Light-scattering study of depolymerization kinetics of sickle hemoglobin polymers inside single erythrocytes

Abstract

The time course of intracellular depolymerization of hemoglobin S aggregates is directly observed by using microscope laser light-scattering spectroscopy in single sickle erythrocytes upon slow reoxygenation. From the correlation functions of the light intensity scattered from a single cell, we determine the average diffusion coefficient as well as the fraction of hemoglobin aggregates that have intracellular mobility. The oxygen saturation of the hemoglobin of the same cell is measured by single-cell absorption spectrophotometry. Combining the results obtained with these techniques and information on the cellular morphology, we propose a model for the depolymerization process of hemoglobin inside single erythrocytes as they transform from the fully sickled to the normal biconcave shape upon reoxygenation.