Re: Set up for Hoechst cell cycle analysis

From: Joseph Webster (J.Webster@centenary.usyd.edu.AU)
Date: Tue Jun 13 2000 - 20:07:29 EST


One thing you probably should check out is the stability of
your laser.
A noisy laser could make for very messy CVs, and I believe
some I-70 lasers do have a problem in certain conditions.

Regards, Joseph.

At 20:46 12/06/2000, Natasha Webb wrote:
>Dear Flow group,
>I am currently having problems setting up a FACS Vantage for Hoechst Cell
>cycle analysis.  It has an Innova 70 laser with UV maximum at 50mW.  I
>would like to know what beads or set up methods others have used to
>optimise the settings to get tight CV's.  I have tried using UV beads from
>Molecular Probes and then Calf thymus nuclei from the BD DNA QC particles
>box with 5 ug/ml Hoechst, however, I cannot get any decent looking cell
>cycle phases - I can only presume there are better methods than this
>attempt.  If anyone has any suggestions, it would be much appreciated.
>Regards
>Natasha.



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