Electron Microscope Analysis of Young and Old Red Blood Cells Stained with Colloidal Iron for Surface Charge Evaluation

Human and rabbit red blood cells, separated into "young" and "old" age groups by differential flotation on phthalate esters, were fixed with glutaraldehyde and labeled with colloidal ferric oxide. Electron micrographs of thin sections of young cells showed a uniform and dense depostion of positive iron particles. Old cells showed particles deposited irregularly, leaving unlabeled gaps on the membrane surface. Red cells incubated with 10 units/ml receptor-destroying enzyme (RDE) demonstrate a reduced labeling, similar to that of old cells. After neuraminic acid had been removed from red cells by 20 units/ml RDE, no iron particles were found on membrane surfaces. The different labeling of young, old, and RDE-treated human and rabbit red cells was correlated with their electric mobility and agglutinability by poly-L-lysine. The contradiction between the apparent similarity in charge density of human and rabbit red cells as estimated by density of iron particles and the markedly lower electric mobility of rabbit red cells is discussed.