CYTOMETERS TESTED
MFI has been tested on files from Becton-Dickinson cytometers employing
FACScan, FACStar, LYSYS II, Consort 30, and FACS 440 software, as well as on
files from the Coulter Elite cytometer, and a file generated by Verity
Software's WINLIST. I will attempt to adapt it to files from other sources as
soon as they are supplied to me by interested potential users. The Coulter
Elite Profile produces nonstandard files which require conversion for use with
MFI.
LOG->LINEAR, BLANK SUBTRACTION, PEAK DETECTION, TIME SLICING
MFI calculates median (and mean) fluorescence intensities for
rectangularly-gated events. It automatically converts log-acquired data to a
linear scale, subtracts the "blank" fluorescence intensities for any number of
arbitrarily tagged control files, and converts to kilo-molecules of equivalent
soluble fluorescein if desired. Multiple peaks are detected automatically by
criteria of adjustable sensitivity; medians and event percentages are
automatically calculated for subpopulations. A single data file can be
time-sliced for kinetic analysis.
PRINTABLE OVERLAYED HISTOGRAMS, DOT PLOTS, GATE SETTING
MFI optionally shows histograms for up to 6 parameters and a dot plot on a
single screen. Each graph can be magnified to full-screen if desired.
2-parameter gates can be set graphically; limits for a third parameter (e.g.
propidium iodide) can be entered manually. All graphics screens can be
printed on a variety of printers. Histograms from up to 2 previous files can
be overlayed on those for subsequent files, and can be smoothed. Gated
histograms (optionally smoothed) can also be written as ASCII data files for
plotting by publication-quality software.
EASE OF USE
MFI is completely menu-driven and easy to use. It has extensive built-in
help. It has many configuration options; all configuration selections,
including arbitrarily ordered input filenames, are automatically saved so that
each session begins with the previous settings. Unnecessary parameters (e.g.
FL2 and FL3 when only fluorescein was used) can be omitted from the output,
and parameters can be put out in any order.
List mode data files must be available on a PC compatible computer since MFI
does not operate on HP, VAX or other operating systems. A summary of methods
for transferring files from HP's to PC's is available. The C source code for
MFI is available should someone wish to port it to other operating systems.
HOW TO GET IT
MFI version 3.0, released November 1992, is available by anonymous FTP at
flowcyt.cyto.purdue.edu in directory pub/martz. On request, I will transmit
it UUENCODED by internet, or, if absolutely necessary, snail-mail a floppy
diskette.
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Eric Martz emartz@titan.ucc.umass.edu Professor of Immunology
Voice 413-545-2325, FAX 413-545-1578
Morrill IVN Rm 203, Dept Microbiol, Univ Mass, Amherst MA 01003 USA
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