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
- - - - -

RNA isolation following apoptotic cell labeling


  • Please log in to reply
3 replies to this topic

#1 tmbonney

tmbonney

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 25 February 2009 - 01:05 PM

I am trying to find a way to identify apoptotic cells in a frozen tissue section prior to laser microdissection of NON-apoptotic cells and either protein or RNA isolation. TUNEL wont work as it is destructive to proteins and RNA.

Does anyone have any ideas?

Thanks in advance.

#2 gfischer

gfischer

    Veteran

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 195 posts
9
Neutral

Posted 09 April 2009 - 07:51 AM

I am trying to find a way to identify apoptotic cells in a frozen tissue section prior to laser microdissection of NON-apoptotic cells and either protein or RNA isolation. TUNEL wont work as it is destructive to proteins and RNA.

Does anyone have any ideas?

Thanks in advance.


Does it have to be done in tissue sections? If you can dissociate the tissue, you could run it through FACS and then do RNA iso from the selected population. I know FACS can be used to identify apoptotic and non-apoptotic cells, but I'm not sure how. Try the Flow Cytometry forum if this is an option.
Above all things, if kindness is your king,
then heaven will be yours, before you meet your end

#3 Carlton H

Carlton H

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 95 posts
0
Neutral

Posted 10 April 2009 - 07:31 AM

In many cell types, you can identify apoptotic cells visually. You'll see more "vesicle"-looking bubbles in dying cells. It's particularly easy if a decent percentage of your cell population is apoptotic. If your tissue is transparent and you have a good light microscope, this should work just fine.

Apoptotic cells should also have condensed nuclei. This can be visualized using DAPI or Hoechst. Acridine orange can also be used. Acridine orange stains red when bound to RNA, but green when it binds to DNA. However, it can not enter a normal nuclei. Dying cells (or mitotic cells, but you should be able to tell the difference) will have permeablized nuclei, and the acridine will be able to get in and these cells will fluoresce green while non-mitotic viable cells will not.

-Carlton
NextAdvance
Reliable laboratory instruments for the life sciences, designed to enable you to work more effectively and increase productivity. Check us out!

If my answers help you, please take a moment to check out our site and see if any of our products would help you as well! Thanks!

#4 Anna Lee

Anna Lee

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 24 November 2009 - 08:37 PM

I am trying to find a way to identify apoptotic cells in a frozen tissue section prior to laser microdissection of NON-apoptotic cells and either protein or RNA isolation. TUNEL wont work as it is destructive to proteins and RNA.

Does anyone have any ideas?

Thanks in advance.


Hi all, I am a new member of forum. Would a newcomer be warmly welcome here? Good day you guys!!!
__________________
Watch Anime Episodes




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

©1999-2013 Protocol Online, All rights reserved.