Knowledge about the optical properties μa,μs, and g of human blood plays an important role for many diagnostic and therapeutic applications in laser medicine and medical diagnostics. They strongly depend on physiological parameters such as oxygen saturation, osmolarity, flow conditions, haematocrit, etc. The integrating sphere technique and inverse Monte Carlo simulations were applied to measure μa,μs, and g of circulating human blood. At 633 nm the optical properties of human blood with a haematocrit of 10% and an oxygen saturation of 98% were found to be 0.210±0.002 mm-1 for μa,77.3±0.5 mm-1 for μs, and 0.994±0.001 for the g factor. An increase of the haematocrit up to 50% lead to a linear increase of absorption and reduced scattering. Variations in osmolarity and wall shear rate led to changes of all three parameters while variations in the oxygen saturation only led to a significant change of the absorption coefficient. A spectrum of all three parameters was measured in the wavelength range 400-2500 nm for oxygenated and deoxygenated blood, showing that blood absorption followed the absorption behavior of haemoglobin and water. The scattering coefficient decreased for wavelengths above 500 nm with approximately λ-1.7; the g factor was higher than 0.9 over the whole wavelength range. © 1999 Society of Photo-Optical Instrumentation Engineers.

The absorption coefficient mu(a), scattering coefficient mu(s), and anisotropy factor g of diluted and undiluted human blood (hematocrit 0.84 and 42.1%) are determined under flow conditions in the wavelength range 250 to 1100 nm, covering the absorption bands of hemoglobin. These values are obtained by high precision integrating sphere measurements in combination with an optimized inverse Monte Carlo simulation (IMCS). With a new algorithm, appropriate effective phase functions could be evaluated for both blood concentrations using the IMCS. The best results are obtained using the Reynolds-McCormick phase function with the variation factor alpha = 1.2 for hematocrit 0.84%, and alpha = 1.7 for hematocrit 42.1%. The obtained data are compared with the parameters given by the Mie theory. The use of IMCS in combination with selected appropriate effective phase functions make it possible to take into account the nonspherical shape of erythrocytes, the phenomenon of coupled absorption and scattering, and multiple scattering and interference phenomena. It is therefore possible for the first time to obtain reasonable results for the optical behavior of human blood, even at high hematocrit and in high hemoglobin absorption areas. Moreover, the limitations of the Mie theory describing the optical properties of blood can be shown.

The real part of the complex refractive index of oxygenated native hemoglobin solutions dependent on concentration was determined in the wavelength range 250 to 1100 nm by Fresnel reflectance measurements. The hemoglobin solution was produced by physical hemolysis of human erythrocytes followed by ultracentrifugation and filtration. A model function is presented for calculating the refractive index of hemoglobin solutions depending on concentration in the wavelength range 250 to 1100 nm.

The intrinsic optical parameters absorption coefficient mu(a), scattering coefficient micros, anisotropy factor g, and effective scattering coefficient micros were determined for human red blood cell (RBC) suspensions of hematocrit 33.2% dependent on the oxygen saturation (SAT O(2)) in the wavelength range 250 to 2,000 nm, including the range above 1,100 nm, about which there are no data available in the literature. Integrating sphere measurements of light transmittance and reflectance in combination with inverse Monte Carlo simulation were carried out for SAT O(2) levels of 100 and 0%. In the wavelength range up to 1,200 nm, the absorption behavior is determined by the hemoglobin absorption. The spectral range above the cells' absorption shows no dependence on SAT O(2) and approximates the absorption of water with values 20 to 30% below the respective values for water. Parameters micros and g are significantly influenced by the SAT O(2)-induced absorption changes. Above 600 nm, micros decreases continuously from values of 85 mm(-1) to values of 30 mm(-1) at 2,000 nm. The anisotropy factor shows a slight decrease with wavelengths above 600 nm. In the spectral regions of 1,450 and 1,900 nm where water has local absorption maxima, g shows a significant decrease down to 0.85, whereas micros increases.

The optical parameters absorption coefficient, scattering coefficient, and the anisotropy factor of platelets (PLTs) suspended in plasma and cell-free blood plasma are determined by measuring the diffuse reflectance, total and diffuse transmission, and subsequently by inverse Monte Carlo simulation. Furthermore, the optical behavior of PLTs and red blood cells suspended in plasma are compared with those suspended in saline solution. Cell-free plasma shows a higher scattering coefficient and anisotropy factor than expected for Rayleigh scattering by plasma proteins. The scattering coefficient of PLTs increases linearly with the PLT concentration. The existence of physiological concentrations of leukocytes has no measurable influence on the absorption and scattering properties of whole blood. In summary, red blood cells predominate over the other blood components by two to three orders of magnitude with regard to absorption and effective scattering. However, substituting saline solution for plasma leads to a significant increase in the effective scattering coefficient and therefore should be taken into consideration.