Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

what's that small peak prior to G1?


  • Please log in to reply
2 replies to this topic

#1 zienpiggie

zienpiggie

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 139 posts
3
Neutral

Posted 24 November 2011 - 03:53 PM

Hi all,

I am just wondering if anyone of you have ever seen such a cell cycle pattern like this before for a mammalian cell line. This is untreated cells so I only expect to see a G1 and G2/M peak but I got an extra peak that I've never seen before in another batch of the same cell line or different cell line. Any opinion is hugely appreciated.

Attached Files

  • Attached File  flow.pdf   61.32KB   214 downloads


#2 RynDggn

RynDggn

    member

  • Active Members
  • Pip
  • 14 posts
1
Neutral

Posted 28 November 2011 - 07:36 PM

The peak that is calculated as 52% is the G1 peak (at channel 200). G2/M is then around channel 400. The peak below the G1 is generally referred to as the Sub G1 peak and is indicative of cells that have undergone some DNA degradation due to apoptosis. One way to confirm this would be to display the DNA content on the x-axis and then forward scatter on the y-axis. The Sub G1 cells should be both low in the DNA content axis and low on the FSC channel. For more information on Sub G1, you can see: Kajstura M, Halicka HD, Pryjma J, Darzynkiewicz Z. Discontinuous fragmentation of nuclear DNA during apoptosis revealed by discrete "sub-G1" peaks on DNA content histograms. Cytometry A 2007; 71A:125-131

#3 zienpiggie

zienpiggie

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 139 posts
3
Neutral

Posted 01 December 2011 - 03:16 PM

Hi RynDggn,

thank you very much for the response. This indeed confirmed that the small peak below g1 is apoptotic cells. I wasn't sure because I was using untreated cell lines and was expecting no subg1 peak at all. But the fact that it is present suggests that the cells may be basally apoptotic. I compared this with the same cell line grown in another lab and there was no subg1 peak with theirs.

Thanks again.




Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.