Physikalisch-Technische Bundesanstalt (PTB), Abbestraße 2-12, 10587, Berlin, Germany.
Institute of Transfusion Medicine, Charité-Universitätsmedizin Berlin, Charitéplatz 1, 10117, Berlin, Germany.
Sci Rep. 2019 Mar 15;9(1):4623. doi: 10.1038/s41598-019-38767-5.
The knowledge of optical properties of biological cells is essential to interpret their interaction with light and to derive morphological information and parameters associated with cell function like the oxygen transport capacity of human red blood cells (RBCs). We present a method to determine the dependence between the refractive index (RI) of human RBCs and their intracellular hemoglobin (Hb) concentration from spectral extinction measurements of a cell suspension. The procedure is based on the analysis of the corresponding ensemble averaged extinction cross section [Formula: see text]. Thus far two complementary approaches have been taken to derive RIs of RBCs. The first one uses homogeneous macroscopic samples prepared by hemolysis for the destruction of the RBCs' membranes and subsequent centrifugation. A second approach is the determination of RIs of single intact cells by microscopic investigation. These techniques are limited to a few discrete wavelengths or a rather narrow wavelength range. In addition most of these techniques require additional information about the concentration dependence. In contrast, our approach yields the RI increment with Hb concentration of intact, reversibly isovolumetrically sphered, oxygenated RBCs over a wide wavelength range from 290 nm to 1100 nm from macroscopic measurements.
了解生物细胞的光学特性对于解释它们与光的相互作用以及获取与细胞功能相关的形态信息和参数(如人红细胞的氧运输能力)至关重要。我们提出了一种从细胞悬浮液的光谱消光测量中确定人红细胞的折射率(RI)与其细胞内血红蛋白(Hb)浓度之间的依赖性的方法。该过程基于对相应的整体平均消光截面[公式:见文本]的分析。迄今为止,已经采用了两种互补的方法来推导 RBC 的折射率。第一种方法使用通过溶血制备的均匀宏观样品来破坏 RBC 的细胞膜,然后进行离心。第二种方法是通过微观研究确定单个完整细胞的 RI。这些技术仅限于少数离散波长或相当窄的波长范围。此外,这些技术中的大多数都需要有关浓度依赖性的其他信息。相比之下,我们的方法从宏观测量中在从 290nm 到 1100nm 的宽波长范围内产生了完整的、可逆转等容球形的、氧合 RBC 的 Hb 浓度的 RI 增量。