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Calibration-free method to determine the size and hemoglobin concentration of individual red blood cells from light scattering

机译:通过光散射确定单个红细胞的大小和血红蛋白浓度的免校准方法

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At present, hemoglobin concentration and the volume of an erythrocyte can be determined from the intensities of light scattered by an individual cell at fixed angular intervals. This method is used in modern hemoglobin analyzers, but it requires calibration of optical and electronic units by certified particles of known size and refractive index. We describe a method that is based on the parametric solution of an inverse light-scattering problem and does not require a calibration procedure. The method is based on the use of parameters of the entire angular light-scattering pattern, called an indicatrix here. These parameters do not depend on the absolute intensity of light scattering. The indicatrix parameters form approximating equations that relate these parameters to the size and the phase-shift parameters of the particle. The applicability of the method is demonstrated by measurement of the indicatrices of individual sphered erythrocytes. The indicatrices of the individual erythrocytes were measured with a scanning flow cytometer at an angular range of from 15 to 55 deg. The volume and the hemoglobin concentration have been calculated by use of the developed method and by fitting of the experimental indicatrices to the indicatrices calculated from the Mie theory.
机译:目前,血红蛋白浓度和红细胞体积可以由单个细胞以固定的角度间隔散射的光的强度来确定。此方法已用于现代血红蛋白分析仪中,但需要通过已知大小和折射率的认证颗粒对光学和电子单位进行校准。我们描述了一种方法,该方法基于逆光散射问题的参数解,并且不需要校准过程。该方法基于整个角度光散射图案的参数的使用,在此称为indicatrix。这些参数不取决于光散射的绝对强度。指标参数形成将这些参数与粒子的大小和相移参数相关联的近似方程。该方法的适用性通过测量单个球状红细胞的指标来证明。用扫描流式细胞仪在15至55度的角度范围内测量单个红细胞的指标。体积和血红蛋白浓度已经通过使用所开发的方法并且通过将实验指标适合于根据米氏理论计算的指标来计算。

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