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This work focuses on the use of ultrasound imaging to evaluate the cell concentration of dilute leukocyte suspensions in the range of 10-3000 cells/µL. First, numerical simulations were used to study the influence of the size dispersion and the leukocyte type on the performance of the concentration estimation algorithms, which were developed in previous works assuming single-sized scatterers. From this analysis, corrections to the mentioned algorithms were proposed and then the performance of these corrections was evaluated from experiments. For this, ultrasound images were captured from suspensions of lymphocytes, granulocytes, and their mixtures. These images were obtained using a 20 MHz single-channel scanning system. Results confirmed that concentration estimates provided by conventional algorithms were affected by the size dispersion of cells, leading to a remarkable underestimation of results. The proposed correction to compensate for cell size dispersion obtained from simulations improved the concentration estimation of these algorithms, for the cell suspensions tested, approaching the results to the reference optical characterization. Moreover, it was shown that these models provided a total leukocyte concentration from the ultrasound images which was independent of the relative populations of different white blood cell types.

Evaluation of the Cell Concentration in Suspensions of Human Leukocytes by Ultrasound Imaging: The Influence of Size Dispersion and Cell Type