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Significance Sickle cell disease is the first “molecular disease” described more than 100 years ago. All morbidity and mortality are ultimately caused by polymerization of the variant sickle hemoglobin molecule in red blood cells (RBCs). However, despite decades of intense research, we are unable to measure the amount of hemoglobin polymer in individual RBCs with high throughput at arbitrary oxygen tension. We describe a high-throughput method to assess hemoglobin polymer at the single-cell level under controlled oxygen tension. The assay infers hemoglobin polymer fraction by measuring its effect on cellular oxygen affinity. This method provides biomarkers promising to improve management of sickle cell patients and to optimize the development and prioritization of candidate cures.

High-throughput assessment of hemoglobin polymer in single red blood cells from sickle cell patients under controlled oxygen tension