Mitochondria and Cell Cycle

Jayaraman, Sundararajan anue2468 at uic.edu
Mon Nov 21 10:39:08 EST 2005


I have a paper in press in the Journal of Immunological Methods which is
available online since September 29, 2005. This paper deals with the
ability of TMRE, H2-CMX-Ros and Mito Tracker Red 580 to measure
mitochondrial membrane polarization. I found that TMRE is better
suitable for inner mitochondrial membrane potential measurement by flow
Cytometry in T lymphocytic and in insulin-producing beta cell lines. I
have also analyzed the fixability of these dyes. Both H2-CMX-Ros and Red
580 may be suitable for confocal microscopy after fixation. 
Hope this information will be of help.


With best regards, 

Sundararajan Jayaraman, Ph.D.
Associate Professor
Dept. of Surgery
University of Illinois College of Medicine
909 South Wolcott Avenue
Chicago, IL 60612
Phone: 312 355-5133 (Office); 312-413-0366 (Lab)
Fax:: 312-355-1497


-----Original Message-----
From: McCoy, J. Philip (NIH/NHLBI) [E] [mailto:mccoyj at nhlbi.nih.gov] 
Sent: Friday, November 18, 2005 9:27 AM
To: cyto-inbox
Subject: RE: Mitochondria and Cell Cycle

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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