Frequency-domain Photon Migration Measurements Research Articles

Vertical-cavity surface-emitting laser sources for gigahertz-bandwidth, multiwavelength frequency-domain photon migration.

.Frequency-domain photon migration (FDPM) uses modulated laser light to measure the bulk optical properties of turbid media and is increasingly applied for noninvasive functional medical imaging in the near-infrared. Although semiconductor edge-emitting laser diodes have been traditionally used as miniature light sources for this application, we show that vertical-cavity surface-emitting lasers (VCSELs) exhibit output power and modulation performance characteristics suitable for FDPM measurements of tissue optical properties at modulation frequencies exceeding 1 GHz. We also show that an array of multiple VCSEL devices can be coherently modulated at frequencies suitable for FDPM and can improve optical power. In addition, their small size and simple packaging make them an attractive choice as components in wearable sensors and clinical FDPM-based optical spectroscopy systems. We demonstrate the benefits of VCSEL technology by fabricating and testing a unique, compact VCSEL-based optical probe with an integrated avalanche photodiode. We demonstrate sensitivity of the VCSEL-based probe to subcutaneous tissue hemodynamics that was induced during an arterial cuff occlusion of the upper arm in a human subject.

Frequency domain photon migration measurements of dense monodisperse charged lattices and analysis using solutions of Ornstein Zernike equations

Isotropic scattering coefficient measurements were made of monodisperse polystyrene lattices of two different diameters of 144nm and 223nm and at volume fractions ranging from 0.15 to 0.22, using frequency domain photon migration measurements at wavelengths of 660, 685, 785 and 828nm. The isotropic scattering coefficient measurements were shown to be sensitive to the changing ionic strength (0.5–4mM, NaCl equiv.) of the dispersions exhibiting hindered scattering owing to structure at the lowest ionic strength values. Monte Carlo simulations and numerical solution of the Ornstein Zernike equations were used to compute isotropic scattering coefficients for comparison to measured values. The interaction potential was modeled as a hard sphere Yukawa potential and the Hypernetted Chain closure was used to solve the OZ equation. Effective particle charges were found after renormalization of the bare particle charge and used to predict the isotropic scattering coefficient. The model data were found to follow similar trends as experimental measurements. The refractive index of the particles has found to be an important factor for predicting experimental isotropic scattering coefficient values.

Sensitivity and Depth Penetration of Continuous Wave Versus Frequency-domain Photon Migration Near-infrared Fluorescence Contrast-enhanced Imaging ¶

The development of near-infrared fluorescent contrast agents and imaging techniques depends on the deep penetration of excitation light through several centimeters of tissue and the sensitive collection of the re-emitted fluorescence. In this contribution, the sensitivity and depth penetration of various fluorescence-enhanced imaging studies is surveyed and compared with current studies using continuous wave (CW) and frequency-domain photon migration (FDPM) measurements with planar wave illumination of modulated excitation light at 100 MHz and area collection of reemitted fluorescent light using a previously developed modulated intensified charge-coupled device camera system. Fluorescence was generated from nanomolar to micromolar solutions of indocyanine green (ICG) in a 100 μL volume submerged at 1–4 cm depths in a 1% Liposyn® solution to mimic tissue scattering properties. Enhanced depth penetration and sensitivity are achieved with optimal filter rejection of excitation light, and FDPM rejection of background light is not achieved using CW methods. We show the ability to detect as few as 100 fmol of ICG from area illumination of 785 nm light (5.5 mW/cm2) and FDPM area collection of 830 nm fluorescent light generated from 3 cm below the phantom surface. The lowered noise floor of FDPM measurements enables greater sensitivity and penetration depth than comparable CW measurements.

Light diffusion model for determination of optical properties of rectangular parallelepiped highly scattering media

A method to determine the absorption and reduced scattering coefficients of rectangular parallelepiped highly scattering media from frequency-domain photon migration measurements is presented. An analytical model for photon diffusion propagation in the rectangular parallelepiped media is established using the method of images and extrapolated boundary conditions. This present technique has simplicity, accuracy, and rapid computability as compared with the Monte Carlo or finite element methods. The theoretical predictions are verified with experimental measurements using a white polyacetal resin, and the errors introduced by using the slab geometry for the optical property determination are identified.

Monte Carlo simulation of time‐dependent, transport‐limited fluorescent boundary measurements in frequency domain

Recently, we have presented and experimentally validated a unique numerical solver of the coupled radiative transfer equations (RTEs) for rapidly computing time-dependent excitation and fluorescent light propagation in small animal tomography. Herein, we present a time-dependent Monte Carlo algorithm to validate the forward RTE solver and investigate the impact of physical parameters upon transport-limited measurements in order to best direct the development of the RTE solver for optical tomography. Experimentally, the Monte Carlo simulations for both transport-limited and diffusion-limited propagations are validated using frequency domain photon migration measurements for 1.0%, 0.5%, and 0.2% intralipid solutions containing 1 microM indocyanine green in a 49 cm3 cylindrical phantom corresponding to the small volume employed in small animal tomography. The comparisons between Monte Carlo simulations and the numerical solutions result in mean percent error in amplitude and the phase shift less than 5.0% and 0.7 degrees, respectively, at excitation and emission wavelengths for varying anisotropic factors, lifetimes, and modulation frequencies. Monte Carlo simulations indicate that the accuracy of the forward model is enhanced using (i) suitable source models of photon delivery, (ii) accurate anisotropic factors, and (iii) accurate acceptance angles of collected photons. Monte Carlo simulations also show that the accuracy of the diffusion approximation in the small phantom depends upon (i) the ratio d(phantom)/l(tr), where d(phantom) is the phantom diameter and l(tr) is the transport mean free path; and (ii) the anisotropic factor of the medium. The Monte Carlo simulations validates and guides the future development of an appropriate RTE solver for deployment in small animal optical tomography.

Influence of excitation light rejection on forward model mismatch in optical tomography

Fluorescence enhanced tomography for molecular imaging requires low background for detection and accurate image reconstruction. In this contribution, we show that excitation light leakage is responsible for elevated background and can be minimized with the use of gradient index (GRIN) lenses when using fibre optics to collect propagated fluorescence light from tissue or other biological media. We show that the model mismatch between frequency-domain photon migration (FDPM) measurements and the diffusion approximation prediction is decreased when GRIN lenses are placed prior to the interference filters to provide efficient excitation light rejection. Furthermore, model mismatch is correlated to the degree of excitation light leakage. This work demonstrates the importance of proper light filtering when designing fluorescence optical imaging and tomography.

Compression-induced changes in the physiological state of the breast as observed through frequency domain photon migration measurements

We use optical spectroscopy to characterize the influence of mammographic-like compression on the physiology of the breast. We note a reduction in total hemoglobin content, tissue oxygen saturation, and optical scattering under compression. We also note a hyperemic effect during repeated compression cycles. By modeling the time course of the tissue oxygen saturation, we are able to obtain estimates for the volumetric blood flow (1.64+/-0.6 mL/100 mL/min) and the oxygen consumption (1.97+/-0.6 micromol/100 mL/min) of compressed breast tissue. These values are comparable to estimates obtained from previously published positron emission tomography (PET) measurements. We conclude that compression-induced changes in breast physiological properties are significant and should be accounted for when performing optical breast imaging. Additionally, the dynamic characteristics of the changes in breast physiological parameters, together with the ability to probe the tissue metabolic state, may prove useful for breast cancer detection.

Fluorescence-enhanced optical tomography of a large tissue phantom using point illumination geometries

We demonstrate fluorescence-enhanced optical imaging of single and multiple fluorescent targets within a large (approximately 1081 cm3) phantom using frequency-domain photon migration measurements of fluorescence collected at individual points in response to illumination of excitation light at individual points on the boundary. The tissue phantom was filled with a 1% lipid solution with and without 0.01 microM Indocyanine Green (ICG) and targets consisted of vials filled with the 1% lipid containing 1-2.5 microM ICG. Measurements were acquired using a modulated intensified CCD imaging system under different experimental conditions. For 3-D image reconstruction, the gradient-based penalty modified barrier function (PMBF) method with simple bounds constrained truncated Newton with trust region method (CONTN) was used. Targets of 0.5, 0.6, and 1.0 cm3 at depths of 1.4-2.8 cm from the phantom surface were tomographically reconstructed. This work demonstrates the practicality of fluorescence-enhanced tomography in clinically relevant volumes.

Sampling tissue volumes using frequency-domain photon migration.

We demonstrate the use of Monte Carlo simulations to generate photon scattering density functions (PSDFs) that represent the tissue volume sampled by steady-state and frequency-domain photon migration. We use these results to illustrate how scaling laws can be developed to determine the mean sampling depth of the multiply scattered photons detected by photon migration methods that remain valid outside the bounds of the standard diffusion approximation, i.e., at small source-detector separations and in media where the optical absorption is significant relative to scattering. Using both the PSDF computation and the newly formulated scaling laws, we focus on a comprehensive description of the effects of source modulation frequency, optical absorption, and source-detector separation on the depth of the sampled tissue volume as well as the sensitivity of frequency-domain photon migration measurements to the presence of a localized absorption heterogeneity.

Measurement of Low-Dose Active Pharmaceutical Ingredient in a Pharmaceutical Blend Using Frequency-Domain Photon Migration

Frequency-domain photon migration (FDPM) measurements of time-dependent light propagation are conducted to provide the powder absorbance for quantitative prediction of terazosin as the active pharmaceutical ingredient (API) in a low-dose (0.72 wt %) oral tablet formulation. Calibration of the FDPM-derived powder absorbance at discrete wavelengths of 514, 650, 687, and 785nm was performed for API contents ranging between 0 and 1.5 wt % in mixtures showing maximum sensitivity at 650nm. The relative standard deviation (RSD) of FDPM absorption coefficient measurement at 650nm in a well-mixed 1.08 wt % terazosin blend was <1.6%, of which no more than 0.12% arose from FDPM instrumental error and the remainder was attributable to the complete-random-mixture model. The applicability of FDPM as an on-line sensor for powder-blending operations was further evaluated by analyzing grab samples taken directly from five locations of a 2-cu-ft Gallay blender at intervals of 5min within the blending process. FDPM results indicate that homogeneity was largely achieved in the first 10 min, during which the RSD of API content across five sampling locations decreased from 27% to 8%, and the RSD decreased to 5% after 25min of blending. Evolution of homogeneity within the blending process assessed through FDPM measurements was fit to the first-order model of particle blending further evidencing applicability for monitoring powder-blending processes. © 2004 Wiley-Liss, Inc. and the American Pharmacists Association

Determination of optical properties in semi-infinite turbid media using imaging measurements of frequency-domain photon migration obtained with an intensified charge-coupled device

Frequency-domain photon migration measurements across the surface of a tissue-mimicking, semi-infinite phantom are acquired via an intensified charge-coupled device (ICCD) detection system and used in conjunction with the diffusion approximation to determine the optical properties. The absorption and reduced scattering coefficients are determined least accurately when relative measurements of average light intensity I(rel)dc are employed either alone or in a combination with relative modulation amplitude data I(rel)ac and/or relative phase shift data theta(rel). The absorption and reduced scattering coefficients are found accurate to within 15 and 11%, respectively, of the values obtained from standard single-pixel measurements when theta(rel) measurements are employed alone or in combination with I(rel)ac data.

Investigation of structure factors in dense colloidal suspensions using frequency domain photon migration: polydisperse systems

Frequency domain photon migration (FDPM) technique was employed to investigate the structure factors of dense, polydisperse colloidal suspensions. The angle-integrated structure factors, 〈 S( q)〉, extracted from FDPM measurements of scattering properties at volume fractions ranging from 0.05 to 0.4, were compared with the values predicted from the polydisperse hard sphere Percus–Yevick (HSPY) model, as well as decoupling approximation (DA) and local monodisperse approximation (LMA) models that incorporated independently measured particle size information. Results show that the polydisperse HSPY model is the most suitable for accounting for particle interactions which predominately arise from volume exclusion effects. Furthermore, the influence of size polydispersity upon 〈 S( q)〉 is most significant at high volume fractions. The static structure factors at small wave vector q, S(0), were also assessed from dual wavelength FDPM measurements by using the small wave number approximation as well as the local monodisperse approximation. The measured S(0) agrees well with the values predicted by the polydisperse HSPY model.

Non-Invasive Hemodynamic Monitoring using near Infrared Frequency Domain Photon Migration in Rabbit Hemorrhagic Shock Model

Background Currently, there are no reliable, accurate, non-invasive methods of assessing hemodynamic parameters, tissue hemoglobin, and 0, content for physiologic monitoring. Recent advances in the field of optical diagnostics have lead to new imaging modalities capable of evaluating regional in-vivo tissue composition using its optical properties. The advantage of Frequency Domain Photon Migration (FDPM) over previously studied bulk NIR spectroscopy is that it allows for quantitative measurements of both light absorption and scattering in tissues. Devices based on FDPM principles have the potential to accurately and non-invasively monitor hemodynamics and other parameters in patients with shock and physiologic disturbances. Purpose This study investigated FDPM methods for monitoring shock and the response to treatment with volume expansion and pressors in a hemorrhagic rabbit model. The sensitivity of this monitoring method for discriminating changes in hemodynamic parameters compared to traditional invasive monitoring devises, as well as the ability to accurately measure hemoglobin and tissue 0, was assessed. Methods New Zealand White (NZW) rabbits (n=25) were used in this study. Aprototype FDPM device developed in our laboratory was used. Aprobe was placed on the inner thigh of the rabbits. The rabbits underwent concurrent traditional invasive monitoring including cardiac output (CO), pulmonary arterial pressure (PA), systemic blood pressure via arterial line, hemoglobin, and arterial and venous blood gases. Hemorrhage was performed via a syringe either in stages or in a single step. After measurements were obtained the animals were resuscitated with vasopressors, or with crystalloid solution. Results The FPDM measurements of tissue hemoglobin (Hb), Oxyhemoglobin (HbO,), water content and tissue 0, saturation were obtained. The tissue Hb (r=0.7) and oxygen content (r=0.6) closely correlated with the measurements made with invasive methods. The FDPM measurements also correlated with CO. Conclusion: Broad-bandwidth FDPM may be a reliable, accurate and non-invasive means of monitoring patients with hemorrhagic shock and their response to treatment. FDPM may potentially be adapted for use in early shock recognition for triage or in settings where aggressive invasive monitoring is problematic.

Sensitivity and depth penetration of continuous wave versus frequency-domain photon migration near-infrared fluorescence contrast-enhanced imaging.

The development of near-infrared fluorescent contrast agents and imaging techniques depends on the deep penetration of excitation light through several centimeters of tissue and the sensitive collection of the re-emitted fluorescence. In this contribution, the sensitivity and depth penetration of various fluorescence-enhanced imaging studies is surveyed and compared with current studies using continuous wave (CW) and frequency-domain photon migration (FDPM) measurements with planar wave illumination of modulated excitation light at 100 MHz and area collection of reemitted fluorescent light using a previously developed modulated intensified charge-coupled device camera system. Fluorescence was generated from nanomolar to micromolar solutions of indocyanine green (ICG) in a 100 microL volume submerged at 1-4 cm depths in a 1% Liposyn solution to mimic tissue scattering properties. Enhanced depth penetration and sensitivity are achieved with optimal filter rejection of excitation light, and FDPM rejection of background light is not achieved using CW methods. We show the ability to detect as few as 100 fmol of ICG from area illumination of 785 nm light (5.5 mW/cm2) and FDPM area collection of 830 nm fluorescent light generated from 3 cm below the phantom surface. The lowered noise floor of FDPM measurements enables greater sensitivity and penetration depth than comparable CW measurements.

Modification of Electrostatic Interaction by Rhodamine 6G Adsorption on Polystyrene Latex as Assessed by Frequency Domain Photon Migration

To demonstrate assessment of electrostatic interactions using first principles models, the isotropic scattering coefficients of negatively charged polystyrene latex were measured with frequency domain photon migration (FDPM) as the effective surface charge was changed by adsorbing positively charged surfactant, Rhodamine 6G (R6G). Measurements were conducted on dense suspensions (volume fraction = 0.186) at an ionic strength of 5 mM NaCl equivalent with varying amounts of positively charged R6G adsorbed on the particle surface. The FDPM measurements of isotropic scattering were then fitted to a model prediction obtained by solving the Ornstein−Zernike integral equation using the hard sphere Yukawa interaction model with mean spherical approximation as closure. The results show that the parameter estimate of surface charge decreased with adsorbed R6G showing the ability to use multiply scattered light to characterize electrostatic interaction in dense suspensions.

Three-dimensional, Bayesian image reconstruction from sparse and noisy data sets: near-infrared fluorescence tomography.

A method for inverting measurements made on the surfaces of tissues for recovery of interior optical property maps is demonstrated for sparse near-infrared (NIR) fluorescence measurement sets on large tissue-simulating volumes with highly variable signal-to-noise ratio. A Bayesian minimum-variance reconstruction algorithm compensates for the spatial variability in signal-to-noise ratio that must be expected to occur in actual NIR contrast-enhanced diagnostic medical imaging. Image reconstruction is demonstrated by using frequency-domain photon migration measurements on 256-cm(3) tissue-mimicking phantoms containing none, one, or two 1-cm(3) heterogeneities with 50- to 100-fold greater concentration of Indocyanine Green dye over background levels. The spatial parameter estimate of absorption owing to the dye was reconstructed from only 160 to 296 surface measurements of emission light at 830 nm in response to incident 785-nm excitation light modulated at 100 MHz. Measurement error of acquired fluence at fluorescent emission wavelengths is shown to be highly variable. Convergence and quality of image reconstructions are improved by Bayesian conditioning incorporating (i) experimentally determined measurement error variance, (ii) recursively updated estimates of parameter uncertainty, and (iii) dynamic zonation. The results demonstrate that, to employ NIR fluorescence-enhanced optical imaging for large volumes, reconstruction approaches must account for the large range of signal-to-noise ratio associated with the measurements.

Volume of pharmaceutical powders probed by frequency-domain photon migration measurements of multiply scattered light.

Using probability distribution analysis to describe the propagation of multiply scattered light between a point source and point detector located a known distance apart, mathematical expressions predicting the volume of sampled powder were determined for infinite and semi-infinite powder beds and then compared to experimental measurements of frequency-domain photon migration (FDPM). Our results show that the volume of powder sampled varies with optical properties and, when using FDPM techniques, with modulation frequency. For a typical measurement in lactose powder, the volume of powder sampled by multiply scattered light propagating between a 1000-microm-diameter point source and point detector pair separated by 8 mm is predicted to be 1.8 and 1.5 cm3 at modulated frequencies of 50 and 100 MHz, respectively. Experimental measurements in lactose powder confirm the predictions.

Three-dimensional fluorescence enhanced optical tomography using referenced frequency-domain photon migration measurements at emission and excitation wavelengths

The ultimate success of near-infrared optical tomography rests on the precise measurement of light propagation within tissues or random media, the accurate prediction of these measurements from a light propagation model, and an efficient three-dimensional solution of the inverse imaging problem. To date, optical tomography algorithms have focused on frequency-domain photon migration (FDPM) measurements of phase-delay and amplitude attenuation, which are reported relative to the incident light, even though phase-delay and amplitude of incident light are nearly impossible to measure directly. In this contribution, we examine referenced, fluorescence-enhanced frequency-domain photon migration measured at excitation and/or emission wavelengths and report on a measurement strategy to minimize measurement and calibration error for efficient coupling of data to a distorted Born iterative imaging algorithm. We examine three referencing approaches and develop associated inversion algorithms for (1) normalizing detected emission FDPM data to the predicted emission wave arising from a homogeneous medium, (2) referencing detected emission FDPM data to that detected at a reference point, and (3) referencing detected emission FDPM data to detected excitation FDPM data detected at a reference point. Our results show the latter approach to be practical while reducing the nonlinearity of the inverse problem. Finally, in light of our results, we demonstrate the method for eliminating the influence of source strength and instrument functions for effective fluorescence-enhanced optical tomography using FDPM.

Precise analysis of frequency domain photon migration measurement for characterization of concentrated colloidal suspensions

Frequency domain photon migration (FDPM) is a new method for characterization of concentrated colloidal suspensions using multiply scattered light. The ability of FDPM to determine size and interaction characteristics of the particulate phase of colloidal suspensions depends largely upon the accuracy and precision of FDPM-measured optical properties. In this work, FDPM measurements at multiple modulation frequencies and multiple source-to-detector distances are systematically analyzed for obtaining accurate and precise scattering properties of colloidal suspensions. Two different data analysis methods, multifrequency (MF) nonlinear regression and multidistance (MD) linear regression, and corresponding various strategies for fitting to the optical diffusion equation are investigated. The accuracy and precision of estimated scattering coefficients by different approaches are compared. Results show that MD linear regression with simultaneous regression of average intensity and phase shift or amplitude and phase shift data provides the best data analysis method since it provides not only accurate estimated parameters but also an accurate estimation of their uncertainties. Finally, an important criterion, derived from the photon diffusion model, is proposed for checking the consistency of measurement data and optimizing experimental conditions for FDPM characterization of multiply scattering materials.

Investigation of Particle Interactions in Dense Colloidal Suspensions Using Frequency Domain Photon Migration: Bidisperse Systems

The frequency domain photon migration (FDPM) technique was employed for investigation of particle interactions of dense, bidisperse, polystyrene samples of differring volume ratios of small to large particle sizes. The particle interactions of bidisperse systems were evaluated from the angular integrated structure factors, 〈S(q)〉 , which were extracted from FDPM measurements of scattering properties at high volume fractions ranging from 0.1 to 0.3. The FDPM-measured 〈S(q)〉 were then compared with theoretical predictions using the full binary hard sphere Percus−Yevick (HSPY) model, as well as the decoupling approximation (DA) and the local monodisperse approximation (LMA) models. The influence of differing volume ratios of colloid sizes on structure factors at differing wavelengths was also investigated. Results show that the interactions between small and large particles significantly impact the structure factors as well as the scattering properties in dense bidisperse colloidal suspensions. Furthermore, ...