Geometrical characterization of healthy red blood cells using digital holographic microscopy and parametric shape models for biophysical studies and diagnostic applications

Modeling of the red blood cell (RBC) shape is an integral part of the experimental and computer simulation investigations of light scattering by these cells for fundamental studies as well as diagnostic applications in techniques like cytometry and quantitative phase imaging. In the present work, a comprehensive study of the geometrical characterization of healthy human RBCs using digital holographic microscopy (DHM) and six frequently employed parametric shape models is reported. It is shown that the comparison of the optical phase profiles, and the thickness profiles given by the models with the DHM results gives a better judgment of the appropriateness of the parametric shape models. It is also shown that the RBC parametric models offer a simpler solution to the refractive index-thickness decoupling problem in QPI methods. Results of geometrical characterization of 500 healthy RBCs in terms of volume, surface area (SA), and sphericity index (SI) led to the classification of the parametric models in two categories based on the nature of variation of these quantities with the cell diameter. In light of the variability of the healthy RBC shapes, our findings suggest that the parametric models exhibiting a negative correlation between the SI and the cell diameter would provide more reliable estimates of the RBC parameters in diagnostic applications. Statistical distributions and descriptive statistics of the RBC volume, SA and SI serve as a guide for the assessment of the capability of the studied parametric models to give a reliable account of the variability of the healthy RBC shape and size.