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At least 1,009 records · Page 56Linked to original sources

Stokes polarimetry imaging of rat tail tissue in a turbid medium: degree of linear polarization image maps using incident linearly polarized light.

Illumination with incident linearly polarized light on tissue and polarization state measurements of the remitted light provide a means by which various tissue structures can be differentiated. A rat tail is embedded within a turbid gelatin such that there is a variable depth of medium above it. By varying the incident polarization angle (IPA) of the illuminating linearly polarized light, the geometry, and the orientation angle of the tissue, a series of 2-D degree of linear polarization image maps are created using our Stokes polarimetry imaging technique. The image maps show locations of the polarization-sensitive structures in the rat tail, including soft tissue, intervertebral disks, and tendons. The observed morphologies in the image maps indicate locations where the depolarization of light differs according to the tissue type and underlying layers. The data indicate the importance of varying the IPA, and that tissue dichroism and birefringence affect the degree of linear polarization image maps. Diagnostic information regarding subsurface tissue structures is obtained.

Animals↗

Evanescent-imaging-ellipsometry-based microarray reader.

We describe the development of a label-less ellipsometric imaging microarray reader. The ability of the ellipsometric microarray reader to measure binding of sample to microarray surface is verified using oligonucleotide complementary DNA (cDNA) microarrays. Polarized light illuminates the microarray surface through a glass substrate at an angle beyond the critical angle and changes in the polarization of totally internally reflected light resulting from binding events on the microarray surface are measured. This polarization change is used to measure the thickness of biomolecules bound to the microarray. A prototype ellipsometric imaging microarray reader is constructed and calibrated, and the performance is evaluated with cDNA microarrays. The microarray reader measures changes in refractive index changes as small as 0.0024 and thickness changes as small as 0.28 nm. The optimization of angle of incidence and substrate refractive index necessary to achieve high sensitivity is also described. This ellipsometric technique offers an attractive alternative to fluorescence-microarray readers in some genomic, proteomic, diagnostic, and sensing applications.

Algorithms↗

Phase function measurements on nonspherical scatterers using a two-axis goniometer.

We present a two-axis goniometer for measuring the phase function of scattering media with an angular resolution of about 0.2 deg having 12 decades of dynamic range and covering almost the full solid angle. The setup is evaluated with polystyrene spheres and with perpendicularly and obliquely illuminated thin glass cylinders. The scattering pattern and its intensity distribution are in excellent agreement with analytical theory. A multiple scattering configuration composed of two parallel cylinders is also examined. Finally, the phase function of dentin slabs is measured and its dependence on the dental microstructure is discussed.

Algorithms↗

Large-dynamic-range Shack-Hartmann wavefront sensor for highly aberrated eyes.

A conventional Shack-Hartmann wavefront sensor has a limitation that increasing the dynamic range usually requires sacrificing measurement sensitivity. The prototype large-dynamic-range Shack-Hartmann wavefront sensor presented resolves this problem by using a translatable plate with subapertures placed in conjugate with the lenslet array. Each subaperture is the same size as a lenslet and they are arranged so that they overlap every other lenslet position. Three translations of the plate are required to acquire four images to complete one measurement. This method increases the dynamic range by a factor of two with no subsequent change in measurement sensitivity and sampling resolution of the aberration. The feasibility of the sensor was demonstrated by measuring the higher order aberrations of a custom-made phase plate and human eyes with and without the plate.

Corneal Topography↗

Determination of optical properties of human blood in the spectral range 250 to 1100 nm using Monte Carlo simulations with hematocrit-dependent effective scattering phase functions.

The absorption coefficient mu(a), scattering coefficient mu(s), and anisotropy factor g of diluted and undiluted human blood (hematocrit 0.84 and 42.1%) are determined under flow conditions in the wavelength range 250 to 1100 nm, covering the absorption bands of hemoglobin. These values are obtained by high precision integrating sphere measurements in combination with an optimized inverse Monte Carlo simulation (IMCS). With a new algorithm, appropriate effective phase functions could be evaluated for both blood concentrations using the IMCS. The best results are obtained using the Reynolds-McCormick phase function with the variation factor alpha = 1.2 for hematocrit 0.84%, and alpha = 1.7 for hematocrit 42.1%. The obtained data are compared with the parameters given by the Mie theory. The use of IMCS in combination with selected appropriate effective phase functions make it possible to take into account the nonspherical shape of erythrocytes, the phenomenon of coupled absorption and scattering, and multiple scattering and interference phenomena. It is therefore possible for the first time to obtain reasonable results for the optical behavior of human blood, even at high hematocrit and in high hemoglobin absorption areas. Moreover, the limitations of the Mie theory describing the optical properties of blood can be shown.

Blood Chemical Analysis↗

Feature extraction from light-scatter patterns of Listeria colonies for identification and classification.

Bacterial contamination by Listeria monocytogenes not only puts the public at risk, but also is costly for the food-processing industry. Traditional biochemical methods for pathogen identification require complicated sample preparation for reliable results. Optical scattering technology has been used for identification of bacterial cells in suspension, but with only limited success. Therefore, to improve the efficacy of the identification process using our novel imaging approach, we analyze bacterial colonies grown on solid surfaces. The work presented here demonstrates an application of computer-vision and pattern-recognition techniques to classify scatter patterns formed by Listeria colonies. Bacterial colonies are analyzed with a laser scatterometer. Features of circular scatter patterns formed by bacterial colonies illuminated by laser light are characterized using Zernike moment invariants. Principal component analysis and hierarchical clustering are performed on the results of feature extraction. Classification using linear discriminant analysis, partial least squares, and neural networks is capable of separating different strains of Listeria with a low error rate. The demonstrated system is also able to determine automatically the pathogenicity of bacteria on the basis of colony scatter patterns. We conclude that the obtained results are encouraging, and strongly suggest the feasibility of image-based biodetection systems.

Algorithms↗

Light scattering properties of natural and artificially demineralized dental enamel at 1310 nm.

A fundamental understanding of how near-IR light propagates through sound and carious dental hard tissues is essential for the development of clinically useful optical diagnostic systems, since image contrast is based on changes in the optical properties of these tissues on demineralization. During the caries (decay) process, micropores are formed in the lesion due to partial dissolution of the individual mineral crystals. Such small pores behave as scattering centers, strongly scattering visible and near-IR light. The optical properties of enamel can be quantitatively described by the absorption and scattering coefficients, and the scattering phase function. Our aim is to measure the optical scattering behavior of natural and artificial enamel caries. Near-IR attenuation measurements and angular-resolved goniometer measurements coupled with Monte Carlo simulations are used to determine changes in the scattering coefficient and the scattering anisotropy on demineralization at 1310 nm. An ultra-high resolution digital microradiography system is used to quantify the lesion severity by measurement of the relative mineral loss for comparison with optical scattering measurements. The scattering coefficient increases exponentially with increasing mineral loss. Natural and artificial demineralization increases the scattering coefficient more than two orders of magnitude at 1310 nm, and the scattering is highly forward directed.

Dental Caries↗

Investigation of living pancreas tumor cells by digital holographic microscopy.

Digital holographic microscopy provides new facilities for contactless and marker-free quantitative phase contrast imaging. In this work, a digital holographic microscopy method for the integral refractive index determination of living single cells in cell culture medium is presented. Further, the obtained refractive index information is applied to full field thickness and shape determination of adherent pancreas tumor cells, as well as for analysis of drug-induced dynamic changes of a single cell's cytoskeleton. The results demonstrate that digital holographic microscopy is a quantitative phase contrast technique for living cells under conventional laboratory conditions.

Cell Line, Tumor↗

Total internal reflectance fluorescence reader for selective investigations of cell membranes.

A novel setup for fluorescence measurements of surfaces of biological samples, in particular the plasma membrane of living cells, is described. The method is based on splitting of a laser beam and multiple total internal reflections (TIR) within the bottom of a microtiter plate (cell substrate), such that up to 96 individual samples are illuminated simultaneously by an evanescent electromagnetic field. Main prerequisites are an appropriate thickness and a high transmission of the glass bottom, which is attached to the 96-well cell culture plate by a noncytotoxic adhesive. Glass rods of rectangular cross sections are optically coupled to this bottom for TIR illumination. Fluorescence arising from the plasma membrane of living cells is detected simultaneously from all samples using an integrating charge-coupled device (CCD) camera. The TIR fluorescence reader is validated using cultivated cells incubated with different fluorescent markers, as well as stably transfected cells expressing a fluorescent membrane-associating protein. In addition, particularly with regard to potential pharmaceutical applications, the kinetics of the intracellular translocation of a fluorescent protein kinase c fusion protein upon stimulation of the cells is determined.

Animals↗

Birefringence-based eye fixation monitor with no moving parts.

For the purpose of vision screening, we develop an eye fixation monitor that detects the fovea by its unique radial orientation of birefringent Henle fibers. Polarized near-infrared light is reflected from the foveal area in a bow-tie pattern of polarization states, similar to the Haidinger brush phenomenon. In contrast to previous devices that used scanning systems, this instrument uses no moving parts. It rather utilizes four spots of linearly polarized light-two aligned with the "bright" arms and two aligned with the "dark" arms-of the bow-tie pattern surrounding the fovea. The light reflected from the fundus is imaged onto a quadrant photodetector, whereby the circular polarization component of the polarization state of each reflected patch of light is measured. The signals from the four photodetectors are amplified, digitized, and analyzed. A normalized differential signal is computed to detect central fixation. The algorithm is tested on a computer model, and the apparatus is tested on human subjects. This work demonstrates the feasibility of a fixation monitor with no moving parts.

Adult↗

Interferometric measurements of dynamic changes of tear film.

Tear film stability plays an important role in the quality of vision. We present an interferometric method for assessing the stability of precorneal tear film in real time. A lateral shearing technique is applied as a noninvasive and sensitive method for investigating tear film stability and the smoothness of the tear film surface by quantitative evaluation of the interference fringe pattern. The evaporation of tears and the appearance of tear film break-up between blinks cause changes in the fringe geometry. For quantitative assessment of the fringe smoothness and consequently of the tear film structure, the fast Fourier transform (FFT) is applied. Four parameters are used to quantitatively estimate dynamic changes in the tear film stability of the cornea 20 s after the eye blink. Examples of examined interferograms recorded in patients with healthy eyes, patients suffering from dry-eye syndrome, and patients wearing contact lenses are given. Significant differences between the stability of the tear film in the healthy eye and that in the dry eye and the eye with a contact lens are observed. The favorable influence of artificial tears applied in patients with dry-eye syndrome or contact lenses is also discussed.

Dry Eye Syndromes↗

Collagen solubility correlates with skin optical clearing.

Biomedical optics and photomedicine applications are challenged by the turbidity of most biological tissue systems. Nonreactive, biocompatible chemical agents can induce a reversible reduction in optical scattering of collagenous tissues such as human skin. Herein we show that a chemical agent's tissue optical clearing potential is directly related to its collagen solubility, providing a rational design basis for effective, percutaneous formulations.

Animals↗

Monitoring sarcomere structure changes in whole muscle using diffuse light reflectance.

Normal biomechanical and physiological functions of striated muscles are facilitated by the repeating sarcomere units. Light scattering technique has been used in studying single extracted muscle fibers. However, few studies, if any, have been conducted to investigate the possibility of using optical detection to examine sarcomere structure changes in whole muscles. We conducted a series of experiments to demonstrate that optical scattering properties measured in whole muscle are related to changes in sarcomere structure. These results suggest that photon migration technique has a potential for characterizing in vivo tissue ultrastructure changes in whole muscle.

Animals↗

Low-coherence enhanced backscattering: review of principles and applications for colon cancer screening.

The phenomenon of enhanced backscattering (EBS) of light, also known as coherent backscattering (CBS) of light, has been the object of intensive investigation in nonbiological media over the last two decades. However, there have been only a few attempts to explore EBS for tissue characterization and diagnosis. We have recently made progress in the EBS measurements in tissue by taking advantage of low spatial coherence illumination, which has led us to the development of low-coherence enhanced backscattering (LEBS) spectroscopy. In this work, we review the current state of research on LEBS. After a brief discussion of the basic principle of EBS and LEBS, we present an overview of the unique features of LEBS for tissue characterization, and show that LEBS enables depth-selective spectroscopic assessment of mucosal tissue. Then, we demonstrate the potential of LEBS spectroscopy for predicting the risk of colon carcinogenesis and colonoscopy-free screening for colorectal cancer (CRC).

Animals↗

Investigation of light propagation models to determine the optical properties of tissue from interstitial frequency domain fluence measurements.

Four models, standard diffusion approximation (SDA), single Monte Carlo (SMC), delta-P1, and isotropic similarity (ISM), are developed and evaluated as forward calculation tools in the estimation of tissue optical properties. The inverse calculation uses the ratio of the fluences and phase difference at two locations close to an intensity modulated isotropic source to recover the reduced scattering coefficient mus' and the absorption coefficient mua. Diffusion theory allows recovery of optical properties (OPs) within 5% for media with mus'mua>10. The performance of the delta-P1 model is similar to SDA, with limited enhanced accuracy. The collimation approximation may limit the use of the delta-P1 model for spherical geometry, and/or the fluence may not be accurately calculated by this model. The SMC model is the best, recovering OPs within 10% regardless of the albedo. However, the necessary restriction of the searched OPs space is inconvenient. The performance of ISM is similar to that of diffusion theory for media with mus'mua>10, and better for 1 5.

Algorithms↗

Temperature-dependent optical properties of individual vascular wall components measured by optical coherence tomography.

Optical properties of tissues and tissue components are important parameters in biomedical optics. We report measurements of tissue refractive index n and the attenuation coefficient mu(t) using optical coherence tomography (OCT) of individual vascular wall layers and plaque components. Moreover, since the temperature dependence of optical properties is widely known, we compare measurements at room and body temperatures. A decrease of n and mu(t) is observed in all samples, with the most profound effect on samples with high lipid content. The sample temperature is of influence on the quantitative measurements within OCT images. For extrapolation of ex-vivo experimental results, especially for structures with high lipid content, this effect should be taken into account.

Body Temperature↗

In vivo confocal scanning laser microscopy: comparison of the reflectance and fluorescence mode by imaging human skin.

Optical, noninvasive methods have become efficient in vivo tools in dermatological diagnosis and research. From these promising imaging techniques, only the confocal scanning laser microscopy (CSLM) provides visualization of subsurface skin structures with resolutions similar to those of light microscopy. Skin annexes, as well as cutaneous cells from different epidermal layers, can be distinguished excellently. Currently, two forms of application have been established in dermatological practice: the reflectance mode, predominantly in the clinical field, and the fluorescence mode in dermatological research. Differences in both methods exist in the preparative protocol, in maximum imaging depth and, particularly, in the gain of contrast extraction. The reflectance mode demonstrates naturally occurring tissue components, whereas the fluorescent CSLM achieves contrast by administering fluorescence dye, representing the dynamic distribution pattern of the dye's fluorescent emission. Therefore, the reflectance and fluorescent modes highlight various skin microstructures, providing dissimilar in vivo confocal images of the skin. This permits different predications and information on the state of the tissue. We report the advantages and disadvantages of both optical imaging modes. The comparison was drawn by scanning human skin in vivo. Representative images in varying depths were obtained and analyzed; preparation procedures are shown and discussed.

Humans↗

Ultrasharp light-scattering resonances of structured nanospheres: effects of size-dependent dielectric functions.

Recently, Chen et al. [J. Biomed. Opt. Vol. 10, 024005 (2005)] reported on the concept of multicolor molecular imaging, which uses resonant light-scattering spectroscopy of multilayer nanospheres. They claimed that the resonance scattering peaks of three-layer nanoshells can be designed so that the ultrasharp widths are as narrow as 10 nm. Here we show that such ultrasharp labels cannot be fabricated in reality because the effects of size-dependent dielectric functions result in the five- to tenfold broadening of resonant scattering peaks. Furthermore, contrary to the data of Chen et al., we did not find any significant advantages of three-layer structures, as compared with the usual silica/metal nanoshells.

Computer Simulation↗