Dipartimento di Ingegneria Chimica, Università di Napoli Federico II, Piazzale Tecchio 80, 80125 Napoli, Italy.
Cytometry A. 2012 Dec;81(12):1040-7. doi: 10.1002/cyto.a.22215. Epub 2012 Oct 18.
The red blood cells (RBCs) population is characterized by a high heterogeneity in membrane area, cellular volume, and mechanical properties, mainly due to the variety of mechanical and chemical stresses that a red cell undergoes in its entire life span. Here, we provide the first simultaneous area and volume measurements of RBCs flowing in microcapillaries, by using high-speed video microscopy imaging and quantitative data processing based on image analysis techniques. Both confined and unbounded flow conditions (depending on the relative size of RBCs and microcapillary diameter) are investigated. The results are compared with micropipette experiments from the literature and data from Coulter counter routine clinical blood tests. Good agreement is found for RBC volume, especially in the case of confined flow conditions. Surface area measurements, which are lacking in the routine clinical test, are of special interest being a potential diagnostic parameter of altered cell deformability and aggregability. Overall, our results provide a novel flow methodology suitable for high-throughput measurements of RBC geometrical parameters, allowing one to overcome the limits of classical static methods, such as micropipette aspiration, which are not suitable for handling a large number of cells.
红细胞(RBC)群体的特征是其膜面积、细胞体积和力学性质具有高度异质性,主要归因于红细胞在其整个生命周期中经历的各种机械和化学应力。在这里,我们通过使用高速视频显微镜成像和基于图像分析技术的定量数据处理,首次提供了在微管中流动的 RBC 的面积和体积的同时测量。研究了受限和无约束流动条件(取决于 RBC 和微管直径的相对大小)。将结果与文献中的微管实验和库尔特计数器常规临床血液测试的数据进行了比较。对于 RBC 体积,特别是在受限流动条件下,发现了很好的一致性。作为细胞变形性和聚集性改变的潜在诊断参数,缺少常规临床测试的表面积测量具有特殊意义。总体而言,我们的结果提供了一种新颖的流动方法,适用于 RBC 几何参数的高通量测量,克服了经典静态方法(如不适合处理大量细胞的微管吸吮)的局限性。