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
- - - - -

Protocol for flow cytometry on mononuclear cells from mouse blood

Flow cytometry mouse

  • Please log in to reply
1 reply to this topic

#1 esbena

esbena

    member

  • Active Members
  • Pip
  • 13 posts
0
Neutral

Posted 26 March 2014 - 03:49 AM

Hallo all.
 
I´m new to flow cytometry and are setting up a protocol for flow cytometry on mononuclear cells from mouse blood, but would like some input from people with experience.
 
I have bought a kit from Cedarlane (Lympholyte®-Mammal Cell Separation Media (Isolation of lymphocytes from Mammalian peripheral blood), sterile liquid) to isolate the mononuclear cells. But I still need some input like: 
 
How much blood is needed to get how much separated cells ?? How many mice are needed ?? How do I collect the blood ?? 
 
I´m in need of a protocol for collection of blood and staining of isolated cells.
 
Thanks


#2 JGBarin

JGBarin

    member

  • Active Members
  • Pip
  • 5 posts
0
Neutral

Posted 11 August 2014 - 09:43 AM

How common/frequent are the cells you're looking for?

 

For relatively common populations (mouse leukocytes are predominantly mononuclear), we've used as little as 50 uL.

 

For very rare populations (say, dendritic cell subsets, or tetramer-positive cells), you may need to essentially exsanguinate the animal via cardiac puncture.


Edited by JGBarin, 11 August 2014 - 09:43 AM.






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

©1999-2013 Protocol Online, All rights reserved.