Effects of Red Blood Cell Lysing Solutions on the Detection of Peripheral Basophils of Healthy Normals and Sle Patients by Flow Cytometry

Abstract

To evaluate the effect of four widely used red blood cell lysing solutions on counting and measurement of activation marker of peripheral basophils in normals and systemic lupus erythematosus (SLE) patients by flow cytometry. Our results showed that the light scatter properties including FS and SS value of leukocytes in whole blood were preserved when whole blood samples were lysed in RBC Lysis Buffer and FACS Lysing Solution, while were affected when lysed in distilled water or ACK. By counting basophils, RBC Lysis Buffer and FACS Lysing Solution were almost the same level, while were significantly lower when lysed in distilled water or ACK. The expressions of CD203c on peripheral basophils of SLE patients were significantly higher than those of normals. Comparing the data of CD203c expression obtained demonstrated that there were no significant differences among them, while FACS Lysing Solution treatment leads to a slightly lower staining intensity of CD203c. We provide a solid description that the widely used red blood cell lysing reagents may influence the light scatter properties of leukocytes, the accuracy of quantity of absolute number of the existence of basophil subsets and the quantity of staining intensity of cell-activated marker CD203c fluorescence when measured by flow cytometry.