[Cytometry] primary mouse hepatocytes cell cycle question

Yoav Altman yoav at sanfordburnham.org
Mon May 9 14:50:55 EDT 2011


Hi Julie,

Mouse hepatocytes can be multinucleated.  We routinely see 2N, 4N, 8N, 16N peaks with mouse hepatocytes.  I'd be inclined to skip manual singlets gating altogether and use a diploid-tetraploid model in your modeling software of choice.  If you have an opportunity to repeat the experiment, the addition of CEN singlets to the mix could help you make sense of the two lower peaks.  

Regards,

Yoav Altman
Director, Flow Cytometry Shared Resource
Sanford-Burnham Medical Research Institute
858-646-3100 x3569
yoav at sanfordburnham.org

10901 North Torrey Pines Road
La Jolla, CA 92037
http://www.sanfordburnham.org



On May 6, 2011, at 12:25 PM, Julie Farnsworth wrote:

> Could someone help me analyze data from primary mouse hepatocytes 
> stained with PI run on BDFACSCanto
> I am not sure where to draw the "singlet gate" for the single cells to 
> exclude doublets and
> not sure what each peak in the histogram represents.
> *What are the two smaller peaks?*
> 
> I hope you can see the dot plots and histograms below.
> Set one shows a larger P1 gate and
> the other set shows a smaller/tighter P1 gate.
> Which one is correct?
> 
> thanks for you help
> 
> 
>> Same data file, 2 different gating P1 regions
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