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The cells were then washed (five times for 2 min each) with 0.1 M sodium cacodylate buffer pH 7.4 on ice and were postfixed with 1% osmium tetroxide (EMS, 19150) in 0.1 M sodium cacodylate buffer pH 7.4 for 30 min on ice

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Carefully recap the outer needle cap and twist to remove the used needle