SP Cells

Ian Titley Ian.Titley at icr.ac.uk
Tue Nov 29 06:15:34 EST 2005

One of the original papers on this is Goodell et al J Exp Med 1996
1797-1806 who used 450/20BP, 675 LP and a 610 beam splitter (both
measurements, obviously, from the UV line) on a FACSStarplus.

We have used  a 424/44BP, 670LP and 610 beam splitter eg Jonkers et al
Stem Cells 2005 1059-1065 on a FACSVantageSE (we have also looked since
we had the DiVa upgrade and all is much the same, as one might expect).

So I suspect your blue filter and beam splitter should be OK (I can't
think that 525 or 610 beam splitter would make that much difference in
this assay?) but I would widen things out with a red long pass, 660/20
seems a bit restrictive. Look at both red and blue on linear scales and
the blue intensity should be in the ball park of what you would expect
for a Hoechst stained sample for DNA analysis (red voltage will probably
need to be set a little higher).

Good luck


Ian Titley PhD
Section of Haemato-oncology
Institute of Cancer Research
237 Fulham Road
London UK SW3 6JB

Tel +44 (0)20 7352 8133
Fax +44 (0)20 7352 3299

>>> Leonie Gaudry <L.gaudry at unsw.edu.au> 28/11/2005 05:42:54 >>>

I am having a bit of trouble setting up the Voltages and Compensation
SP Cells.
I am acquiring the cells on a BD FACSVantage with DiVa option in
Digital Mode.
How critical are the filters,I have a Hoechst 33342 450/20 for the blue
a 660/20 for the red but I only have a 525DCSP for the splitter. Are
okay or not specific enough?

I also have a problem in the two papers I am referring too. One paper
states that the detectors should be in Linear mode while a second paper
in Log. Up till now I have been running in Log so which is correct and
would be very interested to hear anyones experiences.
Thank you for your help.
Best Wishes

More information about the Cytometry mailing list