Fluorescence polarization with FDA in leukaemic cells: a clear difference between myelogenous and lymphocytic origins

Abstract

Intracellular fluorescence polarization (IFP) values of normal human lymphocytes and leukaemic cells from newly diagnosed patients were determined from fluorescence polarization using fluorescein diacetate (FDA). Thirty healthy donors and 40 patients with various types of leukaemia (20 myelogenous and 20 lymphocytic) were included in the present studies. The result was that myeloid cells had about twice the polarization value of lymphocytic cells. The use of FDA for the determination of IFP appears to be useful for differential diagnosis, at least between acute myelogenous and lymphocytic leukaemias. These 2 types of leukaemia also showed a pronounced difference in fluorescence intensity when treated with FDA, perhaps owing to a difference in uptake velocity. The previously described membrane microviscosity using 1,6-diphenyl-1,3,5-hexatriene (DPH), however, did not show such a difference between these 2 leukaemias. The fluorescein-binding protein(s) was also investigated in order to clarify its effect on IFP, but there seemed little evidence for the existence of any such dyebinding protein(s). The advantages of the present method, using FDA, reside in its simplicity, rapidity and considerable sensitivity, requiring a small sample of blood usually less than 5 ml.