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DAB staining and preparation of cultured cells for EM The fixative was removed and cells were washed (five times for 1 min) with 0.1 M sodium cacodylate (Ted Pella, 18851) buffer at pH 7.4 containing 2 mM CaCl 2 on ice and blocked with 20 mM glycine in 0.1 M sodium cacodylate buffer pH 7.4 containing 2 mM CaCl 2 for 20 min on ice

Samples were then analyzed by high-performance liquid chromatography/mass spectrometry in order to determine drug concentrations and calculate negative controls samples containing no liver microsomes [2]

At 7 days of age, ten ducklings were randomly selected from each experimental group, individually weighed, and slaughtered for sample collection