Arjen Amelink

In vivo measurement of the local optical properties of tissue by use of differential path-length spectroscopy

We demonstrate the capability of differential path-length spectroscopy (DPS) to determine the local optical properties of tissue in vivo. DPS measurements on bronchial mucosa are analyzed and yield information on the local blood oxygenation, blood content, average microvessel diameter, and wavelength dependence of the reduced scattering coefficient. Our data collected to date show that cancerous bronchial mucosa has a lower capillary oxygenation and a larger average capillary diameter than normal bronchial mucosa.

Measurement of the local optical properties of turbid media by differential path-length spectroscopy

We report on the development of an optical-fiber-based diagnostic tool with which to determine the local optical properties of a turbid medium. By using a single fiber in contact with the medium to deliver and detect white light, we have optimized the probability of detection of photons scattered from small depths. The contribution of scattered light from greater depths to the signal is measured and subtracted with an additional fiber, i.e., a collection fiber, to yield a differential backscatter signal. Phantoms demonstrate that, when photons have large mean free paths compared with the fiber diameter, single scattering dominates the differential backscatter signal. When photons have small mean free paths compared with the fiber diameter, the apparent path length of the photons that contribute to the differential backscatter signal becomes approximately equal to 4/5 of the fiber diameter. This effect is nearly independent of the optical properties of the sample under investigation.

Optical biopsy of breast tissue using differential path-length spectroscopy

Differential path-length spectroscopy (DPS) was used to determine the local optical properties of breast tissue in vivo. DPS measurements were made on healthy and malignant breast tissue using a fibre-optic needle probe, and were correlated to the histological outcome of core-needle biopsies taken from the same location as the measurements. DPS yields information on the local tissue blood content, the local blood oxygenation, the average micro-vessel diameter, the beta-carotene concentration and the scatter slope. Our data show that malignant breast tissue is characterized by a significant decrease in tissue oxygenation and a higher blood content compared to normal breast tissue.

Single-scattering spectroscopy for the endoscopic analysis of particle size in superficial layers of turbid media

We report on the development of an optical-fiber-based diagnostic tool that is sensitive to single-scattering events close to the fiber-optic probe tip. By using a single fiber to deliver and detect white light we optimised the detection probability of singly scattered photons from small depths. The sampling depth of this delivery-and-collection fiber was investigated by use of a tissue phantom. We found that for our phantom 90% of the single-scattering signal in the delivery-and-collection fiber originated from less than 200 microm from the fiber tip. The contribution of multiply scattered light from a greater depth to the signal was measured with an additional collection fiber. Several tissue phantoms demonstrated our fiber-optic probes sensitivity to light scattering from superficial layers of tissue and thereby its potential to detect superficial precancerous epithelial lesions.

Monte Carlo analysis of single fiber reflectance spectroscopy: photon path length and sampling depth

Single fiber reflectance spectroscopy is a method to noninvasively quantitate tissue absorption and scattering properties. This study utilizes a Monte Carlo (MC) model to investigate the effect that optical properties have on the propagation of photons that are collected during the single fiber reflectance measurement. MC model estimates of the single fiber photon path length (L(SF)) show excellent agreement with experimental measurements and predictions of a mathematical model over a wide range of optical properties and fiber diameters. Simulation results show that L(SF) is unaffected by changes in anisotropy (g epsilon [0.8, 0.9, 0.95]), but is sensitive to changes in phase function (Henyey-Greenstein versus modified Henyey-Greenstein). A 20% decrease in L(SF) was observed for the modified Henyey-Greenstein compared with the Henyey-Greenstein phase function; an effect that is independent of optical properties and fiber diameter and is approximated with a simple linear offset. The MC model also returns depth-resolved absorption profiles that are used to estimate the mean sampling depth (Z(SF)) of the single fiber reflectance measurement. Simulated data are used to define a novel mathematical expression for Z(SF) that is expressed in terms of optical properties, fiber diameter and L(SF). The model of sampling depth indicates that the single fiber reflectance measurement is dominated by shallow scattering events, even for large fibers; a result that suggests that the utility of single fiber reflectance measurements of tissue in vivo will be in the quantification of the optical properties of superficial tissues.

Empirical model of the photon path length for a single fiber reflectance spectroscopy device.

A reflectance spectroscopic device that utilizes a single fiber for both light delivery and collection has advantages over classical multi-fiber probes. This study presents a novel empirical relationship between the single fiber path length and the combined effect of both the absorption coefficient, mua (range: 0.1-6 mm-1), and the reduced scattering coefficient, micros (range: 0.3 - 10 mm-1), for different anisotropy values (0.75 and 0.92), and is applicable to probes containing a wide range of fiber diameters (range: 200-2000 microm). The results indicate that the model is capable of accurately predicting the single fiber path length over a wide range (r = 0.995; range: 180-3940 microm) and predictions do not show bias as a function of either microa or micros .

Integration of single-fiber reflectance spectroscopy into ultrasound-guided endoscopic lung cancer staging of mediastinal lymph nodes

We describe the incorporation of a single-fiber reflectance spectroscopy probe into the endoscopic ultrasound fine-needle aspiration (EUS-FNA) procedure utilized for lung cancer staging. A mathematical model is developed to extract information about the physiological and morphological properties of lymph tissue from single-fiber reflectance spectra, e.g., microvascular saturation, blood volume fraction, bilirubin concentration, average vessel diameter, and Mie slope. Model analysis of data from a clinical pilot study shows that the single-fiber reflectance measurement is capable of detecting differences in the physiology between normal and metastatic lymph nodes. Moreover, the clinical data show that probe manipulation within the lymph node can perturb the in vivo environment, a concern that must be carefully considered when developing a sampling strategy. The data show the feasibility of this novel technique; however, the potential clinical utility has yet to be determined.

Measurement of the reduced scattering coefficient of turbid media using single fiber reflectance spectroscopy: Fiber diameter and phase function dependence

This paper presents a relationship between the intensity collected by a single fiber reflectance device (RSF) and the fiber diameter (dfib) and the reduced scattering coefficient ( μs′) and phase function (p(θ)) of a turbid medium. Monte Carlo simulations are used to identify and model a relationship between RSF and dimensionless scattering ( μs′dfib). For μs′dfib > 10 we find that RSF is insensitive to p(θ). A solid optical phantom is constructed with μs′ ≈ 220 mm−1 and is used to convert RSF of any turbid medium to an absolute scale. This calibrated technique provides accurate estimates of μs′ over a wide range ([0.05 – 8] mm−1) for a range of dfib ([0.2 – 1] mm).

Monitoring ALA-induced PpIX Photodynamic Therapy in the Rat Esophagus Using Fluorescence and Reflectance Spectroscopy

The presence of phased protoporphyrin IX (PpIX) bleach kinetics has been shown to correlate with esophageal response to 5‐aminolevulinic acid‐based photodynamic therapy (ALA‐PDT) in animal models. Here we confirm the existence of phased PpIX photobleaching by increasing the temporal resolution of the fluorescence measurements using the therapeutic illumination and long wavelength fluorescence detection. Furthermore fluorescence differential pathlength spectroscopy (FDPS) was incorporated to provide information on the effects of PpIX and tissue oxygenation distribution on the PpIX bleach kinetics during illumination. ALA at a dose of 200 mg kg−1 was orally administered to 15 rats, five rats served as control animals. PDT was performed at an in situ measured fluence rate of 75 mW cm−2 using a total fluence of 54 J cm−2. Forty‐eight hours after PDT the esophagus was excised and histologically examined for PDT‐induced damage. Fluence rate and PpIX photobleaching at 705 nm were monitored during therapeutic illumination with the same isotropic probe. A new method, FDPS, was used for superficial measurement on saturation, blood volume, scattering characteristics and PpIX fluorescence. Results showed two‐phased PpIX photobleaching that was not related to a (systematic) change in esophageal oxygenation but was associated with an increase in average blood volume. PpIX fluorescence photobleaching measured using FDPS, in which fluorescence signals are only acquired from the superficial layers of the esophagus, showed lower rates of photobleaching and no distinct phases. No clear correlation between two‐phased photobleaching and histologic tissue response was found. This study demonstrates the feasibility of measuring fluence rate, PpIX fluorescence and FDPS during PDT in the esophagus. We conclude that the spatial distribution of PpIX significantly influences the kinetics of photobleaching and that there is a complex interrelationship between the distribution of PpIX and the supply of oxygen to the illuminated tissue volume.

Confidence intervals on fit parameters derived from optical reflectance spectroscopy measurements

We validate a simple method for determining the confidence intervals on fitted parameters derived from modeling optical reflectance spectroscopy measurements using synthetic datasets. The method estimates the parameter confidence intervals as the square roots of the diagonal elements of the covariance matrix, obtained by multiplying the inverse of the second derivative matrix of chi2 with respect to its free parameters by chi2/v, with v the number of degrees of freedom. We show that this method yields correct confidence intervals as long as the model used to describe the data is correct. Imperfections in the fitting model introduces a bias in the fitted parameters that greatly exceeds the estimated confidence intervals. We investigate the use of various methods to identify and subsequently minimize the bias in the fitted parameters associated with incorrect modeling.