For detailed information, see Kile et al., 2002.
Brief description: Peripheral blood was drawn (150 microliters) by retro-orbital puncture using a Pasteur pipette. Blood was collected in Microtainer brand tubes containing K2EDTA (Becton Dickinson, Franklin Lakes, NJ) and samples analyzed using a Cell-Dyn 3500R automated veterinary hematology analyzer (Abbott Diagnostics, Santa Clara, CA) according to the manufacturer's instructions.
Notes:It has been suggested that complete blood counts (CBCs) obtained by retro-orbital puncture should not differ significantly from counts performed on samples acquired by cardiac puncture (Sluiter et al. 1985). In this study, retro-orbital bleeding gave far more consistent results (see Kile et al., 2002).
Although previous projects have established the veracity of the Cell-Dyn 3000 white blood cell differential counts performed on human peripheral blood (Van Leeuwen et al. 1991; Cornbleet et al. 1992), the accuracy of the Cell-Dyn 3500R veterinary analyzer differentials was confirmed here through manual inspection of peripheral blood smears obtained from male and female C57BL/6J mice.
EDTA is preferable to heparin as an anticoagulant, largely
due to the tendency of heparin-treated blood samples to
undergo platelet aggregation and hence produce false
platelet number and mean platelet volume readouts (Kawamoto
et al. 2000; Muriithi et al. 2000). In addition, heparin is
unsuitable for leukocyte counts or blood smears.