Two-photon autofluorescence spectroscopy and second-harmonic generation of epithelial tissue

A spectroscopy system is developed for studying the two-photon excited fluorescence (TPEF) and second-harmonic generation (SHG) of epithelial tissue in backscattering geometry. Our findings show that TPEF signals from epithelial and underlying stromal layers exhibit different spectral characteristics, providing information on the biomorphology and biochemistry of tissue. The SHG signal serves as a sensitive indicator of collagen to separate the epithelial layer from underlying stroma. The polarization dependence of the SHG signal reveals a well-ordered orientation of collagen fibers in the stromal layer. The results demonstrate the potential of depth-resolved TPEF and SHG in determining the pathology of epithelial tissue.     

Depth-resolved monitoring of glucose diffusion in tissues by using optical coherence tomography

We demonstrate the capability of the optical coherence tomography (OCT) technique for depth-resolved monitoring and quantifying of glucose diffusion in fibrous tissues (sclera). The depth-resolved and average permeability coefficients of glucose were calculated. We found that the glucose diffusion rate is not uniform throughout the tissue and is increased from approximately 2.39+/-0.73 x 10(-6) cm/s at the epithelial side to 8.63+/-0.27 x 10(-6) cm/s close to the endothelial side of the sclera. Results demonstrated that the OCT technique is capable of depth-resolved monitoring and quantification of glucose diffusion in sclera with a resolution of approximately 40 mum.     

Laser-induced fluorescence imaging of subsurface tissue structures with a volume holographic spatial-spectral imaging system

A three-dimensional imaging system incorporating multiplexed holographic gratings to visualize fluorescence tissue structures is presented. Holographic gratings formed in volume recording materials such as a phenanthrenquinone poly(methyl methacrylate) photopolymer have narrowband angular and spectral transmittance filtering properties that enable obtaining spatial-spectral information within an object. We demonstrate this imaging system's ability to obtain multiple depth-resolved fluorescence images simultaneously.     

Coherent backscattering spectroscopy

Coherent backscattering (CBS) of light in random media has been previously investigated by use of coherent light sources. Here we report a novel method of CBS measurement that combines low spatial coherence, broadband illumination, and spectrally resolved detection. We show that low spatial coherence illumination leads to an anomalously broad CBS peak and a dramatic speckle reduction; the latter is further facilitated by low temporal coherence detection. Thus CBS can be observed in biological tissue and other media that previously were beyond the reach of conventional CBS measurements. We also demonstrate, for the first time to our knowledge, spectroscopic analysis of CBS. CBS spectroscopy may find important applications in probing random media such as biological tissue in which depth-selective measurements are crucial.     

Circular polarization memory effect in low-coherence enhanced backscattering of light

We experimentally study the propagation of circularly polarized light in the subdiffusion regime by exploiting enhanced backscattering [(EBS), also known as coherent backscattering] of light under low spatial coherence illumination. We demonstrate for the first time, to the best of our knowledge, that a circular polarization memory effect exists in EBS over a large range of scatterers' sizes in this regime. We show that low-coherence EBS signals from the helicity preserving and orthogonal helicity channels cross over as the mean free path length of light in media varies, and that the cross point indicates the transition from multiple to double scattering in EBS.     

Non-Scanning Optical Coherence Tomography Using Off-Axis Interferometry with an Angular-Dispersion Imaging Scheme

We present a non-scanning approach for real-time optical coherence tomography (OCT). Our approach is based on an off-axis interferometer that laterally projects the time-of-flight or depth information of the sample onto an image sensor. To facilitate the use of an off-axis interferometer in OCT, an angular-dispersion imaging method has been developed. Theoretical analysis and experimental results are presented to demonstrate that this method is capable of demodulating the interferogram and thus permits a direct inspection of the depth-resolved image. Depth resolution and detection range of the present method are also studied.     

Random Backscattering in the Parabolic Scaling

In this paper we revisit the parabolic approximation for wave propagation in random media by taking into account backscattering. We obtain a system of transport equations for the moments of the components of reflection and transmission operators. In the regime in which forward scattering is strong and backward scattering is weak, we obtain closed form expressions for physically relevant quantities related to the reflected wave, such as the beam width, the spectral width and the mean spatial power profile. In particular, we analyze the enhanced backscattering phenomenon, that is, we show that the mean power reflected from an incident quasi-plane wave has a maximum in the backscattered direction. This enhancement can be observed in a small cone around the backscattered direction and we compute the enhancement factor as well as the shape of the enhanced backscattering cone.     

Amplification of enhanced backscattering from a dye-doped polymer bounded by a rough surface

We report the experimental study of the enhanced backscattering from a random rough surface through a laser dye-doped polymer. The sample is a slice of pyrromethene-doped polymer coupled with a two-dimensional rough gold layer with a large slope. When the sample is illuminated with an s-polarized He-Ne laser and pumped by a cw argon-ion laser, amplified backscattering is observed. The enhanced backscattering peak increases sharply and its width narrows for a sample with low dielectric constant |?(2)|.     

Two-dimensional depth-resolved Mueller matrix characterization of biological tissue by optical coherence tomography

We built a polarization-sensitive optical coherence tomographic system and measured the two-dimensional depth-resolved full 4 x 4 Mueller matrix of biological tissue for what is believed to be the first time. The Mueller matrix measurements, which we made by varying the polarization states of the light source and the detector, yielded a complete characterization of the polarization property of the tissue sample. The initial experimental results indicated that this new approach reveals some tissue structures that are not perceptible in standard optical coherence tomography.     

Weak localization effects in the second-harmonic light scattered by random systems of particles

We report rigorous numerical simulations that show the presence of coherent backscattering effects in the second-harmonic generation and scattering of light by random systems of two-dimensional particles. Since the medium composing the particles is assumed to be homogeneous and isotropic, the second-harmonic field is generated mainly by surface effects. For the fundamental frequency, the results present a clear enhanced backscattering peak. In contrast, the second-harmonic scattering patterns present an intensity dip in the backscattering direction.     

Assessing epithelial cell nuclear morphology by using azimuthal light scattering spectroscopy

We describe azimuthal light scattering spectroscopy (phi/LSS), a novel technique for assessing epithelial-cell nuclear morphology. The difference between the spectra measured at azimuthal angles phi = 0 degrees and phi = 90 degrees preferentially isolates the single backscattering contribution due to large (approximately 10 microm) structures such as epithelial cell nuclei by discriminating against scattering from smaller organelles and diffusive background. We demonstrate the feasibility of using phi/LSS for cancer detection by showing that spectra from cancerous colon tissue exhibit significantly greater azimuthal asymmetry than spectra from normal colonic tissues.     

Enhanced and specular backscattering in random media

We study scalar waves probing a heterogeneous medium whose parameters are modeled in terms of a statistically isotropic random field. The medium is terminated by an oblique interface at one end (the bottom) and pressure release type boundary conditions at the other end (the top). The tilt of the bottom interface is relatively small so that the dominant contributions to the wave field are confined to a paraxial tube. This study generalizes the basic formulation in terms of Itô–Schrödinger equations in a one-dimensional deterministic background, describing the macrostructure, to one in which the background is more complicated. It provides the first step toward the analysis of scattered waves in general background media modulated by a random microstructure. We discuss in detail the enhanced backscattering phenomenon or weak localization in this setting, with a tilted interface imbedded in the random medium, and find that the backscattering cone does not depend on the tilt. We also find that the enhanced backscattering phenomenon is not affected by the replacement of a specular interface with a diffusive interface.     

Ehrenfest time and the coherent backscattering off ballistic cavities

If the Ehrenfest time tau(E) of a ballistic cavity is not negligible in comparison to its dwell time tau(D), the weak localization correction to the cavity's transmission is suppressed proportional to exp(-tau(E/tau(D). At the same time, quantum interference enhances the probability of reflection into the mode of incidence by a factor two. This "enhanced backscattering" does not depend on the Ehrenfest time. We show that, in addition to the diagonal enhanced backscattering, there are off-diagonal contributions to coherent backscattering that become relevant if tau(E) > or = tau(D).     

Enhanced backscattering of optical vortex fields

The effect of an incident field with a phase screw dislocation (a so-called optical vortex) on the shape of the enhanced backscattering cone was studied theoretically and demonstrated experimentally. We show that the correlation function of the incident field acts as a filter that modifies the shape of the enhanced backscattering cone. The peak value is reduced, and its width is increased as the topological charge of the phase dislocation increases.     

Spectral-domain optical coherence phase microscopy for quantitative phase-contrast imaging

We describe a novel microscopy technique for quantitative phase-contrast imaging of a transparent specimen. The technique is based on depth-resolved phase information provided by common path spectral-domain optical coherence tomography and can measure minute phase variations caused by changes in refractive index and thickness inside the specimen. We demonstrate subnanometer level path-length sensitivity and present images obtained on reflection from a known phase object and human epithelial cheek cells.     

Internal electron diffraction from atomically ordered subsurface nanostructures in metals

We demonstrate that a part of interface at a subsurface nanocavity in Cu(110) can efficiently induce electron scattering back to the surface even if it is inclined with respect to the surface, if the condition for electron diffraction is fulfilled. This backscattering induces oscillations of electron local density of states at the surface versus electron energy. In agreement with our model calculations, the diffraction is assigned to a specific atomic structure at the interface, and is found to be significantly enhanced by focussing of electron waves for propagation along the [110] direction.     

Extinction coefficient imaging of turbid media using dual structured laser illumination planar imaging

We demonstrate a technique, named dual structured laser illumination planar imaging (SLIPI), capable of acquiring depth-resolved images of the extinction coefficient. This is achieved by first suppressing the multiply scattered light intensity and then measuring the intensity reduction caused by signal attenuation between two laser sheets separated by Δz mm. Unlike other methods also able to measure this quantity, the presented approach is based solely on side-scattering detection. The main advantages of dual SLIPI is that it accounts for multiple scattering, provides two-dimensional information, and can be applied on inhomogeneous media.     

Superenhanced backscattering of light by nanoparticles

We report a physical explanation for the phenomenon wherein the backscattering of light by dielectric particles of sizes between 100 and 1 nm is enhanced by 7-11 orders of magnitude. The phenomenon involves complex composite interactions between a dielectric microsphere and a nanoparticle positioned in close proximity to the microsphere. We provide both analytical and perturbation analyses that show that the enhanced backscattering intensity of a nanoparticle is proportional to the third power of its size parameter. Potential applications of this phenomenon include visible-light detection, characterization, and manipulation of particles as small as a few nanometers.     

Depth-resolved birefringence and differential optical axis orientation measurements with fiber-based polarization-sensitive optical coherence tomography

Conventional polarization-sensitive optical coherence tomography (PS-OCT) can provide depth-resolved Stokes parameter measurements of light reflected from turbid media. A new algorithm that takes into account changes in the optical axis is introduced to provide depth-resolved birefringence and differential optical axis orientation images by use of fiber-based PS-OCT. Quaternion, a convenient mathematical tool, is used to represent an optical element and simplify the algorithm. Experimental results with beef tendon and rabbit tendon and muscle show that this technique has promising potential for imaging the birefringent structure of multiple-layer samples with varying optical axes.