Merino-Dugay A, Cardot P J, Czok M, Guernet M, Andreux J P
Laboratoire de Chimie Analytique et d'Electrochimie Organique, Université Paris-Sud, Châtenay-Malabry, France.
J Chromatogr. 1992 Aug 7;579(1):73-83. doi: 10.1016/0378-4347(92)80364-v.
Gravitational field-flow fractionation is a simple method suitable for the separation of micrometre-size particles, for example the red blood cells (RBC). The variation in the composition of the RBC population in a rabbit during an experimental reversible anaemia induced by phenylhydrazine was studied. Blood samples taken at different stages of the anemia showed differences in the retention and shape of the elution profiles. Microscopic observations of original RBC samples and fractions collected at the outlet of the fractionation channel make a description of the RBC population possible. The three species observed were the normal RBC, the newly produced reticulocytes, and the blood cells containing Heinz bodies (intracellular haemoglobin precipitates). A decrease of the normal RBC from 96 to 1% was observed over five days. The production of reticulocytes in bone marrow is stimulated by the anaemia and increases in percentage after the second day of the anaemia (from 1 to 16%). RBC with Heinz bodies, which appear on the third day, were also studied. Granulometric studies were performed on the RBC sampled from the rabbit each day as well as on some fractions eluted by field-flow fractionation. Reinjection procedures of some cell subpopulations of known size distribution were also performed. The relaxation process of these cells was then studied to approximate their density properties. It was observed that RBC of different density but of the same average size were selectively eluted, as were cells of equivalent density but of different size. Injection of the cells at different stop-flow times enabled the study of the relaxation process on the elution profiles. The results, compared with systematic microscopic observation and size analysis, permit the description of modifications in the RBC composition as well as the purification of subpopulations at each stage of the anaemia. The correlation observed between the fractionation profiles and the progress or the regression of the anaemia opens a new field in the analytical monitoring of this type of pathology.
重力场流分级分离是一种适用于分离微米级颗粒的简单方法，例如红细胞（RBC）。研究了在苯肼诱导的实验性可逆性贫血期间家兔红细胞群体组成的变化。在贫血不同阶段采集的血样显示出洗脱曲线在保留时间和形状上的差异。对原始红细胞样本以及在分级分离通道出口处收集的级分进行显微镜观察，使得描述红细胞群体成为可能。观察到的三种类型分别是正常红细胞、新产生的网织红细胞以及含有海因茨小体（细胞内血红蛋白沉淀）的血细胞。在五天内观察到正常红细胞从96%降至1%。贫血刺激骨髓中网织红细胞的产生，并且在贫血第二天后其百分比增加（从1%增至16%）。还对第三天出现的含有海因茨小体的红细胞进行了研究。对每天从家兔采集的红细胞以及通过场流分级分离洗脱的一些级分进行了粒度分析。还对一些已知大小分布的细胞亚群进行了再注射程序。然后研究了这些细胞的弛豫过程以估算它们的密度特性。观察到不同密度但平均大小相同的红细胞被选择性洗脱，密度相同但大小不同的细胞也是如此。在不同的停流时间注射细胞能够研究洗脱曲线上的弛豫过程。与系统的显微镜观察和大小分析相比，这些结果使得能够描述贫血各阶段红细胞组成的变化以及亚群的纯化。在分级分离曲线与贫血的进展或消退之间观察到的相关性为这类病理学的分析监测开辟了一个新领域。
