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>Does anyone know of a program we
>can use to calculate mean channel ratio vs time and percent responding cells
>from our stored data?
I use PC-LYSYS. You could also use LYSYS II. Indo-1 "gurus" use MultiTime
from Phoenix Flow Systems. When I looked at MultiTime a while back, it
didn't seem to allow the kind of time "slicing" (gating) I wanted to do, or
it was awkward, or something. You may also want to look at Eric Martz's MFI
program for MS-DOS (MFI = Mean Fluorescense Intensity = mean channel ratio).
I haven't evaluated MFI for Indo-1 yet. Obviously, just about any general
purpose flow software should be able to handle Indo-1 data; it's just a
matter of which software will give you the answers you want with the least
effort and in the least time. I plan on evaluating BDIS's new Paint-a-Gate Pro,
and possibly Attractors, as a replacement to my PC-LYSYS protocol.
>It would also be helpful to know how we might save
>dotplots instead of listmode files on the HP. This would enable us to use our
>Cicero V. 3.10 to make these calculations.
You can indirectly save "dotplots" using LYSYS II. First, save a listmode
file the same as you do now. Second, exit acquistion mode of LYSYS II to
enter analysis mode. Draw a contour plot (NOT a dot plot). Select the "Save
Contour File" item from the Contour menu. I'm not sure that Cicero can read
these files, since I have no experience with Cicero. Also note that contour
files can only be read by LYSYS II as contour plots, NOT dot plots, and
that you will lose all of the listmode correlations. I would recommend
keeping the listmode files "just in case" you need the listmode correlations
(you may want to adjust a FSC/SSC gate, for example).
>We have tried to accumulate the
>data directly in Cicero, however; we are simultaneously collecting our data
>plus a low channel ratio range over time. This noise range is calculated as
>50% of our responding cells in Cicero while the HP graph shows that 100% of
>our cells are responding. I hope this makes sense and someone of you have a
>solution. Thanks.
I'm sorry, you lost me on this last part. If it's important, you can try to
explain it again.
As an aside, you may have noticed that there are numerous published methods
for determining "mean channel ratio" and "percent responding".
/\/\/\_ Eric Van Buren, vanburen%flovax.dnet@rocdec.roc.wayne.edu
\ \ \ Immunology & Microbiology
\_^_/ Wayne State University, Detroit, Michigan
--------------------
Great Lakes International Imaging and Flow Cytometry Association presents
their Fourth Annual Flowdown -- GLIIFCA 4 -- September 29-30, October 1.
Need more information? Call Dr. Alexander Nakeff, (313)577-7923, or FAX him
at (313)577-7925, or (if you're feeling brave), try sending e-mail (not
guaranteed) to nakeff@oncgate.roc.wayne.edu.
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