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Cytometry A. 2019 Apr 13. doi: 10.1002/cyto.a.23756. [Epub ahead of print]

Image-Based Flow Cytometry and Angle-Resolved Light Scattering to Define the Sickling Process.

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Department of Electrical and Electronics Engineering, Bilkent University, Ankara 06800, Turkey.
Department of Physiology, School of Medicine, Koc University, Istanbul 34450, Turkey.
Department of Electrical and Computer Engineering, Chair of High-Frequency Engineering, Technical University of Munich, Munich 80333, Germany.
Children's Hospital Oakland Research Institute, Oakland, California, 94609.


Red blood cells (RBCs) from sickle cell patients exposed to a low oxygen tension reveal highly heterogeneous cell morphologies due to the polymerization of sickle hemoglobin (HbS). We show that angle-resolved light scattering approach with the use of image-based flow cytometry provides reliable quantitative data to define the change in morphology of large populations of RBCs from sickle cell patients when the cells are exposed for different times to low oxygen. We characterize the RBC morphological profile by means of a set of morphological and physical parameters, which includes cell shape, size, and orientation. These parameters define the cell as discocyte, sickle, elongated, as well as irregularly or abnormal RBC shaped cells, including echinocytes, holly-leaf, and granular structures. In contrast to microscopy, quick assessment of large numbers of cells provides statistically relevant information of the dynamic process of RBC sickling in time. The use of this approach facilitates the understanding of the processes that define the propensity of sickle blood samples to change their shape, and the ensuing vaso-occlusive events in the circulation of the patients. Moreover, it assists in the evaluation of treatments that include the use of anti-sickling agents, gene therapy-based hemoglobin modifications, as well as other approaches to improve the quality of life of sickle cell patients.


HbS polymerization; computational flow cytometry; light scattering; precision and personalized patient-oriented medicine; red blood cells; sickle cell disease


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