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Sorry I'm late chiming in here. I am only now catching up on the listserv
after a few months.
In our live cell article we try to show some test results and in no way try
to offer an extensive test of cell media or reagents. The main point we
always try to make is that things may not behave as you expect and you need
to test it on your system with your cells and your imaging conditions. In
our case Lebowitz does not allow for CHOK1 cell division. Maybe it works
will for other systems?
That being said we did some multiday imaging with Hoechst staining at 2nM in
CHO K1 cells. The cells do not divide in DMEM medium. They try to and often
come back together as one cell. We ended up with very large multi-nucleated
cells with very strange nuclear shapes. That being said in CCM1 and CO2
Independent medium from Invitrogen they will undergo some division but very
reduced when compared to no dye.
In addition cell migraiton rates are reduced by about 50% in all media
tested during the first 24 hours of imaging when compared to cells with no
nuclear probe. We will publish this study as part of a book chapter coming
out in Fluorescence Reviews 2010 in the Fall.
We are gearing up to try a panel of nuclear dyes at various concentrations
in our CHO K1 model cells. I would love to try testing many different cell
types, differnet media and different imaging conditions but we simply don't
have the resources for this. So make sure you test things on your own
systems with your cells, your media and your imaging conditions.
John Sedat and co-workers published a very nice article on light doses in
yeast and affects on cell division even after what appeared to be relatively
low light doses.
Fast live simultaneous multiwavelength four-dimensional optical microscopy.
Carlton PM, Boulanger J, Kervrann C, Sibarita JB, Salamero J, Gordon-Messer
S, Bressan D, Haber JE, Haase S, Shao L, Winoto L, Matsuda A, Kner P, Uzawa
S, Gustafsson M, Kam Z, Agard DA, Sedat JW.
Proc Natl Acad Sci U S A. 2010 Sep 14;107(37):16016-22. Epub 2010 Aug 12.
I think it is worth a read.
Sincerely,
Claire
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