Optical Properties of Tissues after Laser Treatments in the Wavelength Range of nm

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1 Optical Properties of Tissues after Laser Treatments in the Wavelength Range of nm Katsunori Ishii, Akinori Kimura, Kunio Awazu Sustainable Energy and Environmental Engineering, Graduate School of Engineering, Osaka University (2-1-A14 Yamada-Oka, Suita, Osaka, Japan, ) ABSTRACT In laser medicine, the accurate knowledge about the optical properties of target tissue is important for the understanding prediction of propagation and distribution of light in tissues. Light propagations, absorption and scattering, changes by the kinetic changes of optical properties in laser irradiations. This problem is clinically very important for the realization of safe laser treatments because the understandings of optical properties by several laser parameters can realize the preestimated treatment effects. The objective of this study is determination of the optical properties of treated tissues for the safe laser treatments. In this paper, we examined to determine the optical properties changes of a coagulated tissue after the Er:YAG laser irradiations by using double integrating sphere system and inverse adding-doubling method in the wavelength range of nm. After laser irradiations with a variety of irradiation parameters, the absorption coefficient of a treated tissue decreased and the reduced scattering coefficient of a treated tissue increase. In carbonization, observed in 30 sec-300 mj irradiation, the absorption coefficient of a treated tissue re-increased and the reduced scattering coefficient of a treated tissue re-decrease. The changes of optical properties should be accounted for while planning the therapeutic procedure for the realization of safe laser treatments. Keywords: optical property, absorption coefficient, scattering coefficient, double integrating sphere, laser treatment, inverse adding-doubling (IAD), Er:YAG laser 1. INTRODUCTION In laser medicine, the accurate knowledge about the optical properties (scattering coefficient; s, absorption coefficient; a, scattering anisotropy; g) of target tissue is important for the understanding prediction of propagation and distribution of light in tissues. This information is valuable for the optimization of a variety of diagnostic and therapeutic biomedical applications of light. For example, photodynamic therapy employs light to activate a phototoxin that produces tumor destruction. The optimal clinical implementation of this technique depends critically on the distribution of light within the irradiated tissue. Similarly, laser ablation of tissue and laser-induced tissue damage are strongly dependent on the optical properties of the target. Thus it is likely that the optical properties of tissues at near-ultraviolet, visible, nearinfrared (NIR) and mid-infrared (MIR) wavelengths will become increasingly important in the future as more biomedical applications of lasers are developed. Recently the optical properties of various normal and pathologic tissues have been determined at single wavelength or over broad wavelength range. However, to our knowledge, there are no experimental studies about alternation of the tissues considered from the aspect of optical properties after many treatments (laser coagulation therapy, photo dynamic therapy, regenerative medicine, etc.) Double integrating sphere system [1-5] with an intervening sample is designed for non-invasive measurement of the optical properties of biological tissues. Using integrating sphere as both a diffuse illumination source and a detector provides a technically simple measurement apparatus. Tissue samples are sandwiched between two glass plates to minimize the usual irregularities in the tissue surface, and approximate the Fresnel reflection. This measurement system can determine the diffuse reflectance, diffuse and collimated transmittances of the samples. From these experimental data, the set of optical properties can be calculated by some programs for optical properties determination. Some studies employ indirect methods to determine optical properties, such as Kubelka-Munk, are limited their accuracy and ability to predict soft tissue behavior. On the other hand, Inverse Monte Carlo methods [3-8] can be used to determine the optical properties separately over broad spectral range, without limitation of their accuracy. It has the ability to compensate the conditions of measurements, sample geometries and sources of experimental error. However, large computing time needed to acquire adequate signal to noise ratio from the Monte Carlo simulations and ensure Biophotonics: Photonic Solutions for Better Health Care, edited by Jürgen Popp, Wolfgang Drexler, Valery V. Tuchin, Dennis L. Matthews, Proc. of SPIE Vol. 6991, 69912F, (2008) /08/$18 doi: / SPIE Digital Library -- Subscriber Archive Copy Proc. of SPIE Vol F-1

2 proper convergence to the correct optical properties. The inverse adding-doubling (IAD) method [9-11] developed by Dr. Scott Prahl et al, one of the most famous programs of calculating optical properties, is sufficiently fast for iterated solutions on current computing and flexible that anisotropic scattering and internal reflection at the boundaries may be included. This combined measurement system is able to determine the set of optical properties with high accuracy at short time. The changes of optical properties by laser treatments are particularly interesting [12,13]. Light propagations, absorption and scattering, changes by the kinetic changes of optical properties in laser irradiations. This problem is clinically very important for the realization of safe laser treatments because the understandings of optical properties by several laser parameters can realize the pre-estimated treatment effects. The objective of this study is determination of the optical properties of treated tissues for the safe laser treatments. In this paper, we examined to determine the optical properties changes of a coagulated tissue after the Er:YAG laser irradiations by using double integrating sphere system and IAD method in the wavelength range of nm. 2. MATERIALS AND METHODS 2.1 Sample Preparation First, we measured the optical properties of a standard sample, Intralipid, to check the validity of the system used in this study. An aqueous suspension of 20% Intralipid (FB-01IL20, Terumo, Japan) was diluted by 10% to 1.25% with water. These solutions were sealed between two slide glasses with the optical pass of 1 mm. Then, we measured the optical properties of a laser treated tissue. Chicken breast tissue was cut into the thickness of 5 mm using a surgical knife. Cut tissues were placed on a slide glass. After laser irradiations mentioned below, the laser treated tissue was covered with another slide glass to minimize the usual irregularities in the tissue surface. 2.2 Laser Irradiation We used a Er:YAG laser with a wavelength of 2.94 m, a repetition rate of 10 pps (Erwin Adverl, Morita, Japan) [14]. The laser beam was guided through the optical hollow fiber and irradiated without focusing. The irradiation area was 38.5 mm 2. The energy per pulse was within mj and the irradiation time was within 0-30 sec. 2.3 Integrating Sphere Measurements Double integrating sphere system with an intervening sample is designed for non-invasive measurement of the optical properties of biological tissues. This is also a convenient tool since it measure the diffuse reflectance (Rd) and total transmittance (Tt) simultaneously. Using integrating sphere as both a diffuse illumination source and a detector provides a technically simple measurement apparatus. A schematic of the experimental setup was presented in Figure 1. We employed balanced deuterium tungsten halogen source ( nm wavelength range, DH-2000-BAL, Ocean optics) combined with high-powered halogen light Inverse Adding Doubling Calculator Optical Properties ( a, s ) Reflectance Transmittance PC High resolution spectrometer Light source Deuterium + Halogen Sample Fig. 1. Experimental setup of double integrating sphere system Proc. of SPIE Vol F-2

3 source ( nm wavelength range, HL-2000-HP, Ocean Optics). Specimens were placed between two 38.1 mm diameter integrating spheres (FOIS-1, Ocean Optics) which were coated with diffusely reflective material, Spectralon TM. The entrance port size of reflectance sphere was 8 mm in diameter, and sample port of both spheres was 9.5 mm in diameter. The beam-illuminated area was 3 mm in diameter on the sample with glass slides. The incident light was diffusely reflected from sample surface and diffusely or collimated transmit sample. Then the light was multiply scattered in the spheres and recorded by high-resolution spectrometer (HR-4000, Ocean Optics) as Rd and Tt, respectively. From these experimental data, the set of optical properties were calculated by IAD method. 2.4 Inverse Adding-Doubling Method We employed IAD technique to calculate optical properties of samples from measured value of Rd and Tt. IAD is a technique developed by Scott Prahl et al, which uses adding-doubling method to figure out the optical properties of slabs of material from the observed transmission and reflection. This program is sufficiently fast for iterated solutions on current computing and flexible that anisotropic scattering and internal reflection at the boundaries may be included. This combined measurement system is able to determine the set of optical properties with high accuracy at short time. Additionally, up-to-date program was added a Monte Carlo calculation so that light lost out the edges of a sample can be estimated. This function allows to determine optical properties more accurately [9,10]. Fig. 2. The optical properties of the Intralipid solution with the concentration from 1.25 % to 10%. (a) absorption coefficient spectra, (b) reduced scattering spectra Proc. of SPIE Vol F-3

4 3. RESULTS AND DISCUSSION Figure 2(a) showed the absorption coefficient spectra of the Intralipid solution with the concentration from 1.25 % to 10%. Absorption coefficient spectra did not show significant differences in the spectral range examined. Absorption coefficients slightly decreased with increasing the concentrations of Intralipid solution. Figure 2(b) showed the reduced scattering coefficient spectra of the Intralipid solution with the concentration from 1.25 % to 10%. Reduced scattering coefficient spectra generally decreased with increasing wavelength, and reduced scattering coefficients gradually increased with increasing the concentrations of Intralipid solution, consistent with Rayleigh scattering theory and previously published results. Figure 3(a) showed the absorption coefficient spectra of the chicken breast tissues after Er:YAG laser coagulations. Irradiation time was 30 sec. Irradiated energies were 100, 200 and 300 mj. After laser irradiation with 100 mj and 200 mj, the absorption coefficient spectra from 350 nm to 1000 nm slightly decreased. After laser irradiation with 300 mj, the absorption coefficient spectrum from 650 nm to 1000 nm slightly decreased, however the absorption coefficient spectrum from 350 nm to 650 nm gradually increased with decreasing wavelength. Especially the absorption coefficient spectrum from 360 nm to 510 nm increase more than that of non-irradiation. Fig. 3. The optical properties of the chicken breast tissues after Er:YAG laser coagulations. Irradiation time is 30 sec. Irradiated energies are 100, 200 and 300 mj. (a) absorption coefficient spectra, (b) reduced scattering spectra Proc. of SPIE Vol F-4

5 Figure 3(b) showed the reduced scattering coefficient spectra of the chicken breast tissues after Er:YAG laser coagulations. Irradiation time was 30 sec. Irradiated energies were 100, 200 and 300 mj. After laser irradiation with 100 mj and 200 mj, reduced scattering coefficient spectra from 350 nm to 1000 nm gradually increased with increasing irradiation energy. The highest reduced scattering coefficient in 100 mj irradiation was found at 440 nm and the highest reduced scattering coefficient in 200 mj irradiation was found at 450 nm. After laser irradiation with 300 mj, reduced scattering coefficient spectra from 450 nm to 1000 nm increased and the reduced scattering coefficient values were lower than that of the laser irradiation with 200 mj. The highest reduced scattering coefficient in 300 mj irradiation was found at 470 nm. After laser irradiation with 300 mj, carbonization was observed on the surface of coagulated tissue. Reincreasing of absorption coefficient spectrum and re-reducing of reduced scattering coefficient spectrum observed in 300 mj irradiation were estimated by the influence on carbonization. Figure 4(a) showed the absorption coefficient spectra of the chicken breast tissues after Er:YAG laser coagulations. Irradiation time was 5, 15 and 30 sec. Irradiated energy was 200 mj. After laser irradiation with 5 sec, the absorption coefficient spectrum from 350 nm to 1000 nm did not change without the absorption coefficient peak derived from Hb Fig. 4. The optical properties of the chicken breast tissues after Er:YAG laser coagulations. Irradiation time is 5, 15 and 30 sec. Irradiated energy is 200 mj. (a) absorption coefficient spectra, (b) reduced scattering spectra. Proc. of SPIE Vol F-5

6 observed at 435 nm. The absorption coefficient peak from 435 nm to 450 nm slightly decreased after the laser irradiation with 5 sec. After laser irradiation with 15 sec and 30 sec, the absorption coefficient spectra from 350 nm to 1000 nm decreased. Figure 4(b) showed the reduced scattering coefficient spectra of the chicken breast tissues after Er:YAG laser coagulations. Irradiation time is 5, 15 and 30 sec. Irradiated energy was 200 mj. After laser irradiation with 5 sec, the reduced scattering coefficient spectrum from 350 nm to 1000 nm did not change. After laser irradiations with 15 sec and 30 sec, the reduced scattering coefficient spectra from 440 nm to 1000 nm increased with irradiation time. Decreasing points in the reduced scattering coefficients spectra were found at 440 nm, respectively. Some laser coagulation treatments have conducted in ophthalmology, dentistry or tumor therapy and so on. The disadvantage of all in situ coagulation techniques are excessive coagulation caused by visual controlling. This is due to the lack of suitable real-time monitoring techniques which can detect the morphological conditions immediately. Thus, it is necessary to have precise knowledge about light distribution and the resulting thermal coagulation volumes. These factors are consequently influenced by the irradiation parameters of irradiated laser and the optical properties of laser treated tissues. The optical properties obtained in vitro using the combination of double integrating sphere measurements and IAD technique are clearly useful for in vivo applications. The optical properties of the tissues treated by Er:YAG laser changed dramatically after coagulation. This is an important factor, which should be carefully considered in laser treatment planning because these changes can significantly influence the resulting laser distribution in a tissue, and consequently the outcome of therapeutic procedure. During the coagulation process, the absorption coefficient of a treated tissue decreased and the reduced scattering coefficient of a treated tissue increase. This agrees with the reports from previous studies of other tissues. Thus, laser coagulation leads to the changes of light penetration depth. It has been reported that coagulation leads to reduce an optical penetration depth due to an increase of scattering and a decrease of absorption [13]. The changes in the light penetration depth should be accounted for while planning the therapeutic procedure for the realization of safe laser treatments. 4. CONCLUSION We examined to determine the optical properties changes of coagulated tissues after the Er:YAG laser irradiations by using double integrating sphere system and IAD method in the wavelength range of nm. After laser irradiations with a variety of irradiation parameters, the absorption coefficient of a treated tissue decreased and the reduced scattering coefficient of a treated tissue increased. In carbonization, observed in 30sec-300 mj irradiation, the absorption coefficient of a treated tissue re-increased and the reduced scattering coefficient of a treated tissue re-decrease. The optical properties obtained in vitro using the combination of double integrating sphere measurements and IAD technique are useful for in-vivo applications. REFERENCES [1] [2] [3] [4] [5] Pickering, J. W., Prahl, S. A., Wieringen, N. v., Beek, J. F., Sterenborg, H. J. C. M., Gemert, M. J. C. v., Doubleintegrating-sphere system for measuring the optical properties of tissue, Appl. Opt. 32(4), (1993). Vries, G. d., Beek, J. F., Lucassen, G. W., Gemert, M. J. C. v., The effect of light losses in double integrating spheres on optical properties estimation, IEEE J. Select. Top. Quantum Electron. 5(4), (1993). Roggan, A., Friebel, M., Dorschel, K., Hahn, A., Muller, G., Optical properties of circulating human blood in the wavelength range nm, J. Biomed. Opt. 4(1), (1999). Troy, T. L., Thennadil, S. N., Optical properties of human skin in the near infrared wavelength range of 1000 to 2200 nm, J. Biomed. Opt. 6(2), (2001). Wei, H.-J., Xing, D., Wu, G.-Y., Gu, H.-M., Lu, J.-J., Jin, Y., Li, X.-Y., Differences in optical properties between healthy and pathological human colon tissues using a Ti:sapphire laser: an in vitro study using the Monte Carlo inversion technique, J. Biomed. Opt 10(4), ;1-8 (2005). Proc. of SPIE Vol F-6

7 [6] [7] [8] [9] [10] [11] [12] [13] [14] Meinke, M., Muller, G., Helfmann, J., Friebel, M., Optical properties of platelets and blood plasma and their influence on the optical behavior of whole blood in the visible to near infrared wavelength range, J. Biomed. Opt. 12(1), ;1-9 (2007). Friebel, M., Roggan, A., Muller, G., Meinke, M., Determination of optical properties of human blood in the spectral range nm using Monte Carlo simulations with hematocrit-dependent effective scattering phase functions, J. Biomed. Opt. 11(3), ;1-10 (2006). Salomatina, E., Jiang, B., Novak, J., Yaroslavsky, A. N., Optical properties of normal and cancerous human skin in the visible and near-infrared spectral range, J. Biomed. Opt. 11(6), ;1-9 (2006). Prahl, S. A., Gemert, M. J. C. v., Welch, A. J., Determining the optical properties of turbid media by using the adding-doubling method, Appl. Opt. 32(4), (1993). Prahl, S. A., The Oregon Medical Laser Center at Providence St. Vincent Medical Center, Gebhart, S. C., Lin, W. C., Mahadevan-Jansen, A., In vitro determination of normal and neoplastic human brain tissue optical properties using inverse adding-doubling, Phys. Med. Biol. 51, (2006). Ritz, J.-P., Roggan, A., Isbert, C., Muller, G., Buhr, H. J., Germer, C.-T., Optical properties of native and coagulated porcine liver tissue between 400 and 2400 nm, Lasers Surg. Med. 29, (2001). Yaroslavsky, A. N., Schulze, P. C., Yaroslavsky, I. V., Schober, R., Ulrich, F., Schwarzmaier, H.-J., Optical properties of selected native and coagulated human brain tissues in vitro in the visible and near infrared spectral range, Phys. Med. Biol. 47, (2002). J. MORITA MFG. CORP., or Proc. of SPIE Vol F-7

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