Microspectrophotometric Determination of Differentially Extracted Haemoglobin in Single Erythrocytes

Abstract

A microspectrophotometric procedure for the determination of insoluble Hb in single cells is presented. Calibration of the method has been carried out on data furnished by the alkali denaturation procedure. The method is applicable to the determination of insoluble Hb and its cellular distribution when the relative concentration of fetal Hb ranges from 10-80% of the total Hb. The results of the present investigation indicate that synthesis of fetal and adult Hb may take place in the same erythroid cell. It is assumed that this microspectrophotometric method is applicable to the determination of fetal Hb in various hemoglobinopathies, provided that optimal conditions and proper calibration of the method are rigorously controlled, also with respect to possible variations in the affinity of the Hb to the cellular stroma which might occur in pathological states.