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Jeff,
LYSYS II allows several different gating options which provide a high degree
of flexibility. This includes live gating, list gating, display and count
gating. There is a good description of this in the LYSYS II User's manual,
but a quick synopsis on gating options which can effect list data follows.
Live gating, set up by selecting accept or reject on a region, gates data
comming from the cytometer into the LYSYS II program. Any events outside of
an active live gate, in accept mode, are ignored by LYSYS II and thus will
not be saved to a list mode file.
List gating is a "second step" gate, which follows after live gating, which
can filter data sent to the list mode file. List gating can be use with or
without live gating, but in no case can data be saved to a list mode file
that does not satisfy the live gate criteria.
In addition, in the protocol window, LYSYS II allows individual parameters
to be turned on or off, which will also affect what is stored to a list mode
file.
- From the description in your message, it sounds like the data file itself
was not corrupted, since you can see some of the parameters (usually, if a
file is corrupted, either it is rejected completely by the software, or some
of the events across all parameters are either missing or wrong.)
If the problem was in the gating setup of LYSYS II, the data is likely not
there at all to be recovered from the data file. However, I'd recommend you
contact the BD Customer Support Center at (800) 448-BDIS, and they will be
able to analyze the data files to see if there is any corruption.
Mike Lance
Becton Dickinson
(408) 954-2047
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Message-Id: <9310221602.AA05913@sugen1.sugen.sf.ca.us>
Date: Fri, 22 Oct 1993 08:55:22 -0800
To: cytometry@flowcyt.cyto.purdue.edu
From: jeff@sugen.sf.ca.us
Subject: lost data
I have just experienced a problem which I have never faced before. Using
LYSIS II and a BD FACScan, I collected data yesterday and saved it to a
Bernoulli disk. I was using PE-labelled second antibody, and I gated on
live cells using FSC/SSC. I collected 10K events (live gating), and
assumed that I was collecting FSC, SSC, FL1H, FL2H, and FL3H data. In
fact, I know that all my samples were positive for FL2H (except the
negative control), because I could see the data as I ran the samples.
However, today I tried to analyze the data saved on the Bernoulli, and
found to my surprise that only FSC, SSC, and FL1H data were available.
Also, the file size was 61K, where normally 10K events would result in a
file size of 104K. So apparently the FL2H and FL3H data were not saved.
Why not?
Has anyone else experienced this problem? And is there a way to recover my
data or is all hope lost?
******************
* Jeff Powell, Ph.D. *
* SUGEN Inc. *
* Redwood City *
* 415-306-7763 *
******************
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