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PubMedDate not supplied

Optical scattering, absorption, and polarization of healthy and neovascularized human retinal tissues.

The optical scattering, absorption, and polarization properties of human retinal tissues are investigated for a number of laser wavelengths in the visible range. The indices of refraction of these tissues are determined by applying Brewster's law. The inverse adding doubling method based on the diffusion approximation and radiative transport theory is applied to the measured values of total diffuse transmission, total diffuse reflection, and index of refraction to determine the optical absorption, scattering, and scattering anisotropy coefficients of the intact retinal tissues from healthy and diseased (neovascularized) human eyes. The polarization studies show that the retinal tissues possess significant intrinsic polarization characteristics, that are more pronounced in diseased tissues than in healthy tissues.

Absorption↗
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Optical spectroscopy noninvasively monitors response of organelles to cellular stress.

Fast and noninvasive detection of cellular stress is extremely useful for fundamental research and practical applications in medicine and biology. We discovered that light scattering spectroscopy enables us to monitor the transformations in cellular organelles under thermal stress. At the temperatures triggering expression of heat shock proteins, the refractive index of mitochondria increase within 1 min after the onset of heating, indicating enhanced metabolic activity. At higher temperatures and longer exposures, the organelles increase in size. This technique provides an insight into metabolic processes within organelles larger than 50 nm without exogenous staining and opens doors for noninvasive real-time assessment of cellular stress.

Animals↗
PubMedDate not supplied

Parallel dual-color fluorescence cross-correlation spectroscopy using diffractive optical elements.

Dual-color cross-correlation spectroscopy allows the detection and quantification of labeled biomolecules at ultra-low concentrations, whereby the sensitivity of the assay correlates with the measurement time. We now describe a parallel multifocal dual-color spectroscopic configuration employing multiple avalanche photodiodes and hardware correlators. Cross-correlation curves are obtained from several dual-color excitation foci simultaneously. Multifocal dual-color excitation is achieved by splitting each of two laser beams (488 and 633 nm) into four sub-beams with the help of two 2x2 fan-out diffractive optical elements (DOEs), and subsequent superposition of the two sets of four foci. The fluorescence emission from double-labeled biomolecules is detected by two 2x2 fiber arrays.

Equipment Design↗
PubMedDate not supplied

Determination of visible near-IR absorption coefficients of mammalian fat using time- and spatially resolved diffuse reflectance and transmission spectroscopy.

In-vivo optical spectroscopy and the determination of tissue absorption and scattering properties have a central role in the development of novel optical diagnostic and therapeutic modalities in medicine. A number of techniques are available for the optical characterization of tissue in the visible near-IR region of the spectrum. An important consideration for many of these techniques is the reliability of the absorption spectrum of the various constituents of tissue. The availability of accurate absorption spectra in the range 600 to 1100 nm may allow for the determination of the concentration of key tissue constituents such as oxy- and deoxy-hemoglobin, water, and lipids. The objective of the current study is the determination of a reliable absorption spectrum of lipid(s) that can be used for component analysis of in-vivo spectra. We report the absorption spectrum of a clear purified oil obtained from pig lard. In the liquid phase above 36 degrees C, the oil is transparent and thus suitable for collimated transmission measurements. At room temperature, the oil is a solid grease that is highly scattering. The absorption and scattering properties in this solid phase are measured using time- and spatially resolved diffuse reflectance spectroscopy. Using these three independent measurement techniques, we have determined an accurate estimate for the absorption spectrum of mammalian fat.

Adipose Tissue↗
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Light scattering from rat nervous system measured intraoperatively by near-infrared reflectance spectroscopy.

Our goal is to quantify scattering properties of near-IR light in the rat spinal cord region and to differentiate healthy and demyelinated peripheral nerves intraoperatively based on differential light scattering. For the rat spinal cord, optical reflectance is measured from the spinal cord surface at spatial intervals of 1 mm using a needle probe. Data are acquired from left and right lumbar regions of the animals as well as on the central blood vessels. The reduced scattering coefficient mu(s)' is found to be higher (34.2+/-2.1 cm(-1)) in the lumbar regions of the spinal cord than on the central blood vessel (19.9+/-1.0 cm(-1)). This methodology is extended to detect differences in the rat sciatic nerves following left L4 spinal nerve ligation. The reflectance is taken at the same five regions at postoperative days 1, 4, 7, and 14. Significant differences are seen in both the spectral slope and mu(s)' values on postoperative days 4, 7, and 14, indicating that either of the two quantities could be used as a marker for demyelination. We prove the usefulness of the technique, which may have a possible clinical application for minimally invasive, intraoperative diagnosis and monitoring of demyelination diseases, such as multiple sclerosis in the central nervous system or degeneration of the peripheral nervous system.

Animals↗
PubMedDate not supplied

Optical imaging of turbid media using independent component analysis: theory and simulation.

A new imaging approach for 3-D localization and characterization of objects in a turbid medium using independent component analysis (ICA) from information theory is developed and demonstrated using simulated data. This approach uses a multisource and multidetector signal acquisition scheme. ICA of the perturbations in the spatial intensity distribution measured on the medium boundary sorts out the embedded objects. The locations and optical characteristics of the embedded objects are obtained from a Green's function analysis based on any appropriate model for light propagation in the background medium. This approach is shown to locate and characterize absorptive and scattering inhomogeneities within highly scattering medium to a high degree of accuracy. In particular, we show this approach can discriminate between absorptive and scattering inhomogeneities, and can locate and characterize complex inhomogeneities, which are both absorptive and scattering. The influence of noise and uncertainty in background absorption or scattering on the performance of this approach is investigated.

Biopolymers↗
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Polarized laser Doppler perfusion imaging--reduction of movement-induced artifacts.

Laser Doppler perfusion imaging (LDPI) enables superficial tissue perfusion assessment, but is sensitive to tissue motion not related to blood cells. The aim was to investigate if a polarization technique could reduce movement-induced artifacts. A linearly polarized laser and a cross-polarized filter, placed in front of the detectors, were used to block specular reflection. Measurements were performed with, and without, the polarization filter, at a single site during horizontal and vertical movement of skin tissue (index finger, twelve subjects, n = 112) and of a flow model (n = 432), with varying surface structures. Measurements were repeated during different flow conditions and at increased skin specular reflection. Statistical analysis was performed using ANOVA models. The perfusion signal was lower (p < 0.001, skin and p < 0.05, flow model) using the polarization filter, due to movement artifact reduction. No significant influence from surface structure was found when using the polarization filter. Movement artifacts were lower (p < 0.05) in the vertical movement direction, however, depending on flow conditions for skin measurements. Increased skin specular reflection gave rise to large movement artifacts without the polarization filter. In conclusion, the polarized LDPI technique reduces movement artifacts and is particularly appropriate when assessing, e.g., ulcers and burns, where specular reflection is high.

Adult↗
PubMedDate not supplied

Finite element analysis applied to cornea reshaping.

A 2-D finite element model of the cornea is developed to simulate corneal reshaping and the resulting deformation induced by refractive surgery. In the numerical simulations, linear and nonlinear elastic models are applied when stiffness inhomogeneities varying with depth are considered. Multiple simulations are created that employ different geometric configurations for the removal of the corneal tissue. Side-by-side comparisons of the different constitutive laws are also performed. To facilitate the comparison, the material property constants are identified from the same experimental data, which are obtained from mechanical tests on corneal strips and membrane inflation experiments. We then validate the resulting models by comparing computed refractive power changes with clinical results. Tissue deformations created by simulated corneal tissue removal using finite elements are consistent with clinically observed postsurgical results. The model developed provides a much more predictable refractive outcome when the stiffness inhomogeneities of the cornea and nonlinearities of the deformations are included in the simulations. Finite element analysis is a useful tool for modeling surgical effects on the cornea and developing a better understanding of the biomechanics of the cornea. The creation of patient-specific simulations would allow surgical outcomes to be predicted based on individualized finite element models.

Computer Simulation↗
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Transcranial measurement of diffuse light reflectance from cold-injured brains in rats.

Diagnosis of brain edema is important to manage severe head injuries and diseases, but there is no method of noninvasive bedside monitoring of brain edema. We speculate that brain edema changes the optical properties of brain tissue. To verify this speculation, we perform transcranial measurement of diffuse light reflectance from cold-injured brains in rats. To induce edema, a liquid nitrogen-cooled copper probe of 3 mm diameter was applied to the right parietal bone for 60 s under anesthesia. The rat skull was irradiated with 633-nm HeNe laser light and 532-nm Nd:YVO4 laser light, which were used to investigate changes in light scattering and blood flow, respectively. Reflectance intensity was measured as functions of time and position on the skull. We found that reflectance intensities were significantly decreased around the cold injury at both 633 and 532 nm, while reflectance was increased in the cold injury at 532 nm, indicating that cerebral blood volume was decreased in the cold injury. Under the condition of decreased cerebral blood volume, the decrease in reflectance intensity around the injury suggests that the scattering coefficient of brain tissue was reduced due to edema formation in this area.

Animals↗
PubMedDate not supplied

Simulating the response of terahertz radiation to basal cell carcinoma using ex vivo spectroscopy measurements.

Studies of basal cell carcinoma using terahertz pulsed imaging have revealed a significant difference between regions of tumor and healthy tissue. These differences are manifested in the reflected pulse due to what is thought to be changes in refractive index and absorption. We present measurements of the refractive index and absorption coefficient of excised normal tissue and basal cell carcinoma using terahertz (THz) transmission spectroscopy. We extract Debye parameters from these data and enter them into a finite difference time domain simulation to predict the shape of the waveforms reflected off the normal tissue and basal cell carcinoma and compare them with published in vivo data. Simulating the interaction of terahertz radiation with normal and cancerous tissue is a key step toward understanding the origin of contrast in terahertz images of skin cancer.

Basal Cell Carcinoma↗
PubMedDate not supplied

Novel algorithm of processing optical coherence tomography images for differentiation of biological tissue pathologies.

A numerical algorithm based on a small-angle approximation of the radiative transfer equation (RTE) is developed to reconstruct scattering characteristics of biological tissues from optical coherence tomography (OCT) images. According to the algorithm, biological tissue is considered to be a layered random medium with a set of scattering parameters in each layer: total scattering coefficient, variance of a small-angle scattering phase function, and probability of backscattering, which fully describe the OCT signal behavior versus probing depth. The reconstruction of the scattering parameters is performed by their variation to fit the experimental OCT signal by the theoretical one using a time-saving genetic algorithm. The proposed reconstruction procedure is tested on model media with known scattering parameters. The possibility to estimate scattering parameters from OCT images is studied for various regimes of OCT signal decay. The developed algorithm is applied to reconstruct optical characteristics of epithelium and stroma for normal cervical tissue and its pathologies, and the potential to distinguish between the types of pathological changes in epithelial tissue by its OCT images is demonstrated.

Algorithms↗
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Determination of the complex refractive index of highly concentrated hemoglobin solutions using transmittance and reflectance measurements.

The complex refractive index of highly concentrated hemoglobin solutions as they appear in red blood cells are determined in the wavelength range of 250 to 1100 nm using transmittance and Fresnel reflectance measurements. The determined real parts of the refractive indices are on average 0.02 units higher than the values found in the literature. The wavelength dependence of the measured data in the UV region differs from the calculated data using the Kramers-Kronig relation.

Blood Chemical Analysis↗
PubMedDate not supplied

Polarization-correlation mapping of biological tissue coherent images.

We investigate the statistical polarization parameters of biological tissue histological section images with different morphological structure. First we outline the results of polarization coordinate mapping and analysis of the statistics of the first to fourth orders of biological tissue image polarization azimuth and ellipticities. Second, we study the statistics of the first to fourth orders of coordinate distributions of the complex degree of mutual polarization (CDMP) of biological tissue images. Finally, we consider the diagnostic possibilities of investigating 2-D distributions of CDMP of images that correspond to physiologically normal and degeneratively and/or dystrophycally changed biological tissues that are being analyzed.

Animals↗
PubMedDate not supplied

Complex degree of mutual polarization of biological tissue coherent images for the diagnostics of their physiological state.

This is the first attempt devoted to the 1st-4th order statistic analysis of the complex degree of mutual polarization of biological tissues coherent images. The ensemble of diagnostic criteria (skewness and kurtosis of two-dimensional distributions of complex degree of mutual polarization) of pathological changes of muscular and connective (skin derma) tissues is defined.

Algorithms↗
PubMedDate not supplied

Corneal polarimetry after LASIK refractive surgery.

Imaging polarimetry provides spatially resolved information on the polarization properties of a system. In the case of the living human eye, polarization could be related to the corneal biomechanical properties, which vary from the normal state as a result of surgery or pathologies. We have used an aberro-polariscope, which we recently developed, to determine and to compare the spatially resolved maps of polarization parameters across the pupil between normal healthy and post-LASIK eyes. The depolarization distribution is not uniform across the pupil, with post-surgery eyes presenting larger levels of depolarization. While retardation increases along the radius in normal eyes, this pattern becomes irregular after LASIK refractive surgery. The maps of slow axis also differ in normal and post-surgery eyes, with a larger disorder in post-LASIK eyes. Since these changes in polarization indicate subtle structural modifications of the cornea, this approach can be useful in a clinical environment to follow the biomechanical and optical changes of the cornea after refractive surgery or for the early diagnosis of different corneal pathologies.

Adult↗
PubMedDate not supplied

Direct measurement of the evanescent field profile produced by objective-based total internal reflection fluorescence.

Total internal reflection fluorescence (TIRF) microscopy produces a thin excitation field (the evanescent field) that nominally decays exponentially. This field is ideal for selective excitation of fluorophores near the coverslip/sample interface. We present an experimental method, where the depth and axial profile of the evanescent field can be measured directly by microscopic observation of low refractive index fluorescently labeled spherical beads in an index-matched solution. To demonstrate the technique, through-the-objective TIRF is set up with laser excitation. In this configuration, the axial profile of the evanescent field created by either a 1.45-numerical aperture (NA) or a 1.65-NA objective fits well to a double exponential. At the coverslip/sample interface, about 90% of the evanescent field is represented by an exponential with a decay rate consistent with that expected for a theoretical evanescent field; the remaining 10% of the field is represented by an exponential with a much longer decay constant and is identified as scattering. The approach presented here is particularly useful for investigating the quality and axial profile of the evanescent field in both laser-based and mercury arc-based through-the-objective TIRF systems where a significant amount of light scattering can occur in the illumination optics.

Image Interpretation, Computer-Assisted↗
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Remineralization of in vitro dental caries assessed with polarization-sensitive optical coherence tomography.

Polarization-sensitive optical coherence tomography (PS-OCT) is potentially useful for imaging the nonsurgical remineralization of dental enamel. This study uses an all-fiber-based PS-OCT system operating at 1310 nm to image demineralized and fluoride-enhanced remineralized artificial lesions. PS-OCT images of lesions before and after remineralization are compared with the relative mineral loss DeltaZ (%vol x microm), obtained from high resolution digital microradiography (DM), and chemical composition changes by infrared spectroscopy. Severe early artificial caries show a significant increase in perpendicular-axis integrated reflectivity after remineralization. After sectioning the samples, DM demonstrates that the lesions remineralized with new mineral and the lesion surface zone show significant restoration of mineral volume. PS-OCT and DM both do not show a major change in lesion depth. For less severe artificial caries, the perpendicular-axis image resolves the scattering and depolarization of an outer growth layer after remineralization. This outer layer has a mineral volume close to that of sound enamel, and spectroscopic analysis indicates that the layer is a highly crystalline phase of apatite, without carbonate substitutions that increase the solubility of sound enamel. This study determines that PS-OCT can image the effects of fluoride-enhanced remineralization of mild and severe early artificial in vitro caries.

Bone Density↗
PubMedDate not supplied

Structural investigations of human hairs by spectrally resolved ellipsometry.

Human hair is a biological layered system composed of two major layers, the cortex and the cuticle. We show spectrally resolved ellipsometry measurements of the ellipsometric parameters Psi and Delta of single human hairs. The spectra reflect the layered nature of hair and the optical anisotropy of the hair's structure. In addition, measurements on strands of human hair show a high reproducibility of the ellipsometric parameters for different hair fiber bundles from the same person. Based on the measurements, we describe a dielectric model of hair that explains the spectra in terms of the dielectric properties of the major parts of hair and their associated layer thicknesses. In addition, surface roughness effects modeled by a roughness layer with a complex refractive index given by an effective medium approach can be seen to have a significant effect on the measurements. We derive values for the parameters of the cuticle surface roughness layer of the thickness d(ACu)=273 to 360 nm and the air inclusion fA=0.6 to 5.7%.

Algorithms↗