixel to pixel ratiometric processing of multiphoton autofluorescence to second harmonic generation index (MAFSI), we can visualize the interaction between ...

Multiphoton imaging is a promising approach for addressing current issues in systems biology and high-content investigation of embryonic development. Recent advances in multiphoton microscopy, including light-sheet illumination, optimized laser scanning, adaptive and label-free strategies, open new opportunities for embryo imaging. ...

Population dynamics of patterned neuronal firing are fundamental to information processing in the brain. Multiphoton microscopy in combination with calcium indicator dyes allows circuit dynamics to be imaged with single-neuron resolution. However, the temporal resolution of fluorescent measures is constrained by the imaging frequency imposed by standard ...

The purpose of this study is to assess the application of multiphoton fluorescence and second harmonic generation (SHG) microscopy for imaging and monitoring the disease progress of infectious keratitis in an experimental model, and to investigate the possible correlation of tissue architecture with spreading patterns of pathogens in an experimental model. ...

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 ...

Multiphoton laser scanning micrsocopy holds promise as a tool to study the tissue distribution of environmental chemical contaminants during fish early life stage development. One such chemical for which this is possible is benzo[a]pyrene (BaP), a polyaromatic hydrocarbon that a...

or in vivo conditions, two-photon microscopy has also been recently applied to investigating transdermal trans- port pathways of hydrophilic and hydrophobic molecules under chemical enhancer actions (Yu et al into improving drug delivery methodology. In-depth, multi-photon examination of bio- logical specimen is now

By using two-photon autofluorescence (TPAF) and second harmonics generation (SHG), we imaged hepatocellular carcinoma (HCC) biopsies from the human patients and compared them with the conventional histological biopsies. We found that multiphoton microscopy may be used to obtain label-free images of liver tissues and may be developed into an effective ...

Journal of Biomedical Optics 15(6), 066004 (November/December 2010) Adaptive optics multiphoton Laboratorio de �Optica Campus de Espinardo (CiOyN) Murcia, 30100 Spain Abstract. We develop an adaptive optics; adaptive optics; human ocular tissue. Paper 10192PRR received Jun. 8, 2010; revised manuscript received Aug

Combining multiphoton microscopy with a newly designed hepatic imaging window, we acquired in vivo images of mice obstructive cholestasis. We observed that in mice with bile duct ligation, bile canaliculi failed to appear during the whole observation period over 100 minutes following carboxyfluorescein diacetate injection, whereas the fluorescence was ...

... Accession Number : AD0715516. Title : Dry Mass Determination by Interference Microscopy; Agreement with Quantitative Electron Microscopy,. ...

... Accession Number : AD0194211. Title : ULTRASONIC ABSORPTION AND MICROSCOPY AND SPECTROSCOPY. Corporate Author : ...

... Accession Number : AD0708629. Title : TRANSMISSION ELECTRON MICROSCOPY OF SODIUM CHLORIDE,. Corporate ...

... Accession Number : AD0834894. Title : SCANNING ELECTRON MICROSCOPY STUDIES OF DENTAL ENAMEL. Descriptive ...

... Accession Number : AD0664993. Title : FEASIBILITY OF MICROSCOPY FOR INVESTIGATING THE SILVER OXIDE ELECTRODE. ...

We demonstrate how optical tweezers can be incorporated into a multiphoton microscope to achieve three-dimensional imaging of trapped cells. The optical tweezers, formed by a cw 1064 nm Nd:YVO4 laser, were used to trap live yeast cells in suspension while the 4',6-diamidino-2-phenylindole-stained nucleus was imaged in three dimensions by use of a pulsed femtosecond laser. The ...

Recently, multiphoton microscopy has gained much popularity as a noninvasive imaging modality in biomedical research. We evaluate the potential of multiphoton microscopy for monitoring laser-skin reaction in vivo. Nude mouse skin is irradiated with an erbium:YAG laser at various fluences and immediately imaged by a ...

Recently, multiphoton microscopy has gained much popularity as a noninvasive imaging modality in biomedical research. We evaluate the potential of multiphoton microscopy for monitoring laser-skin reaction in vivo. Nude mouse skin is irradiated with an erbium:YAG laser at various fluences and immediately imaged by a ...

Real-time in vivo imaging of molecular targets at (sub)cellular resolution is essential in better understanding complex biology. Confocal microscopy and multiphoton microscopy have been used in the past to achieve this goal, but their true capabilities have often been limited by bulky optics and difficult experimental set-ups requiring ...

AbstractReal-time in vivo imaging of molecular targets at (sub)cellular resolution is essential in better understanding complex biology. Confocal microscopy and multiphoton microscopy have been used in the past to achieve this goal, but their true capabilities have often been limited by bulky optics and difficult experimental set-ups ...

In these studies, the potential of optical techniques for early breast cancer detection were tested in animal models and cell culture. Optical spectroscopy, in vivo and in vitro microscopy studies indicate that optical methods show great promise for the e...

Multiphoton microscopy is a techniques that fascilitates three dimensional imaging of intact, unstained tissue. Especially connective tissue has a relatively strong nonlinear optical response and can easily be imaged. Atherosclerosis is a disease where lipids accumulate in the vessel wall and there is a thickening of the intima by growth of a cap of ...

Multiphoton laser scanning microscopy (MPLSM) has become an increasingly invaluable tool in fluorescent optical imaging. There are several distinct advantages to implementing MPLSM as a Xenopus oocyte research tool. MPLSM increases signal-to-noise ratio and therefore increases image quality because there is no out-of-focus fluorescence as would be created ...

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To assess laser therapies in clinical practice, histologic examinations were commonly used. But histologic examinations were invasive and not real-time in nature. In this work, we validate multiphoton microscopy as a useful modality in evaluating laser-tissue reaction in vivo. Multiphoton microscopy based on ...

... and pH in Living Breast Tumor Models with Three-Dimensional Resolution by Multiphoton Microscopy during Combined Therapy with Herceptin. ...

The aim of this study is to characterize the ability of multiphoton microscopy in monitoring the transdermal penetration enhancing effect of a depilatory agent and the associated structural alterations of stratum corneum. The result is compared with the electron microscopic findings. Our results show that the penetration of both hydrophilic and hydrophobic ...

The potential of multiphoton microscopy in providing in-vivo early diagnosis of skin lesions has already been demonstrated, while its capability in therapy follow-up has not been deeply explored so far. Two-photon excited fluorescence and second-harmonic generation microscopy were used in combination to follow-up collagen remodeling ...

Multiphoton microscopy was employed for monitoring the structure changes of mouse dermis collagen in the intrinsic- or the extrinsic-age-related processes in vivo. The characteristics of textures in different aging skins were uncovered by fast Fourier transform in which the orientation index and bundle packing of collagen were quantitatively analyzed. Some ...

A factor that limits the use of multiphoton microscopy (MPM) in clinical and preclinical studies is the lack of a compact and flexible probe. We report on a miniaturized MPM probe employing a microelectromechanical system (MEMS) scanning mirror and a double-clad photonic crystal fiber (DCPCF). The use of a MEMS mirror and a DCPCF provides many advantages, ...

Multiphoton microscopy (MPM) has high resolution and sensitivity. In this study, MPM was used to image microstructure of human rectal mucosa. The morphology and distribution of the main components in mucosa layer, absorptive cells and goblet cells in the epithelium, abundant intestinal glands in the lamina propria and smooth muscle fibers in the muscularis ...

We demonstrate high-resolution fluorescence microscopy based on a cyclic sequential multiphoton (CSM) process, which gives rise to fluorescence emission following a sequence of cyclic transitions between the bright and dark states of a fluorophore induced by pump and reverse light. By temporally modulating the reverse intensity, we can extract the ...

Using multiphoton autofluorescence (MAF) and second harmonic generation (SHG) microscopy, we investigate the morphology and the structure of the corneal epithelium and stroma collagen of bovine cornea following injection of Pseudomonas aeruginosa. We found that corneal epithelial cells are damaged and stromal collagen becoming increasingly autofluorescent ...

Three-dimensional (3D) resolution improvement in multi-photon multiple-excitation-spot-optical microscopy is proposed. Specially designed spatial filter is employed for improving the overall 3D resolution of the imaging system. An improvement up to a factor of 14.5 and sub-femto liter volume excitation is achieved. The system shows substantial sidelobe ...

Three-dimensional (3D) resolution improvement in multi-photon multiple-excitation-spot-optical microscopy is proposed. Specially designed spatial filter is employed for improving the overall 3D resolution of the imaging system. An improvement up to a factor of 14.5 and sub-femto liter volume excitation is achieved. The system shows substantial sidelobe ...

Standard multiphoton laser scanning microscopy (MPLSM) has revolutionized our view of physiologic and pathologic processes in living organisms by enlightening different aspects of cellular choreography in immune responses, that is, cellular motility and co-localization. To understand cellular communication on a molecular level, novel transgenic reporter ...

Tissue glycation from diabetes and aging can result in complications such as renal failure, blindness, nerve damage and vascular diseases. In this work, we applied multiphoton microscopy for imaging and characterizing the extent of tissue glycation. The characteristic features of multiphoton autofluorescence (MPAF) and second harmonic ...

We have investigated lipid acyl chain order parameters of giant unilamellar vesicles (GUVs) using multiphoton fluorescence microscopy. We compare two widely used models of lipid acyl chain order parameters: the "wobble-on-a-cone" model and the Gaussian distribution model. For the first time, we systematically address a ternary system for which the phase ...

Abstract: Real-time histology or virtual biopsy for the diagnosis of colonic cancer is of great medical significance. In this work, we show that label-free multiphoton imaging is feasible and effective in monitoring colonic cancer progression by providing cellular and subcellular details in fresh, unfixed, unstained colonic specimens. Our results also demonstrate the ...

We performed multiphoton excited autofluorescence and second harmonic generation microscopy for the distinction of normal, lung adenocarcinoma (LAC), and squamous cell carcinoma (SCC) specimens. In addition to morphological distinction, we derived quantitative metrics of cellular redox ratios for cancer discrimination. Specifically, the redox ratios of ...

multiphoton microscopy, Opt. Lett. 23:655�657. Bewersdorf, J., Schmidt, R., and Hell, S.W., 2005, Comparison microscopy and I5 M, coherently combines the aperture of two opposing lenses (Hell and Stelzer, 1992a emission depletion (STED) microscopy (Hell and Wichmann, 1994; see also Chapter 31, this volume

The design and operation of a time-of-flight mass analyzer for study of ionization and dissociation of atoms and molecules produced by interaction with pulsed laser generated by an OPO are described. This apparatus allows the investigation of atomic and molecular excited states no accessible by single-photon absorption. The description of the main parts as well as the ...

Detailed morphological and cellular information relating to the human tooth have traditionally been obtained through histological studies that required decalcification, staining, and fixation. With the recent invention of multiphoton microscopy, it has become possible to acquire high resolution images without histological procedures. Using an ...

Multiphoton laser scanning microscopy offers advantages in depth of penetration into intact samples over other optical sectioning techniques. To achieve these advantages it is necessary to detect the emitted light without spatial filtering. In this nondescanned (nonconfocal) approach, ambient room light can easily contaminate the signal, forcing ...

Translating discoveries made in isolated renal cells and tubules to the in vivo situation requires the assessment of cellular function in intact live organs. Multiphoton imaging is a form of fluorescence microscopy that is ideally suited to working with whole tissues and organs, but adequately loading cells with fluorescence dyes in vivo remains a ...

A powerful advantage of multiphoton microscopy is its ability to image endogenous fluorophores such as the ubiquitous coenzyme NADH in discrete cellular populations. NADH is integral in both oxidative and non-oxidative cellular metabolism. NADH loses fluorescence upon oxidation to NAD+; thus changes in NADH fluorescence can be used to monitor metabolism. ...

We describe multimodal nonlinear microscopy using a compact, turnkey femtosecond fiber laser at 1.5 ?m. The system allows for multiplexed detection of near infrared and visible contrast agents through two-and three-photon excitation fluorescence microscopy as well as structural imaging viaauto-confocal microscopy (ACM). This platform ...

For the last two decades, multiphoton excitation microscopy/microfabrication based on laser scanning/writing techniques has been popular in the life science as well as photonics. Due to the slow scanning/writing nature, these applications are very limited to the production of prototypes, although its submicron optical resolution and intrinsic 3D optical ...

A handful of research teams around the world have recently begun to utilize multiphoton techniques in cytometry, especially for in vivo applications. These approaches offer similar enhancements to flow cytometry as the multiphoton phenomenon brought to the field of microscopy at the turn of the 20th century, with at least six ...

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 ...

Fluorescence lifetime imaging microscopy (FLIM) using multiphoton excitation is emerging as a reliable quantitative tool for measuring fluorescence resonance energy transfer (FRET) in living cells. By virtue of being free from spectroscopic artifacts encountered in conventional FRET detection methods, multiphoton FLIM methods offer the ...

Conductive keratoplasty (CK) is a new refractive surgery for presbyopia and hyperopia patients. By applying radio frequency current at the peripheral regions of cornea, collagen, the most abundant composition of corneal stroma, shrinks due to the heat generated. The shrinkage at the periphery alters the corneal architecture and achieves clearer focus for near vision. In this work we use ...

Multiphoton microscopy has been shown to be a useful tool in studying drug distribution in biological tissues. In addition, fluorescence lifetime imaging provides information about the structure and dynamics of fluorophores based on their fluorescence lifetimes. Fluorescein, a commonly used fluorescent probe, is metabolized within liver cells to ...

Conventionally, the diagnosis of hepatocellular carcinoma (HCC) is performed by qualitative examination of histopathological specimens, which takes times for sample preparation in fixation, section and stain. Our objective is to demonstrate an effective and efficient approach to apply multiphoton microscopy imaging the HCC specimens, with the advantages of ...

The skin is a large and accessible area of the body, offering the possibility to be used as an alternative route for drug delivery. In the last few years strong progress has been made on the developing of nanoparticulate systems for specific applications. The interaction of such small particles with human skin and their possible penetration attracted some interest from ...

Intravital microscopy is a method to study the microcirculation in living tissues. Transillumination, oblique reflected light illumination, continuous and stroboscopic epifluorescence microscopy can be used to visualized specific cells and molecules. Intravital microscopy is further enhanced by the advent of laser scanning.spinning ...

Practical 4Pi microscopy has so far exclusively relied on multiphoton excitation of fluorescence, because the nonlinear suppression of contributions from higher-order sidelobes was mandatory for unambiguous axial superresolution. We show that novel lenses of 74� semiaperture angle enable biological 4Pi microscopy with regular ...

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 ...

Most physiological functions of the kidneys, including the clearance of metabolic waste products, maintenance of body fluid, electrolyte homeostasis, and blood pressure, are achieved by complex interactions between multiple renal cell types and previously inaccessible structures in many organ parts that have been difficult to study. Multiphoton fluorescence ...

Near-IR ultrafast pulse laser and confocal microscope are combined to create a multiphoton multichannel non-linear imaging technique, which allows in situ 3-D characterization of nonfluorescent nanoparticles in biological systems. We observed intense CARS signals generated from various metal oxides due to their high third-order nonlinear susceptibilities (x(3)), which do not ...

The ability to image tissue engineering products without damaging histological procedures is important for the understanding of the dynamics of tissue reorganization and formation. In this work, we test the ability of multiphoton autofluorescence and second harmonic generation microscopy to image engineered tissues following chrondrogenic induction. The ...

Multiphoton-targeted photochemistry was used to selectively inactivate the expression of genes in vertebrate cells. A membrane permeable DNA-associating vital dye, ethidium bromide monoacetate (visible wavelength single photon absorption peak at 530 nm) was used to photosensitize chromosomes in dividing cells. A 100-ps infrared laser beam operating at 1.06 microns was focused ...

We have developed an adaptive optics multiphoton microscope. The multiphoton imaging system combines an ultrafast high-power laser, a scanning unit, a motorized Z-scan device and a photon-counting detector. The adaptive optics module is composed of a Hartmann-Shack wavefront sensor and a MEMS deformable mirror. The impact of compensating the aberrations of ...

We report two novel applications of multiphoton microscopy for pharmacological studies of unstained cardiovascular tissue. First, we show that second harmonic generation (SHG) microscopy of unstained cardiac myocytes can be used to determine the sarcomere length with sub-resolution accuracy, owing to the remarkable contrast of the SHG ...

This manuscript demonstrates a multimodal imaging system which combined multiphoton microscopy (MPM) imaging modality with Fourier domain (FD) optical coherence microscopy (OCM) imaging modality. The system used a single fiber-based femtosecond laser as the light source for both MPM and OCM modality. The femtosecond fiber laser has a ...

We review multiphoton microscopy (MPM) including two-photon autofluorescence (2PAF), second harmonic generation (SHG), third harmonic generation (THG), fluorescence lifetime (FLIM), and coherent anti-Stokes Raman Scattering (CARS) with relevance to clinical applications in ophthalmology. The different imaging modalities are discussed highlighting the ...

We review multiphoton microscopy (MPM) including two-photon autofluorescence (2PAF), second harmonic generation (SHG), third harmonic generation (THG), fluorescence lifetime (FLIM), and coherent anti-Stokes Raman Scattering (CARS) with relevance to clinical applications in ophthalmology. The different imaging modalities are discussed highlighting the ...

Multicolor nonlinear microscopy of living tissue using two- and three-photon-excited intrinsic fluorescence combined with second harmonic generation by supermolecular structures produces images with the resolution and detail of standard histology without the use of exogenous stains. Imaging of intrinsic indicators within tissue, such as nicotinamide adenine dinucleotide, ...

By focusing a pulsed laser beam into a sample, harmonic up-conversion can be generated as well as multi-photon excited fluorescence. Whereas multi-photon excited fluorescence microscopy is well established, the use of multi-harmonic generation for three-dimensional image contrast is very recent. Both techniques can provide similar ...

This Resource Letter provides a guide to the literature on nonlinear optics. Books, journals, and websites are introduced that cover the general subject. Journal articles and websites are cited covering the following topics: second-order nonlinearities in transparent media including second-harmonic generation and optical parametric oscillation, third-order and higher nonlinearities, nonlinear ...

We present a prism-based spectrometer integrated into a multifocal, multiphoton microscope. The multifocal configuration facilitates interrogation of samples under different excitation conditions. Notably, the image plane of the microscope and the image plane of the spectrometer are coincident eliminating the need for an intermediate image plane containing an entrance slit. An ...

Multiphoton imaging of the sub-cellular distribution of photosensitizers can provide important clues to their mechanism of action in tumors. We have used this tool to study distribution and pharmacology of photosensitizers in murine hepatoma tumor cells dosed with various photosensitizers. Upon photoactivation, Rose Bengal yields nearly immediate photolytic release of lysomal ...

In spite of the adverse ageing effects of glycation in vivo, in vitro this process is widely employed to increase stiffness and strength of tissues' and artificial scaffolds'. In-situ optical characterization methods that report on the structures within these materials could clarify the effects of glycation. We employed one-photon fluorescence and multiphoton ...

Images of photoelectron emission from metallic surfaces were obtained with a modified atomic force microscope operating in air. Illumination of the samples was achieved in the near field of a metal-coated microcantilever tip, placed in the beam of a femtosecond pulsed laser that is incident at a grazing angle with respect to the sample surface. Photoelectron currents were measured through the tip ...

Images of photoelectron emission from metallic surfaces were obtained with a modified atomic force microscope operating in air. Illumination of the samples was achieved in the near field of a metal-coated microcantilever tip, placed in the beam of a femtosecond pulsed laser that is incident at a grazing angle with respect to the sample surface. Photoelectron currents were measured through the tip ...

and the microscope slide to obtain the optimum cor- rection collar setting. A fast axial scan through a region on an objective lens. Axial scans across the interface between the specimen volume and the slide are used together intensity and axial resolution in confocal or multiphoton microscopy. This can often be introduced

polynomials, adaptive optics 1. INTRODUCTION Optical microscopes, including the confocal fluorescence.2 0.4 0.6 0.8 1 Signal Figure 1. The principle of operation of a confocal microscope. When a plane (normalised units). 2. CONFOCAL AND MULTI-PHOTON MICROSCOPY The scanning confocal microscope is an important

Microscopy Suite The Laser Scanning Confocal Microscope Instructions for Leica SP2 Microscope #2 By Jon Ekman & Karl Garsha Use of the Multiphoton Laser Scanning Confocal Microscope. Basic operation lamp using the rocker switch on the small box next to the microscope. 6) Turn ON the power

,whichimproveaxialresolutionbyafactorofseven by employing two opposing lenses. Noninvasive axial sections of 80�160 nm thickness deliver more faithful 3D and multiphoton versions of fluorescence microscopy stand out by their ability to provide fast 3D images [2 on top of each other (axial direction). In this case, they can only be distin- guished if ...

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Histological analysis is the clinical standard for assessing tissue health and the identification of pathological states. Its invasive nature dictates that its use should be minimized without compromising diagnostic accuracy. A promising method for minimally invasive histological analysis is optical biopsy, which provides cross sectional or 3D images without any physical sectionings. Optical ...

The purpose of this study is to assess the possible application of multiphoton fluorescence and second harmonic generation (SHG) microscopy for imaging the structural features of keratoconus cornea and to evaluate its potential as being a clinical in vivo monitoring technique. Using the near-infrared excitation source from a titanium-sapphire laser pumped ...

Multiphoton microscopy (MPM) is a high resolution (sub-?m) 3D optical imaging technique that has seen widespread use for microscopy at moderate depth within biological tissue (~1 mm). MPM combined with Fluorescence lifetime imaging (FLIM) and Fluorescent resonant energy transfer (FRET) provides the ability to image protein-protein ...

Spectral and multiphoton imaging is the preferred approach for non-invasive study allowing deeper penetration to image molecular processes in living cells. But currently available fluorescence microscopic techniques based on fluorescence intensity, such as confocal or multiphoton excitation, cannot provide detailed quantitative information about the ...

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 ...

Near infrared (NIR) femtosecond laser scanning microscopy represents a novel and very promising medical diagnostic imaging technology for non-invasive cross-sectional analysis of living biological tissues. In this study multiphoton imaging has been performed to analyze the structural features of extracellular matrix (ECM) components, e.g. collagen and ...

One of the main functions of dermal fibroblasts is the generation of mechanical forces within their surrounding extracellular matrix. Investigating molecules that could modulate fibroblast contraction and act as potent anti aging ingredients requires the development of three-dimensional in situ imaging methodologies for dermal substitute analysis. Here we use multiphoton ...

Dermal fibroblasts are responsible for the generation of mechanical forces within their surrounding extracellular matrix and can be potentially targeted by anti-aging ingredients. Investigation of the modulation of fibroblast contraction by these ingredients requires the implementation of three-dimensional in situ imaging methodologies. We use multiphoton ...

Graft outcomes after kidney transplantation continue to be adversely affected by ischemia-reperfusion injury and rejection. High-resolution, real-time imaging of the transplanted kidney could shed valuable insights into these dynamic processes, but such methodology has not been established. Here we describe a technique for intravital imaging of the transplanted mouse kidney using ...

It is shown that colloidal semiconductor nanocrystals (quantum dots), which are promising fluorophores for multiphoton fluorescence microscopy, exhibit a two-photon absorption saturation effect at moderate powers (not exceeding 10 mW) of femtosecond pumping radiation. An analytical expression for the power of two-photon fluorescence of quantum dots as a ...

Fluorescence resonance energy transfer (FRET) microscopy provides a tool to visualize the protein with high spatial and temporal resolution. In multi-photon FRET microscopy one experiences considerably less photobleaching compared to one-photon excitation since the illumination is the diffraction limited spot and the excitation is ...

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 ...

Super-resolution optical microscopy has attracted great interest among researchers in many fields, especially in biology where the scale of physical structures and molecular processes fall below the diffraction limit of resolution for light. As one of the emerging techniques, structured illumination microscopy can double the resolution by shifting ...