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Oxygen is transported throughout the body by hemoglobin in red blood cells.While the oxygen affinity of blood is well understood and is routinely assessedin patients by pulse oximetry, variability at the single-cell level has notbeen previously measured. In contrast, single-cell measurements of red bloodcell volume and hemoglobin concentration are taken millions of times per day byclinical hematology analyzers and are important factors in determining thehealth of the hematologic system. To better understand the variability anddeterminants of oxygen affinity on a cellular level, we have developed a systemthat quantifies the oxygen saturation, cell volume and hemoglobin concentrationfor individual red blood cells in high-throughput. We find that the variabilityin single-cell saturation peaks at an oxygen partial pressure of 2.5%, whichcorresponds to the maximum slope of the oxygen-hemoglobin dissociation curve.In addition, single-cell oxygen affinity is positively correlated withhemoglobin concentration, but independent of osmolarity, which suggestsvariation in the hemoglobin to 2-3 DPG ratio on a cellular level. Byquantifying the functional behavior of a cellular population, our system adds anew dimension to blood cell analysis and other measurements of single-cellvariability.

Single-cell measurement of red blood cell oxygen affinity