Validation and application of an automated rheoscope for measuring red blood cell deformability distributions in different species

BACKGROUND: The deformability of red blood cells (RBCs) is of great importance for the conservation of oxygen delivery in the microcirculation. Even a small fraction of rigid cells is considered to harm the exchange of respiratory gases. Techniques that measure RBC deformability often provide an indication of the mean deformability. It may not be possible, however, to assess whether this mean value is reduced by the presence of a small rigid cell fraction or by a slight overall reduction in RBC deformability. A technique that provides a deformability distribution would be of great value to study diseases that are marked by subpopulations with a reduced deformability. METHODS: This paper describes a rheoscope system that uses advanced image analysis techniques to quickly quantify the deformability of many individual cells in shear flow, in order to find the RBC-deformability distribution. Since variations in the shear stress are responsible for variations in cell elongation, and hence introduce an additional spread in the cell deformability distribution, we first determined the spread caused by instrumental error. We then utilized the technique to investigate the relation between cell deformability and cell size of single blood samples of different species (human, pig, rat and rabbit). RESULTS: The spread caused by instrumental error was small compared to the actual RBC-deformability spread in blood samples. The deformability distribution of human and pig cells are alike although their cell sizes are different. Rat and rabbit cells show comparable deformability and size distributions. With this technique no correlation was found between cell deformability and cell size in animal RBCs. In the human sample a minor correlation was found between cell deformability and cell size. CONCLUSIONS: The automated rheoscope enables us to study the mechanical properties of RBCs more thoroughly by their deformability distribution. These deformability distributions are hardly influenced by the technique or by cell size.