New PC FACS Software FREE

ERIC MARTZ (emartz@titan.ucc.umass.edu)
Fri, 18 Sep 92 9:46:17 EDT

A new program MFI for calculating Median Fluorescence Intensities on the PC is
offered free of charge for nonprofit research use. The program requires Flow
Cytometry Standard FCS1.0 (or FCS2.0 tho yet untested) list mode data files
such as those generated by FACScan/FACStar. The files must be available on an
IBM-PC compatible computer (can be transferred from HP). To try MFI.EXE,
download MARTZEXE.ARC by FTP from flowcyt.cyto.purdue.edu; it is located in
directory pub/martz.

MFI has no graphics. Rather, it tabulates results compactly, allowing the
results from dozens of files to be summarized on a few pages. The
calculations require only a few seconds per file; hence, results can be
obtained much faster than from the relatively cumbersome FACScan Research
Software. MFI avoids human recording error and warns when crucial parameters
may have been changed accidentally between files. MFI prints the crucial
instrument parameters (gains, threshold, etc.) at the top of a run. If any of
these change between files, a warning message is emitted together with the new
values.

MFI supports rectangular gating (gate limits are best determined by dot plot
examination e.g. in B-D supplied software). To warn of shifts of the event
(dot) cloud within the gate (in the absence of graphic displays), it tabulates
the fraction of events in the upper half and right half of the gate. It can
save several gates at once, and can gate on up to 3 parameters (e.g. scatter
plus propidium iodide).

MFI automatically detects the presence of multiple peaks in histograms (but
ignores the pile-ups which may occur at the ends of the scale). When multiple
peaks are detected, results are tabulated for each peak separately, including
the percentage of the in-gate events in the peak. Peak detection criteria
can be adjusted if desired. Presently, MFI does not provide quadrant
statistics for double-label work (perhaps someone out there would like to add
this capability). It provides peak values one parameter at a time.

MFI tabulates median, mean, mean/median, and converts log-acquired values to a
linear scale or to kilo-molecules of equivalent soluble fluorescein. High and
low values are summarized at the end of a run.

MFI is convenient to use. It has many configuration options, all selected
from menus. It detects FCS files automatically (you don't even have to type
the filename), and allows all or a subset (numeric range) to be processed in a
single run. It saves all data file and configuration choices so that
subsequent runs require few changes in options. MFI has extensive built-in
help/documentation which can be displayed or printed.

The menu system is designed to be simple for programmers to expand or modify.
It uses none of: clear-screen, cursor addressing, colors, bold, reverse, or
ANSI.SYS. Thus, it has its own peculiar but perfectly functional look and
feel, and will be compatible with any operating environment and is eminently
portable to other PC programs or operating systems supporting the C language.
Source code is available on request. The menu/configuration system could be
adapted to other applications.

Future enhancements may include some graphics capabilities (using the
Quinn-Curtis libraries), improvements in peak detection, quadrant/octant
statistics, etc. If anyone wishes to assist in developing enhancements (C
language), please contact the author.

-- 
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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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CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories and distributed free of charge as an educational service to the cytometry community. If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL, Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu EMAIL robinson@flowcyt.cyto.purdue.edu