Mitochondria and Cell Cycle

McCoy, J. Philip (NIH/NHLBI) [E] mccoyj at NHLBI.NIH.GOV
Fri Nov 18 10:26:47 EST 2005


We use TMRM or Mitotracker Red (depending on the experiment) and Hoechst
33342.

Phil 

-----Original Message-----
From: Dan Rosson Ph.D. [mailto:Dan.Rosson at mail.jci.tju.edu] 
Sent: Thursday, November 17, 2005 11:50 AM
To: cyto-inbox
Subject: Mitochondria and Cell Cycle

Dear Flowlosophers:

I have a user that has been told by his grant reviewer to measure
mitochondria as a function of cell cycle position. Someone suggested
using Mito Red and YOYO-1. That didn't work out to well for two reasons:
the cell cycle profile looked like garbage and while YOYO has an
emission maximum around 510nM, it is so broad and intense it shows up in
all channels including 675nM where its still impossible to compensate.
The best suggestion I could come up with was to use formaldehyde-fixed
cells with PI for cell cycle and to find an antibody to a mitochondrial
protein (e.g.
cytochrome c) and use FITC to detect that. Does anybody have any better
suggestions?


Dan Rosson PhD
Thomas Jefferson University Flow Jock




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