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Reduction of the platelet review rate using the two-dimensional platelet method.

Accurate platelet enumeration is critical for optimal treatment of patients with platelet and bleeding disorders, leukemias, and other neoplasias. The majority of automated hematology analyzers count platelets by size differentiation alone, which may result in falsely elevated platelet counts for samples containing interfering particles such as RBC fragments, microcytes, and cell debris. Most analyzers flag questionable platelet counts, necessitating review of results with confirmation by an alternative method, thus increasing the cost of performing platelet counts and delaying results. We studied the effect of a new platelet analysis method, based on measurement of size and refractive index, on the laboratory review rate for platelet counting. We demonstrated that this method yields higher accuracy for platelet counts in samples with interferences, especially for platelet counts less than 50 x 10(3)/microL (< 50 x 10(9)/L). As a result of the 2-dimensional analysis, the review rate for platelet counts was reduced by 65% in our institution, resulting in substantial savings.

Blood Platelets↗

Scattering of light by stochastically rough particles.

The single particle phase function and the linear polarization for large stochastically deformed spheres have been calculated by Monte Carlo simulation using the geometrical optics approximation. The radius vector of a particle is assumed to obey a bivariate lognormal distribution with three free parameters: mean radius, its standard deviation and the coherence length of the autocorrelation function. All reflections/refractions which include sufficient energy have been included. Real and imaginary parts of the refractive index can be varied without any restrictions. Results and comparisons with some earlier less general theories are presented. Applications of this theory to the photometric properties of atmosphereless bodies and interplanetary dust are discussed.

Astronomical Phenomena↗

Low-coherence interferometer system for the simultaneous measurement of refractive index and thickness.

We have developed a low-coherence interferometer system used for the simultaneous measurement of refractive index n and thickness t of transparent plates. Both the phase index n(p) and group index n(g) can be determined automatically in a wide thickness range of from 10 microm to a few millimeters. Two unique techniques are presented to measure n(p), n(g), and t simultaneously. One allows us to determine n(p), n(g), and t accurately by using a special sample holder, in which the measurement accuracy is 0.3% for the thickness t above 0.1 mm. In the other technique the chromatic dispersion delta n of index is approximately expressed as a function of (n(p) - 1) on the basis of measured values of n(p) and n(g) for a variety of materials, and then the simultaneous measurement is performed with a normal sample holder. In addition, a measurement accuracy of less than 1% is achieved even when the sample is as thin as 20 microm. The measurement time is also 3 min or more.

Calibration↗

Interferometric measurement of the refractive-index distribution in plastic lenses by use of computed tomography.

A nondestructive measurement method that makes possible the measurement of a three-dimensional refractive-index distribution of any shape plastic lens is presented. In this method, a Mach-Zehnder interferometer and shearing interferometer are combined into a single optical system and are used selectively. Interference fringes of a test object that is immersed in matching liquid are detected at various rotation angles. And transmitted wave fronts are calculated with these interference fringes. Finally, the refractive-index distribution is obtained by computed tomography analysis. In addition, accurate control of the matching liquid temperature makes it possible to measure the absolute refractive index of the test object. This system has good performance with a measurement accuracy of 10(-4) or better peak to valley.

Interferometry↗

Measurements of optical properties of high-density media.

Measurements of optical properties (scattering coefficient mu(s), absorption coefficient mu(a), reduced scattering coefficient mu(s)', and asymmetry factor g) have been carried out up to a volume particle concentration of rho = 0.227. The results for mu(s) and mu(s)' show significant deviations from the linear dependence on rho as expected when the independent scattering assumption is fulfilled. The asymmetry factor also changed significantly. In contrast, the dependence of mu(a) remained linear even at the largest concentration investigated. The simple linear dependence of absorption on the chromophore concentration expected from the independent scattering assumption is thus applicable also to spectroscopic measurements of dense media. A comparison with an approximate theoretical model based on the Foldy-Twersky equation is also reported. The model provides a good description of the dependence of mu(s) on particle concentration.

Computer Simulation↗

Interferometric laser diode probing of micrometer- and nanometer-scale materials.

We are developing a method for real-time detection, tracking, and categorization of micrometer- and nanometer-scale particles and materials using light scattered from a swept standing-wave probe. Synchronous, phase-sensitive detection of the weakly scattered optical field is exploited to provide interferometric sensitivity and improve the signal-to-noise ratio, allowing use of low-power laser diode sources and photodiode detectors. To demonstrate the technique, we probe a set of W, C, and Cu microfibers and determine diameters and refractive-index values from a detailed comparison of light-scattering data and a numerical model. We extrapolate these results and discuss the application of laser diode sources and photodiode receivers for the detection and study of nanoscale materials.

Algorithms↗

Phase-slope and group-dispersion calculations in the frequency domain by simple optical low-coherence reflectometry.

We report a new method by which phase slope and group dispersion can be calculated with a simple optical low-coherence reflectometer to quantify physiological conditions. A discrete-time signal processing algorithm based on the first and second derivatives of the phase with respect to wave number was developed from discrete-time Fourier properties. The algorithm avoids the 2pi ambiguity associated with most phase unwrapping. Experimental data collected by use of well-characterized optical materials validated the algorithm, which was minimally sensitive to phase noise. The group dispersion of bovine cornea was measured at various hydrations and was significantly dependent on hydration. The results suggest that group dispersion is an indicator of corneal alterations.

Algorithms↗

Polarization-based compensation of astigmatism.

One approach to aberration compensation of an imaging system is to introduce a suitable phase mask at the aperture plane of an imaging system. We utilize this principle for the compensation of astigmatism. A suitable polarization mask used on the aperture plane together with a polarizer-retarder combination at the input of the imaging system provides the compensating polarization-induced phase steps at different quadrants of the apertures masked by different polarizers. The aberrant phase can be considerably compensated by the proper choice of a polarization mask and suitable selection of the polarization parameters involved. The results presented here bear out our theoretical expectation.

Astigmatism↗

Laser spectral characterization in multiphoton microscopy.

Spectral and temporal characterization is a fundamental task when a tunable Ti:sapphire ultrafast laser system is operated for multiphoton microscopy applications. In the present paper simple procedures are reported that perform laser-peak-emission wavelength and bandwidth measurements without the need of any further instrumentation but a simple and inexpensive diffraction grating, by taking advantage of the confocal microscope imaging capabilities.

Equipment Design↗

Phase-sensitive optical low-coherence reflectometry for the detection of analyte concentrations.

Optical techniques may potentially be used for noninvasive glucose sensing. We investigated the application of phase-sensitive optical low-coherence reflectometry (PS-OLCR) to the measurement of analyte concentrations. The dependence of the PS-OLCR signal on the concentration of various analytes, including aqueous solutions of glucose, calcium chloride, magnesium chloride, sodium chloride, potassium chloride, potassium bicarbonate, urea, bovine serum albumin, and bovine globulin, were determined in clear and turbid media. Obtained results demonstrated (1) a high degree of sensitivity and accuracy of the phase measurements of analyte concentrations with PS-OLCR; (2) a concentration-dependent change in the phase-shift for glucose that is significantly greater than that of other analytes sampled over the same physiological range; and (3) a high submillimolar sensitivity of PS-OLCR for the measurement of glucose concentration. Further exploration of the application of PS-OLCR to the noninvasive, sensitive, and specific monitoring of glucose concentration seems warranted.

Biosensing Techniques↗

Fourier-domain holography in photorefractive quantum-well films.

Fourier-domain holography (FDH) is investigated as a candidate for holographic optical coherence imaging to produce real-time images of structure inside living tissue and turbid media. The effects of spatial filtering, the background intensity distributions, and the role of background noise in determining dynamic range are evaluated for both FDH and image-domain holography (IDH). The grating washout effect in FDH (edge enhancement) is removed by use of a vibrating diffuser that consequently improves the image quality. By comparing holographic images and background images of FDH and IDH we show that FDH provides a higher dynamic range and a higher image quality than IDH for this specific application of imaging diffuse volumetric objects.

Equipment Design↗

Rapid optically based measurements of diameter and length for spherical or rod-shaped bacteria in vivo.

The application in light scattering of the Mueller matrix ratio (S34)/(S11) for determining average particle size is extended to a large size parameter range for spherical or randomly oriented rod-shaped particles such as micro-organisms. It is shown that combining the graph of this ratio with a Coulter counter measurement of particle volume gives results in agreement with microscopic measurements. Thus this combination provides a method to measure particle diameter and width simultaneously in real time for elongated particles such as bacteria, which are measured in vivo with this method. An approximate empirical formula is developed to estimate the motion of the extrema in the graph of the oscillating matrix ratio as size changes occur. This formula is also shown to be consistent with wavelength changes.

Algorithms↗

Spherical beam volume holograms for spectroscopic applications: modeling and implementation.

The spherical beam volume hologram, recorded by a plane wave and a spherical beam, is investigated for spectroscopic applications in detail. It is shown that both the diffracted and the transmitted beam can be used for spectroscopy when the hologram is read with a collimated beam. A new method is introduced and used for analysis of the spherical beam volume hologram that can be extended for analysis of arbitrary holograms. Experimental results are consistent with the theoretical study. It is shown that the spherical beam volume hologram can be used in a compact spectroscopic configuration when the transmitted beam is monitored. Also, on the basis of the properties of the spherical beam hologram, the response of a hologram recorded by a plane wave and an arbitrary pattern is predicted. The information can be used to optimize holographic spectrometer design.

Algorithms↗

Optical frequency-domain reflectometry with a rapid wavelength-scanning superstructure-grating distributed Bragg reflector laser.

Superstructure-grating distributed Bragg reflector lasers are particularly suited for optical frequency-domain reflectometry optical-coherence tomography with wide wavelength tunability and frequency agility. We report theoretical estimates of and experimental results for the data acquisition speed, the observable depth range, the resolution, and the dynamic range of an optical frequency-domain reflectometry system that uses a superstructure-grating distributed Bragg reflector laser whose wavelength can be tuned from 1533 to 1574 nm with a tuning speed of 10 micros/0.1-nm step.

Equipment Design↗

Phase-unwrapping algorithm for images with high noise content based on a local histogram.

We present a robust algorithm of phase unwrapping that was designed for use on phase images with high noise content. We proceed with the algorithm by first identifying regions with continuous phase values placed between fringe boundaries in an image and then phase shifting the regions with respect to one another by multiples of 2pi to unwrap the phase. Image pixels are segmented between interfringe and fringe boundary areas by use of a local histogram of a wrapped phase. The algorithm has been used successfully to unwrap phase images generated in a three-dimensional shape measurement for noninvasive quantification of human skin structure in dermatology, cosmetology, and plastic surgery.

Algorithms↗

Two-dimensional thickness measurements based on internal reflection ellipsometry.

An imaging ellipsometer technique on internal reflection geometry that can measure the thickness distribution of a thin film possessing an assumed refractive index is described. Because a prism is used for the internal reflection geometry, it was theoretically predicted that angular derivation from the normal incidence on the prism surface affects only the psi value by a factor of 0.97 at maximum. Measurements were carried out for an optical system of silica substrate-TiO2 layer-silica layer-protein film-air, with a thin-film array of dried protein as the sample film. Thickness of the protein films was two-dimensionally estimated only from the measured map of the delta value by use of the simulated relationship between the thickness and the delta value. The thickness map obtained was coincident on the whole with the results according to a mechanical scanning. The detection limit was approximately +/- 0.2 nm. These findings validate the optical effect of a high-index additional layer to improve the sensitivity and precision of thickness measurements of the sample film on transparent substrates.

Coated Materials, Biocompatible↗

Properties of a diffused photon-pair density wave in a multiple-scattering medium.

A novel diffused photon-pair density wave (DPPDW) composed of correlated polarized photon pairs at different temporal frequencies and orthogonal linearly polarized states is proposed. A theory of DPPDWs is developed. A DPPDW selected by coherence gating and polarization gating that satisfies the diffusion equation has been verified experimentally. The sensitivity of amplitude and phase detection of the heterodyne signal has been improved by the properties of synchronized detection and common-path propagation of polarized pair photons in a multiple-scattering medium. Both reduced scattering coefficient micro2s' and absorption coefficient micro2alpha of the scattering medium in terms of the measured phase and amplitude of the heterodyne signal have been obtained. The detection sensitivity of micro2s' and micro2alpha and the properties of a DPPDW in a multiple-scattering medium are discussed and analyzed.

Diffusion↗

In vivo scattering measurement of biological tissue by the use of a pyroelectric polymer transducer.

We propose a new method, to our knowledge, for noninvasive scattering measurements of tissues by the use of a pyroelectric polymer transducer, poly(vinylidene fluoride trifluoroethylene) film. In this method, samples are irradiated with nanosecond, low-energy light pulses delivered from an optical fiber, and the pyroelectric signal induced by the diffuse reflectance is measured with a transducer. The signal is then converted into diffuse reflectance by use of a calibration factor. The validity of this method was confirmed by our measuring the scattering coefficient of a white acrylic resin within an accuracy of +/- 15%. We attempted to apply this method to the estimation of scattering coefficients of normal and burned skins in rats in vivo.

Animals↗