This PDF file contains the front matter associated with SPIE Proceedings Volume 9792, including the Title Page, Copyright information, Table of Contents, and Conference Committee listing.

Multidimensional signals such as the spectral images allow us to have deeper insights into the natures of objects. In this paper the spectral imaging techniques that are based on optical interferometry and nonlinear optics are presented. The interferometric imaging technique is based on the unified theory of Van Cittert-Zernike and Wiener-Khintchine theorems and allows us to retrieve a spectral image of an object in the far zone from the 3D spatial coherence function. The retrieval principle is explained using a very simple object. The promising applications to space interferometers for astronomy that are currently in progress will also be briefly touched on. An interesting extension of interferometric spectral imaging is a 3D and spectral imaging technique that records 4D information of objects where the 3D and spectral information is retrieved from the cross-spectral density function of optical field. The 3D imaging is realized via the numerical inverse propagation of the cross-spectral density. A few techniques suggested recently are introduced. The nonlinear optical technique that utilizes stimulated Raman scattering (SRS) for spectral imaging of biomedical targets is presented lastly. The strong signals of SRS permit us to get vibrational information of molecules in the live cell or tissue in real time. The vibrational information of unstained or unlabeled molecules is crucial especially for medical applications. The 3D information due to the optical nonlinearity is also the attractive feature of SRS spectral microscopy.

Surface-enhanced Raman scattering (SERS) continues to strive to gather molecular level information from dynamic biological systems. It is our ongoing effort to utilize the technique for understanding of the biomolecular processes in living systems such as eukaryotic and prokaryotic cells. In this study, the technique is investigated to identify cell death mechanisms in 2D and 3D in vitro cell culture models, which is a very important process in tissue engineering and pharmaceutical applications. Second, in situ biofilm formation monitoring is investigated to understand how microorganisms respond to the environmental stimuli, which inferred information can be used to interfere with biofilm formation and fight against their pathogenic activity.

Raman microscopy is useful for molecular imaging and analysis of biological specimens. Here, we used alkyne containing a carbon-carbon triple bond as a Raman tag for observing small molecules in live cells. Alkyne tags can maintain original properties of target molecules with providing high chemical specificity owing to its distinct peak in a Raman-silent window of biomolecules. For demonstrations, alkyne-tagged thymidine and coenzyme Q analogue in live cells were visualized with high-spatial resolution. We extended the application of alkyne-tag imaging to visualize cell organelles and specific lipid components in artificial monolayer membranes.

The emission intensity of fluorophore molecule may change in presence of strong plasmon field induced by nanoparticles. The enhancement intensity is optimized through selective clustering or functionalization of nanoparticles in closed vicinity of fluorophore. Our study is aimed at understanding the enhancement mechanism of fluorescence intensity in presence of gold nanoparticles to utilize it in molecular sensing and in situ imaging in the microfluidic lab-on-chip device. Related phenomena are studied in situ in a microfluidic channel via fluorescence imaging. Detailed analysis is carried out to understand the possible mechanism of enhancement of fluorescence due to nanoparticles. In the present experimental study we show that SYTO9 fluorescence intensity increased in presence of Au nanoparticles of ~20 nm diameter. The fluorescence intensity is 20 time more compared to that in absence of Au nanoparticles. The enhancement of fluorescence intensity is attributed to the plasmonic resonance of Au nanoparticle at around the fluorescence emission wavelength. Underlying fundamental mechanism via dipole interaction model is explored for quantitative correlation of plasmonic enhancement properties.

Real-time imaging of live cells is quite difficult without the addition of external contrast agents. Various methods for quantitative phase imaging of living cells have been proposed like digital holographic microscopy and diffraction phase microscopy. In this paper, we report theoretical and experimental results of quantitative phase imaging of live yeast cells with nanometric precision using transport of intensity equations (TIE). We demonstrate nanometric depth sensitivity in imaging live yeast cells using this technique. This technique being noninterferometric, does not need any coherent light sources and images can be captured through a regular bright-field microscope. This real-time imaging technique would deliver the depth or 3-D volume information of cells and is highly promising in real-time digital pathology applications, screening of pathogens and staging of diseases like malaria as it does not need any preprocessing of samples.

To provide clinically relevant insights into the device performance of an optical imaging approach to reconstruct the superficial cutaneous micro-circulation (skin angiography), a phantom device with turbid matrix and perfusable micro-vessels is essential. In this work, we describe a novel microfluidic-based device to mimic the micro-vessels and the turbid nature of the epidermis and dermis. This phantom device contains a hollow assay with a diameter of the channels of 50 μm. The hollow assay includes the geometry of the inlet, the river-like assay, and the outlet, which can be perfused by e.g. meta-hemoglobin solution. This imitates the superficial micro-circulation in the skin. The absorption coefficient μ_a and the reduced scattering coefficient μ_s' are adjusted to match those of skin. As an application case, we attempt to reconstruct a 2-D velocity field of the hemoglobin flow in the scattering microfluidic device via the Doppler-mode of an OCT.

Coherent light propagating through turbid media is attenuated due to scattering and absorption. The decrease of the intensity of the coherent light is described by the attenuation coefficient. The measured decay of the coherent light through turbid media with optical coherence tomography (OCT) can be used to reconstruct the attenuation coefficient. Since most of the OCT systems work in the near-infrared region, they are the optical window from 800-1400 nm in tissue. Hence, the most part of the attenuation coefficient is caused due to the scattering. Therefore, deriving the attenuation coefficient is one way to get an approximation of the scattering coefficient which is difficult to access even up to day. Moreover, OCT measurements are one of the few possibilities to derive physical properties with micrometre resolution of the media under investigation.

Multifrequency sensing technique adopting the wide field heterodyne detection technique is demonstrated for interior surface vibration measurements in thick biological tissue. These arrangements allow obtaining not only 3D tomographic images but also various vibration parameters such as spatial amplitude, phase, and frequency, with high temporal and transverse resolutions over a wide field. The axial resolution and the accuracy of vibration amplitude measurement were estimated to be 2.5 μm and 3 nm, respectively. This wide-field tomographic sensing method can be applied for measuring microdynamics of a variety of biological samples, thus contributing to the progress in life sciences research.

Anisotropy factor g, one of the optical properties of biological tissues, is the most important parameter to accurately determine scattering coefficient μs in the inverse Monte Carlo (iMC) simulation. It has been reported that g has wavelength and absorption dependence, however, there are few attempts in order to calculate μs of biological tissue considering the wavelength and absorption dependence of g. In this study, the scattering angular distributions of biological tissue phantoms were measured in order to determine g by using goniometric measurements with three polarization conditions at strongly and weakly absorbing wavelengths of hemoglobin. Then, optical properties, especially, μs were measured by integrating sphere measurements and iMC simulation in order to confirm the influence of measured g on optical properties in comparison of with general value of g (0.9) for soft biological tissue. Consequently, it was found that μs was overestimated at strongly absorbing wavelength, however, μs was underestimated at weakly absorbing wavelength if the g was not considered its wavelength and absorption dependence.

The large number of modes supported by multimode optical fibers potentially allows the transmission of larger amounts of information compared to single mode fibers. However, when pulsed light is transmitted through multimode fibers, the spatio-temporal profile of the incident beam is altered upon propagation, leading to a highly scrambled spatial profile and a broadened temporal duration due to modal and material dispersion. We present a digital phase conjugation method to counter-propagate through a multimode optical fiber only a group of modes of similar propagation constants which interfere constructively at a single location at the other side of the fiber, generating a spatially focused pulse. Since only modes with the same speed are excited, temporal broadening due to modal dispersion is suppressed, preserving the ultrashort duration of the propagating pulse. Using this technique, we experimentally demonstrate the transmission of focused pulses of 500 fs through a 30 cm length, 200 micrometer core step-index multimode fiber. Additionally, using a graded-index fiber, which allows the propagation of a larger number of modes of the same speed than a graded index fiber (hence a better focusing capability), we have been able to deliver and scan high-intensity focused femtosecond pulses. Moreover, based on the described principle, we demonstrate for the first time two-photon excitation imaging through a multimode optical fiber.

Digital holographic microscope (DHM) is an emerging quantitative phase imaging technique with unique imaging scales and resolutions leading to multitude of applications. DHM is promising as a novel investigational and applied tool for cell imaging, studying the morphology and real time dynamics of cells and a number of related applications. The use of numerical propagation and computational digital optics offer unique flexibility to tune the depth of focus, and compensate for image aberrations. In this work, we report imaging the dynamics of cell division in E.coli and yeast cells using a DHM platform. We demonstrate 3-D and depth imaging as well as reconstruction of phase profiles of E.coli and yeast cells using the system. We record a digital hologram of E.coli and yeast cells and reconstruct the image using Fresnel propagation algorithm. We also use aberration compensation algorithms for correcting the aberrations that are introduced by the microscope objective in the object path using linear least square fitting techniques. This work demonstrates the strong potential of a DHM platform in 3-D live cell imaging, fast clinical quantifications and pathological applications.

Development of imaging, sensing, and characterization of cells at Research Center for Applied Sciences (RCAS) of Academia Sinica in Taiwan is progressing rapidly. The research on advanced lattice light sheet microscopy for temporal visualization of cells in three dimensions at sub-cellular resolution shows novel imaging results. Label-free observation on filopodial dynamics provides a convenient assay on cancer cell motility. The newly-developed software enables us to track the movement of two types of particles through different channels and reconstruct the co-localized tracks. Surface plasmon resonance (SPR) for detecting urinary microRNA for diagnosis of acute kidney injury demonstrates excellent sensitivity. A fully automated and integrated portable reader was constructed as a home-based surveillance system for post-operation hepatocellular carcinoma. New microfluidic cell culture devices for fast and accurate characterizations prove various diagnosis capabilities.

Development of a physically accurate and computationally efficient photon migration model for turbid media is crucial for optical computed tomography such as diffuse optical tomography. For the development, this paper constructs a space-time coupling model of the radiative transport equation with the photon diffusion equation. In the coupling model, a space-time regime of the photon migration is divided into the ballistic and diffusive regimes with the interaction between the both regimes to improve the accuracy of the results and the efficiency of computation. The coupling model provides an accurate description of the photon migration in various turbid media in a wide range of the optical properties, and reduces computational loads when compared with those of full calculation of the RTE.

We experimentally prepared a laser speckle flowgraphy (LSFG) device, and investigated whether it can accurately measure cutaneous blood flow. Cutaneous blood flow was measured in healthy adults using the thermal diffusion method (TDM) and strain-gauge plethysmography (SPG) established as blood flow measurement methods, along with LSFG. The correlation properties between the values measured by LSFG and TDM, and LSFG and SPG, were investigated. Results found that a significant positive correlation is noted between the two conventional methods and LSFG. It is suggested that LSFG is a useful device for cutaneous blood flow evaluation, and that its applications in medical care and esthetic fields are expected.

Functional near-infrared spectroscopy (fNIRS) can non-invasively detect hemodynamic changes associated with cerebral neural activation in human subjects. However, its signal is often affected by changes in the optical characteristics of tissues in the head other than brain. To conduct fNIRS measurements precisely and efficiently, the extraction and realtime monitoring of the cerebral functional component is crucial. We previously developed methods for extracting the cerebral functional component—the multidistance optode arrangement (MD) method and the hemodynamic modality separation (HMS) method. In this study, we implemented these methods in a software used with the fNIRS system OEG- 17APD (Spectratech, Japan), and realized a real-time display of the extracted results. When using this system for human subject experiments, the baselines obtained with the MD and HMS methods were highly stabilized, whereas originally, the fNIRS signal fluctuated significantly when the subject moved. Through a functional experiment with repetitive single-sided hand clasping tasks, the extracted signals showed distinctively higher reproducibility than that obtained in the conventional measurements.

An accurate determination of optical properties of agricultural products is crucial for non-destructive assessment of food quality. For the determination, light intensity is measured at the surface of the product; then, inverse analysis is employed based on a light propagation model such as the radiative transfer equation (RTE). The inverse analysis requires high computational loads because the light intensity is numerically calculated using the model every time the optical properties are changed. For the calculation, we propose an efficient technique by combining a numerical solution with an analytical solution of the RTE, and investigate the validity of the technique in a two-dimensional homogeneous circular medium which is regarded as a light propagation model with optical properties of kiwifruit. The proposed technique can provide accurate results of the light intensity in change of the optical properties, and the accuracy is less dependent on the boundary conditions and source-detector angles. In addition, the technique can reduce computation time compared with that for numerical calculation of the RTE. These results indicate usefulness of the proposed technique for the inverse analysis.

A Shack-Hartmann wavefront sensor (SHWFS) which consists of a microlens array and an image sensor has been used to measure the wavefront aberrations in various fields owing to its advantages such as simple configuration. However, a conventional SHWFS has the finite dynamic range. The dynamic range cannot be expanded without sacrificing the spatial resolution and the sensitivity in a conventional SHWFS. In this study, the SHWFS using a dual microlens array to solve the problem is proposed. In the proposed method, an astigmatic microlens is arranged at the center of a group of 2 x 2 spherical microlenses. A pattern image including spots and linear patterns is obtained at the focal plane by the dual microlens array. The pattern image can be separated into two images as if two microlens array with different diameter were used by discriminating spots from linear patterns with the pattern matching technique. The proposed method enables to expand the dynamic range of an SHWFS by using the separated two images. The performance of the proposed method is confirmed by the numerical simulation for measuring a spherical wave.

Natural molecular dye, anthocyanin, from mangosteen rind (Garcinia mangostana L.) is described here as safe sensitizer for TiO2 particles in photodegradation of organic contaminants in water. The dye is a promising replacement for the more costly and hazardous heavy metal based systems, such as Fe-complexes and Ru-complexes. Anthocyanin was used as a sensitizer in TiO2/anthocyanin photoanode. Those photoanodes was effectively catalyzed the photoelectrodegradation of Rhodamine B contaminant under halogen lamp of 300 watt. The natural dye molecules showed the tendency to degrade under photoelectrodegradation conditions.However, complete mineralization of rhodamine B as well as anthocyanin occurred leaving no traces of organic species in solution. Sensitizer degradation caused deactivation of the supported catalyst on recovery. Such a short coming was overcome by re-treatment of the recovered catalysts with fresh dye. It was noted that the NaCl electrolyte significantly influenced the RB

fNIRS (functional near-Infrared spectroscopy) can measure brain activity non-invasively and has advantages such as low cost and portability. While the conventional fNIRS has used laser light, LED light fNIRS is recently becoming common in use. Using LED for fNIRS, equipment can be more inexpensive and more portable. LED light, however, has a wider illumination spectrum than laser light, which may change crosstalk between the calculated concentration change of oxygenated and deoxygenated hemoglobins. The crosstalk is caused by difference in light path length in the head tissues depending on wavelengths used. We conducted Monte Carlo simulations of photon propagation in the tissue layers of head (scalp, skull, CSF, gray matter, and white matter) to estimate the light path length in each layers. Based on the estimated path lengths, the crosstalk in fNIRS using LED light was calculated. Our results showed that LED light more increases the crosstalk than laser light does when certain combinations of wavelengths were adopted. Even in such cases, the crosstalk increased by using LED light can be effectively suppressed by replacing the value of extinction coefficients used in the hemoglobin calculation to their weighted average over illumination spectrum.

Mueller matrix polarimetric imaging (MMPI) provides a complete characterization of an anisotropic optical medium. Subsequent single value decomposition allows image interpretation in terms of basic optical anisotropies, such as depolarization, diattenuation, and retardance. In this work, healthy in-vivo skin at different anatomical locations of a biological model (Rattus norvegicus) was imaged by the MMPI technique using 532nm coherent illumination. The body parts under study were back, abdomen, tail, and calvaria. Because skin components are randomly distributed and skin thickness depends on its location, polarization measures arise from the average over a single detection element (pixel) and on the number of free optical paths, respectively. Optical anisotropies over the imaged skin indicates, mainly, the presence of components related to the physiology of the explored region. In addition, a MMPI-based comparison between a tumor on the back of one test subject and proximal healthy skin was made. The results show that the single values of optical anisotropies can be helpful in distinguishing different areas of in-vivo skin and also lesions.

Most of skin pathologies, including melanoma and basal/squamous cell carcinoma, are related to alterations in external and internal order. Usually, physicians rely on their empirical expertise to diagnose these ills normally assisted with dermatoscopes. When there exists skin cancer suspicion, a cytology or biopsy is made, but both laboratory tests imply an invasive procedure. In this regard, a number of non-invasive optical techniques have been proposed recently to improve the diagnostic certainty and assist in the early detection of cutaneous cancer. Herein, skin optical properties are derived with a multispectral polarimetric dermatoscope using three different illumination wavelength intervals centered at 470, 530 and 635nm. The optical device consist of two polarizing elements, a quarter-wave plate and a linear polarizer, rotating at a different angular velocity and a CCD array as the photoreceiver. The modulated signal provided by a single pixel in the acquired image sequence is analyzed with the aim of computing the Stokes parameters. Changes in polarization state of selected wavelengths provide information about the presence of skin pigments such as melanin and hemoglobin species as well as collagen structure, among other components. These skin attributes determine the local physiology or pathology. From the results, it is concluded that optical polarimetry will provide additional elements to dermatologists in their diagnostic task.