I have a problem about flow sample preparation. We fix A549 cells with 0.5% PAF for 1 h in 4degC. At the end of fixation, cells were centrifuged with 1200 rpm, 5min, then washed with PBS and centrifuged with same speed. We found that cells were seriously lost (cannot precipitate) after second centrifugation. Can everyone tell us how to change our condition for avoiding sample lose? thanks.
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