Re: Cell cycle, BrdU incorporation, and Apoptosis (TUNEL)

Hi Matthew,
Did you read the discussion on this List titled "Clarification of my Phase of Cell Cycle
at Time of Death
Question" with a number of postings that occured here just two months ago?

I think there has been a lot of information on apoptosis and cell cycle staining on this
List in all years, so as everywhere else.
Regards.
Vladislav Rozenkov
-----Original Message-----
From: Matthew Janes <mjanes@uci.edu>
To: cyto-inbox
Sent: Wed, 21 Jun 2006 12:04:01 -0700
Subject: Cell cycle, BrdU incorporation, and Apoptosis (TUNEL)


I want to determine in what stage of the cell cycle particular primary lymphocytes are
dying. So ask the community if anyone has a recommended protocol for CELL CYCLE (DNA
content with BrdU incorporation to properly distinguish all cell cycle phases) with
TUNEL, or another well established method for assaying for apoptosis. 
Or can sorting of populations in the different stages of the cell cycle be performed? I
am experienced with sorting, so is it a problem sorting different populations when
aquiring data on a linear scale (like when using PI, 7AAD, or DAPI for cell cycle? Would
the community recommend just sorting different lymphocytes based off their cell cycle,
the subsequently enumerate apoptosis by microscopy, or look at the degree of DNA
fragmentation on a gel. 
Any opinion on such a task would be greatly appreciated. 
best, 
Matt 
-- Matthew R. Janes, MSPH 
Ph.D. Candidate 
Department of Molecular Biology & Biochemistry 
Center for Immunology 
University of California at Irvine 
mjanes@uci.edu