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
- - - - -

Cell Fixation for DNA flow cytometry in fibroblasts

propidium iodide ethanol fixation fibroblast cytometry

  • Please log in to reply
2 replies to this topic

#1 joamartinez

joamartinez

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 02 September 2012 - 06:41 PM

I was wondering if someone could help me with my problem.
I've been trying to read my fibroblasts from adventitia in an hemocytometer. I dont know why but my cells dont get any difference. I believe that they destroy when I fixate them with ethanol 70% as every protocol indicates. I use IP as a dyer. I used to starvate them for 24 hours and then stimulate them whit Serum for 16, 18, 20, 22 and 24 hours in order to snapshot different cell cycle times.I f someone has ever worked with this kind of cells and know any tip or special protocol I would be totally gratefull.

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,835 posts
415
Excellent

Posted 03 September 2012 - 02:04 PM

I'm not quite sure what you are asking - but yes, ethanol will kill the cells, so that a trypan blue stain will only show dead cells after fixation.

It is possible to do live cell FACS, but I don't know any thing about it.

#3 leelee

leelee

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 652 posts
53
Excellent

Posted 03 September 2012 - 06:18 PM

Your topic says flow cytometry, but then you are talking about a hemocytometer? I'm confused, which do you mean?

At any rate, what do you mean you don't get any difference? Any difference in what?





Also tagged with one or more of these keywords: propidium iodide, ethanol, fixation, fibroblast, cytometry

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

©1999-2013 Protocol Online, All rights reserved.