With a phase microscope the phase shift of cells from type L 929 fibroblast and mitochondria from liver cells was measured. Compared to the total phase shift caused by the cell relative to vacuum (approximately 1400 nm) the single phase shift of the mitochondria (approximately 180 nm) is small. Only the nucleus and the membrane of the cell give a visibly different phase shift relative to the mean value of the cell. The Fraunhofer diffraction of the measured phase object is calculated. With a simplified scattering theory, i.e. Rayleigh-Gans Scattering, different phase objects are investigated and their differential cross section is discussed.
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.
The absorption coefficient, scattering coefficient, and effective scattering phase function of human red blood cells (RBCs) in saline solution were determined for eight different hematocrits (Hcts) between 0.84% and 42.1% in the wavelength range of 250-1100 nm using integrating sphere measurements and inverse Monte Carlo simulation. To allow for biological variability, averaged optical parameters were determined under flow conditions for ten different human blood samples. Based on this standard blood, empirical model functions are presented for the calculation of Hct-dependent optical properties for the RBCs. Changes in the optical properties when saline solution is replaced by blood plasma as the suspension medium were also investigated.
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