Staining with PI & BrdU
t-delohery at ski.mskcc.org
Thu Mar 28 01:55:30 EST 1996
>I've been staining cells with BrdU & getting about 60-80% incorporation
>after a 2h pulse - is this the norm? The cycling time of these cells is 12
>h. I've tried adding PI to look at DNA content & find that 48% of the cells
>are in S phase. Do these results make sense? I'm not a DNA person so I'm
>not quite sure. Someone that I spoke to thought that it would make sense,
>but my supervisor doesn't think so. So, I thought I'd ask the opinion of
>people who do this everyday!
Makes perfectly good sense to me. Are thes MEL cells? CHO? ~50% S-phase
with a 12 hr. doubling time tells you these are relatively rapid growers.
Keep in mind the BrdU is labelling ALL cells in S over a 2hr. period - cells
in late S at the beginning of 2h and cells entering S 2hrs later.
This, as opposed to a the % S measured by PI at the instant of fixation.
Thomas Delohery | Internet: t-delohery at ski.mskcc.org
Manager, Flow Cytometry Core Facility | Phone: (212) 639-8729
Memorial Sloan-Kettering Cancer Center | Fax: (212) 794-4019
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