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We have been using To-Pro-3 in a limited way in our Lab. It can be used
for DNA analysis in a similar manner to propidium iodide ie using cells
fixed in ethanol. We use a TP3 concentration of 1uM and get a profile
comparable to PI using 25mW He-Ne excitation. A good reference for its
use is: Van Hooijdonk et al Cytometry (1994), 17, 185-189
Although we havent used it in conjugation with surface staining, as long
as the antigen you are looking at can survive the fixation, there should
be no problem (!).
It is also useful as a dead cell discriminator in two-colour (FITC and
PE) experiments. As people have mentioned in a parallel thread, its use
with third-colour 488nm-excitable tandem dyes may be problematical and if
anyone has used TP3 in this situation, I would be very interested in the
findings!
I should also mention that these experiments have been performed on a
sorter. It should also be possible on the FACS Calibur once the second
laser upgrade is available..........
Derek
Derek Davies
FACS Laboratory,
Imperial Cancer Research Fund, London, UK
http://www.icnet.uk/axp/facs/davies/index.html
On Mon, 23 Sep 1996, Charles Kuszynski wrote:
>
> Dear colleagues,
> I am interested in your experiences using TO-PRO-3 or other DNA
> dyes to do cell cycle analysis using a HENE laser for excitation. This
> needs to be done in conjunction with two cell surface markers on the
> first laser. I would appreciate your kind help
>
>
> Charles Kuszynski
> Univ. Nebr. Med. Cntr
> Omaha, NE 68198-6495
> Voice 402-559-6267
> Fax 402-559-4077
> email ckuszyns@unmc.edu
>
>
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