Beauvoit B, Kitai T, Chance B
Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia 19104.
Biophys J. 1994 Dec;67(6):2501-10. doi: 10.1016/S0006-3495(94)80740-4.
The purpose of this work was to analyze the contribution of the mitochondria to the optical properties, i.e., light absorption and scattering, of the blood-free rat liver. Firstly, a theoretical model of the reduced scattering coefficient of the liver was performed by using the Mie theory, the Rayleigh-Debye-Gans approximation, and the electron microscopy descriptions of the liver ultrastructure. Compared with the hepatocyte volume, the nucleus and the peroxisomes, the mitochondria compartment, accounting for 22% of the liver cell volume, seemed to be the predominant factor for the light scattering of the liver. Second, by using time-resolved spectroscopy and a sample substitution method, we have measured the absorption and reduced scattering coefficients of blood-free perfused rat livers, isolated hepatocyte suspensions, and isolated mitochondria suspensions. A subsequent extrapolation of the isolated mitochondria data to the in vivo mitochondrial content and a comparison with the whole liver measurements lead to the following conclusions: 1) the mitochondria account for about 50% of the liver absorption coefficient at 780 nm (mu a = 0.25 cm-1 extrapolated from isolated mitochondria vs. 0.53 +/- 0.05 cm-1 measured for the liver); and 2) the mitochondrial compartment is the primary factor for the light scattering in the rat liver (mu s' = 15.5 cm-1 extrapolated from the isolated mitochondria versus 15.9 +/- 2.4 cm-1 measured for the liver), demonstrating the relevancy of our preliminary theoretical study.
这项工作的目的是分析线粒体对无血大鼠肝脏光学特性(即光吸收和散射)的贡献。首先,利用米氏理论、瑞利-德拜-甘斯近似以及肝脏超微结构的电子显微镜描述,建立了肝脏约化散射系数的理论模型。与肝细胞体积、细胞核和过氧化物酶体相比,占肝细胞体积22%的线粒体部分似乎是肝脏光散射的主要因素。其次,通过时间分辨光谱和样品替代方法,我们测量了无血灌注大鼠肝脏、分离的肝细胞悬液和分离的线粒体悬液的吸收系数和约化散射系数。随后将分离的线粒体数据外推至体内线粒体含量,并与整个肝脏的测量结果进行比较,得出以下结论:1)线粒体在780 nm处约占肝脏吸收系数的50%(从分离的线粒体外推得到的μa = 0.25 cm-1,而肝脏测量值为0.53±0.05 cm-1);2)线粒体部分是大鼠肝脏光散射的主要因素(从分离的线粒体外推得到的μs' = 15.5 cm-1,而肝脏测量值为15.9±2.4 cm-1),这证明了我们初步理论研究的相关性。