Effects Produced by Anticoagulants or Processing Techniques on RBC Isolation

Cell TypeAdverse Effects
RBC• Oxalate: causes RBCs to shrink or swell, depending on the potassium or ammonium salt used124
 Effect on cell morphology• EDTA: excess of EDTA (>2 mg/mL) causes RBCs to change shape, making morphology difficult to interpret125,126; EDTA at recommended concentrations produces high-quality blood smears if they are made within 2–3 h of the blood draw127
 Effect on cell chemistry• Sodium citrate: RBC morphology unchanged on peripheral blood smear124
 Effect on sample handling or collection• CPD: the levels of lysolecithins (lysoPCs, compounds derived from phosphatidylcholines) are similar between fresh and 5–7-d RBC stored samples106
• Sodium citrate: suitable anticoagulant for routine hematologic analysis on the CELL-DYN 4000 system124
• Oxalate: provides the best RBC labeling compared with ACD, heparin, or sodium oxalate (0.13 μmol/L stannous chloride assay)128
• EDTA: the labeling of RBCs is lower compared with other anticoagulants, likely due to its high chelating capacity128
• Heparin: interferes with Wright's stain124
• RBC number and 2,3-DPG content decrease in whole blood at RTa for 24 h; lactate, potassium, and hemolysis increase, whereas pH, RBC ATP, glucose, and sodium concentrations decrease129
  • ACD, acid citrate dextrose; ATP, adenosine-5′-triphosphate; CPD, citrate-phosphate-dextrose; RBC, red blood cell; 2,3-DPG, 2,3-diphosphoglycerate.

  • a RT = 20–25°C.