In vivo determination of optical properties of normal and tumor tissue with white light reflectance and an empirical light transport model during endoscopy.

Determination of tissue optical properties is fundamental for application of light in either therapeutical or diagnostics procedures. In the present work we implemented a spatially resolved steady-state diffuse reflectance method where only two fibers (one source and one detector) spaced 2.5 mm apart are used for the determination of the optical properties. The method relies on the spectral characteristics of the tissue chromophores (water, dry tissue, and blood) and the assumption of a simple wavelength dependent expression for the determination of the reduced scattering coefficient. Because of the probe dimensions the method is suited for endoscopic measurements. The method was validated against more traditional models, such as the diffusion theory combined with adding doubling for in vitro measurements of bovine muscle. Mean and standard deviation of the absorption coefficient and the reduced scattering coefficient at 630 nm for normal mucosa were 0.87+/-0.22 cm(-1) and 7.8+/-2.3 cm(-1), respectively. Cancerous mucosa had values 1.87+/-1.10 cm(-1) and 8.4+/-2.3 cm(-1), respectively. These values are similar to data presented by other authors. Blood perfusion was the main variable accounting for differences in the absorption coefficient between the studied tissues.

Pulsed photothermal radiometry in optically transparent media containing discrete optical absorbers.

A description of heat transport by conduction and radiation in inhomogeneous materials following absorption of a brief optical pulse is presented, and investigated experimentally using pulsed photothermal radiometry (PPTR). The model indicates that the role of radiation as an intramedium heat transfer modality increases with increasing temperatures and decreasing infrared (IR) absorption of the medium. However, for the range of conditions analysed in this study, conductive transfer dominates. Thus, the inclusion of radiation does not significantly perturb the internal temperature profiles, although it does influence the radiometric emission from the sample, and hence the PPTR signal. The thermal confinement effects described in this study may be relevant in photomedicine, for example in pulsed laser irradiation of tissues containing small absorbing targets.

Determination of optical properties of turbid media using pulsed photothermal radiometry.

Pulsed photothermal radiometry (PPTR) measures blackbody radiation emitted by a sample after absorption of an optical pulse. Three techniques for obtaining the absorption coefficient of absorbing-only, semi-infinite samples are examined and shown to give comparable results. An analytic theory for the time dependence of the PPTR signal in semi-infinite scattering and absorbing media has been derived and tested in a series of controlled gel phantoms. This theory, based on the diffusion approximation of the radiative transport equation, is shown to model the time course of the detected signal accurately. Furthermore, when the incident fluence is known, the theory can be used in a non-linear, two-parameter fitting algorithm to determine the absorption and reduced scattering coefficients of a turbid sample with an accuracy of 10-15% for transport albedos ranging from 0.42-0.88.

Modeling optical and thermal distributions in tissue during laser irradiation.

The propagation of light energy in tissues is an important problem in phototherapy, especially with the increased use of lasers as light sources. Often a slight difference in delivered energy separates a useless, efficacious, or disastrous treatment. Methods are presented for experimental characterization of the optical properties of a tissue and computational prediction of the distribution of light energy within a tissue. A standard integrating sphere spectrophotometer measured the total transmission, Tt, total reflectance, Rt, and the on-axis transmission, Ta, for incident collimated light that propagated through the dermis of albino mouse skin, over the visible spectrum. The diffusion approximation solution to the one-dimensional (1-D) optical transport equation computed the expected Tt and Rt for different combinations of absorbance, k, scattering, s, and anisotropy, g, and by iterative comparison of the measured and computed Tt and Rt values converged to the intrinsic tissue parameters. For example, mouse dermis presented optical parameters of 2.8 cm-1, 239 cm-1, and 0.74 for k, s, and g, respectively, at 488 nm wavelength. These values were used in the model to simulate the optical propagation of the 488-nm line of an argon laser through mouse skin in vivo. A 1-D Green's function thermal diffusion model computed the temperature distribution within the tissue at different times during laser irradiation. In vitro experiments showed that the threshold temperature range for coagulation was 60 degrees-70 degrees C, and the kinetics were first order, with a temperature-dependent rate constant that obeyed an Arrhenius relation (molar entropy 276 cal/mol-degrees K, molar enthalpy 102 kcal/mol). The model simulation agreed with the corresponding in vivo experiment that a 2-s pulse at 55 W/cm2 irradiance will achieve coagulation of the skin.

Laser thrombolysis using long pulse frequency-doubled Nd:YAG lasers.

BACKGROUND AND OBJECTIVE: Laser thrombolysis is a means for clearing blood clots in occluded arteries. Many researchers have studied the mechanisms of clot ablation, and research clinicians have used the technique to treat myocardial infarction with a number of different laser systems. Specifically, a 1-microsec pulsed dye laser has been used clinically to remove blood clots in coronary arteries. As a comparative study, the ablation characteristics of lasers with pulse durations in the ranges of 50-150 microsec and 2-10 msec were investigated. Two frequency-doubled Nd:YAG lasers at 532 nm were used in this study. Ablation threshold and ablation efficiency of gel phantoms and thrombus using these two lasers were measured and compared with the results of the pulsed dye laser. The pulsed dye laser in this study operated at 522 nm. STUDY DESIGN/MATERIALS AND METHODS: Gelatin samples with 150 cm(-1) absorption coefficient at 532 nm and animal clot were confined to 3-mm silicone tubes to measure ablation parameters. Additional samples with 150 cm(-1) absorption coefficient at 522 nm were prepared for use with the pulsed dye laser. A fluorescence technique and photographic bubble detection were used to determine ablation threshold. A spectrophotometric technique was used to determine ablation efficiency. RESULTS: The ablation threshold of the gel phantoms for all three lasers was determined to be 17 +/- 2 mJ/mm(2). Ablation efficiency for the gel phantoms was 1.7 +/- 0.1 microg/mJ. Clot had an ablation efficiency of 2.9 +/- 1.0 microg/mJ. CONCLUSIONS: Ablation threshold and efficiency are independent of laser pulse duration for 1-microsec, 50-150-microsec, and 2-10-msec pulses (P < 0.05).

Pressure impulses during microsecond laser ablation.

The collapse of laser-induced cavitation bubbles creates acoustic transients within the surrounding medium and also pressure impulses to the ablation target and light-delivery fiber during microsecond laser ablation. The impulses are investigated here with time-resolved flash photography, and they are found to occur whether or not the light-delivery fiber is in contact with the target. We demonstrate that the impulses depend primarily on the energy stored in the cavitation bubble. They are not directly dependent on the mode of light delivery (contact versus noncontact), and they are also not directly correlated to the other acoustic transients. The pressure impulses do seem to be associated with the bubble-driven jet formation caused by the bubble collapse.

Accuracies of the diffusion approximation and its similarity relations for laser irradiated biological media.

The accuracy of the diffusion approximation is compared with more accurate solutions for describing light interaction with biological tissues. Generally the diffusion approximation underestimates the light distribution in the surface region, and, for high albedos, it significantly underestimates the fluence rate. This difference is only a few percent for albedos of less than 0.5 due to the dominance of collimated light. As the anisotropy of scattering increases, deviations increase. In general, fluxes can be computed more accurately with the diffusion approximation than fluence rates. For anisotropic scattering, better results can be obtained by simple transforms of optical coefficients using the similarity relations. The similarity relations improve flux calculations, but computed fluence rates have substantial errors for high albedo and the large index of refraction differences at the surface.

Determining the optical properties of turbid mediaby using the adding-doubling method.

A method is described for finding the optical properties (scattering, absorption, and scattering anisotropy) of a slab of turbid material by using total reflection, unscattered transmission, and total transmission measurements. This method is applicable to homogeneous turbid slabs with any optical thickness,albedo, or phase function. The slab may have a different index of refraction from its surroundings and may or may not be bounded by glass. The optical properties are obtained by iterating an adding-doubling solution of the radiative transport equation until the calculated values of the reflection and transmission match the measured ones. Exhaustive numerical tests show that the intrinsic error in the method is < 3% when four quadrature points are used.

Light distributions in artery tissue: Monte Carlo simulations for finite-diameter laser beams.

Finite-width light distributions in arterial tissue during Argon laser irradiation (476 nm) are simulated using the Monte Carlo method. Edge effects caused by radial diffusion of the light extend +/- 1.5 mm inward from the perimeter of a uniform incident beam. For beam diameters exceeding 3 mm the light distribution along the central axis can be described by the one-dimensional solution for an infinitely wide beam. The overlapping edge effects for beam diameters smaller than 3 mm reduce the penetration of the irradiance in the tissue. The beam profile influences the light distribution significantly. The fluence rates near the surface for a Gaussian beam are two times higher on the central axis and decrease faster radially than for a flat profile. The diverging light from a fiber penetrates tissue in a manner similar to collimated light.

Threshold and ablation efficiency studies of microsecond ablation of gelatin under water.

BACKGROUND AND OBJECTIVE: Laser thrombolysis is the selective ablation of thrombus occluding vessels by microsecond pulsed laser irradiation. To achieve efficient ablation of thrombus, the optimal wavelength, spot size, and pulse energy need to be determined. STUDY DESIGN/MATERIALS AND METHODS: A gelatin-based thrombus model confined in 3 mm inner diameter tubes was ablated under water using a 1 microsecond pulsed dye laser. Wavelength studies were conducted by varying the absorption of the gelatin between 10-2000 cm-1 corresponding to the waveband between 400-600 nm on the absorption spectrum of thrombus. A unique spectrophotometric method was developed to measure the ablated mass. An acoustic method was used to measure ablation thresholds under water as a function of absorption. RESULTS: The mass removed per pulse per unit energy was nearly equal over an absorption range of 100-1000 cm-1 at pulse energies above threshold. Mass removal increased linearly with pulse energy but did not have a direct relationship with radiant exposure. Ablation thresholds indicate that the gelatin needed to be heated only to 100 degrees C for ablation to commence. Ablation masses measured were an order of magnitude higher than those predicted by thermal ablation models. CONCLUSION: The results suggest that any wavelength between 410-590 nm can be used for effective thrombolysis. The ablation efficiency depends on the total energy delivered rather than the radiant exposure. The high ablation efficiencies suggest a dominance of the mechanical action of vapor bubbles over thermal ablation in the ablation process.

Drug delivery with microsecond laser pulses into gelatin.

Photo acoustic drug delivery is a technique for localized drug delivery by laser-induced hydrodynamic pressure following cavitation bubble expansion and collapse. Photoacoustic drug delivery was investigated on gelatin-based thrombus models with planar and cylindrical geometries by use of one microsecond laser pulses. Solutions of a hydrophobic dye in mineral oil permitted monitoring of delivered colored oil into clear gelatin-based thrombus models. Cavitation bubble development and photoacoustic drug delivery were visualized with flash photography. This study demonstrated that cavitation is the governing mechanism for photoacoustic drug delivery, and the deepest penetration of colored oil in gels followed the bubble collapse. Spatial distribution measurements revealed that colored oil could be driven a few millimeters into the gels in both axial and radial directions, and the penetration was less than 500 µm when the gelatin structure was not fractured.

Enhanced laser thrombolysis with photomechanical drug delivery: an in vitro study.

BACKGROUND AND OBJECTIVE: Current techniques for laser thrombolysis are limited because they can not completely clear thrombotic occlusions in arteries, typically leaving residual thrombus on the walls of the artery. The objective of this study was to investigate the possibility of using photomechanical drug delivery to enhance laser thrombolysis by delivering drugs into mural thrombus during laser thrombolysis. STUDY DESIGN/MATERIALS AND METHODS: Three experimental protocols were performed in vitro to quantitatively compare the effectiveness of thrombolysis by 1) constant infusion of drug, 2) laser thrombolysis, and 3) photomechanical drug delivery. A fiber-optic flushing catheter delivered drug (a solution of 1 microm fluorescent microspheres) and light ( a 1 micros pulsed dye laser) into a gelatin-based thrombus model. The process of laser-thrombus interaction was visualized using flash photography and the laser-induced pressure waves were measured using an acoustic transducer. RESULTS: Lumen sizes generated by mechanically manipulating the catheter through the thrombus were smaller than those generated by laser ablation. The microspheres could be driven several hundred microns into the mural thrombus. CONCLUSION: Photomechanical drug delivery has potential for enhancement of laser thrombolysis. Two mechanisms seem to be involved in photomechanical drug delivery: 1) mural deposition of the drug at the ablation site and 2) increased exposure of the thrombus surface area to the drug.

Mechanical properties of coagulated albumin and failure mechanisms of liver repaired with the use of an argon-beam coagulator with albumin.

Hemostasis in the traumatized liver has been achieved by thermally denaturing topically applied albumin. In this article, the mechanical properties of liver and denatured albumin (solder) were measured, and the failure methods of liver repaired with albumin were identified. The ultimate tensile strength and Young's modulus were measured for healthy liver (N = 20) and thermally damaged liver (N = 20). The ultimate tensile strength and Young's modulus were measured for three concentrations of coagulated albumin (25, 38, and 53%) in a single layer and for two layers of denatured 38% albumin. Failure under tension of argon-beam coagulator soldered liver on the parenchymal surface (N = 30) with 38% albumin in two layers had a 70% occurrence for tearing at a mean stress of 39 kPa and a 23% occurrence for shearing at a mean stress of 7 kPa. Liver repaired on the interior surface (N = 11) failed in tension by tearing (64%) at a mean stress of 34 kPa and by shearing (36%) at a mean stress of 6 kPa. Argon-beam coagulator soldering with 38% albumin took 6 s/cm(2) for two layers of solder and gave the best balance of usability, strength, and matching of mechanical properties with those of the liver.

Iterative reconstruction algorithm for optoacoustic imaging.

Optoacoustic imaging is based on the generation of thermoelastic stress waves by heating an object in an optically heterogeneous medium with a short laser pulse. The stress waves contain information about the distribution of structures with preferential optical absorption. Detection of the waves with an array of broadband ultrasound detectors at the surface of the medium and applying a backprojection algorithm is used to create a map of absorbed energy inside the medium. With conventional reconstruction methods a large number of detector elements and filtering of the signals are necessary to reduce backprojection artifacts. As an alternative this study proposes an iterative procedure. The algorithm is designed to minimize the error between measured signals and signals calculated from the reconstructed image. In experiments using broadband optical ultrasound detectors and in simulations the algorithm was used to obtain three-dimensional images of multiple optoacoustic sources. With signals from a planar array of 3x3 detector elements a significant improvement was observed after about 10 iterations compared to the simple radial backprojection. Compared to conventional methods using filtered backprojection, the iterative method is computationally more intensive but requires less time and instrumentation for signal acquisition.

Light scattering in Intralipid-10% in the wavelength range of 400-1100 nm.

The absorption, scattering, and anisotropy coefficients of the fat emulsion Intralipid-10% have been measured at 457.9, 514.5, 632.8, and 1064 nm. The size and shape distributions of the scattering particles in Intralipid-10% were determined by transmission electron microscopy. Mie theory calculations performed by using the particle size distribution yielded values for the scattering and anisotropy coefficients from 400 to 1100 nm. The agreement with experimental values is better than 6%.

A self-heated thermistor technique to measure effective thermal properties from the tissue surface.

A microcomputer based instrument to measure effective thermal conductivity and diffusivity at the surface of a tissue has been developed. Self-heated spherical thermistors, partially embedded in an insulator, are used to simultaneously heat tissue and measure the resulting temperature rise. The temperature increase of the thermistor for a given applied power is a function of the combined thermal properties of the insulator, the thermistor, and the tissue. Once the probe is calibrated, the instrument accurately measures the thermal properties of tissue. Conductivity measurements are accurate to 2 percent and diffusivity measurements are accurate to 4 percent. A simplified bioheat equation is used which assumes the effective tissue thermal conductivity is a linear function of perfusion. Since tissue blood flow strongly affects heat transfer, the surface thermistor probe is quite sensitive to perfusion.

Measurements and calculations of the energy fluence rate in a scattering and absorbing phantom at 633 nm.

We have studied the influence of absorption, scattering, and refractive index of a phantom medium in conjunction with various beam diameters on the penetration depth of light at 633 nm. We used mixtures of Intralipid 10% (scattering medium) and Evans blue (absorbing medium). Measurements were performed in media with a scattering coefficient of 1 mm(-1), an anisotropy factor of 0.71, absorption coefficients of 1.3 x 10(-3), 0.01, and 0.05 mm(-1), and a refractive index of 1.33. The experimental results were compared with an analytical solution of the fluence rate based on diffusion theory. We found good agreement (deviations of <10%) between theory and experiment for incident beam diameters between 10 and 60 mm.

Double-integrating-sphere system for measuring the optical properties of tissue.

A system is described and evaluated for the simultaneous measurement of the intrinsic optical properties of tissue: the scattering coefficient, the absorption coefficient, and the anisotropy factor. This system synthesizes the theory of two integrating spheres and an intervening scattering sample with the inverse adding-doubling algorithm, which employs the adding-doubling solution of the radiative transfer equation to determine the optical properties from the measurement of the light flux within each sphere and of the unscattered transmission. The optical properties may be determined simultaneously, which allows for measurements to be made while the sample undergoes heating, chemical change, or some otherexternal stimulus. An experimental validation of the system with tissue phantoms resulted in the determination of the optical properties with a < 5% deviation when the optical density was between 1 and 10 and the albedo was between 0.4 and 0.95.