Bad FACS data presentation -- more discussion points.

From: Mario Roederer (roederer@drmr.com)
Date: Thu Nov 01 2001 - 14:22:46 EST


I must respectfully disagree with the recent prevailing sentiment
expressed by "old-timers" on this board.  (Since I'm not an
old-timer, and I've been in the field for almost 20 years, then I
guess the definition of old-timers are those who were publishing FACS
data in the 70's).  While my comments below may rankle a few
people--nothing new for me--please be assured that I mean no
disrespect to anyone.

This is a very serious topic, one that has obviously raised a lot of
interest if not emotions in the community.  It is one about which I
have a very central interest, have studied extensively, and have even
taught about.  Hence, I have much to say on the topic.  (What a
surprise.)

First of all, I strongly support an endeavor to produce a document
(or two) to help people present their flow data.  This has been
strongly needed for a long time, and I will be more than happy to
help in any way I can.

But to think that such a document will make a dent in bad data
publications is just plain fantasy.  Face it, the "experts" have been
around for 30 years trying to explain to scientists how to present
data, and it hasn't really helped.  A document such as is proposed
isn't going to be considered outside of our little group of
cytometrists.

Why?  Well, for example, how many people at whom this is targeted
have been bothered to read Howard's or Alice's or anyone else's book
about Flow Cytometry?  If they haven't read those excellent texts (or
even parts of them), I don't think that presenting them with even a
brief missive is going to budge them.

The fact is, scientists don't have the time to assimilate any more
"helpful" documents.  The problem is not that they don't want to, it
is that they don't believe that they have a problem.  They don't
understand what's wrong with what they are doing, especially since
they've been doing it for decades.  And no amount of our going to
them and saying "Hey, here's a better way to present your data" is
going to change them.  Scientists are the ultimate curmudgeons.

I've heard from dozens of Operators or facility Managers that they
try very hard to be a part of the review process for data
presentation out of their facilities.  However, most scientists at
these facilities simply disdain the effort.  What happens?  Our
facility people go to them to help them, but the scientists think
they already know how.  They simply won't accord the facility people
with enough respect, and admitting that maybe the flowjocks might
actually know something the vaunted immunologist doesn't.  Giving the
flowjock a piece of paper to hand the scientist isn't going to be of
much help in this regard.

And to those of you who suggested that we need to do our part by
reviewing the documents that come out of our facilities or our
departments and to provide presentation help to the scientists, you
are 100% correct.  But unfortunately this too is fantasy.  The
reality is, I have enough trouble in my own institute "educating"
people on flow data presentation.  And I have the advantage of being
one of their peers (i.e., "faculty").  They wouldn't give me the time
of day if I were "only" an Operator, despite the fact that most of
the Operators around know far more about flow cytometry than they do.

This is why there has to be an effort to demonstrate what's wrong
with the existing data presentations.  Among others, Zbigniew,
Guenter, Paul, and Howard have been vocal about this being the wrong
approach.  The tenet underlying their opposition is that we don't
want to be negative, we want to be positive about things.  Of course,
this presumes that scientists are a sensitive lot that can't take
criticism and will respond poorly to such criticism.  (Well, OK, many
of them are.  But the majority, I believe, are not.  In fact, I
believe that the majority of scientists would take the criticism in
stride.  To say otherwise would be to accuse them of insecurity.)

The only way to convince scientists to turn to the Operators, to the
Facility Managers, to the flowjocks, is to prove to them that there
are serious problems with what the scientists are publishing.  I
don't like being negative, but I'm willing to bet that for the time
being, it's going to be the only way to get the attention of the
people outside our community.

Of course, once we have their attention, once we've demonstrated that
there are serious flaws in most presentations, then they will turn to
the Facility people and ask for advice.  Then they will be receptive
to documents that illustrate good presentation practices.  Then they
will turn to "only" Operators and discover that the facility people
actually do know what they're talking about.  And maybe they'll start
listening to the Operators, to the Managers.

I am convinced that it is only by empowering the people who actually
run the facilities to educate the scientists, that we will we be able
to change the prevailing presentation techniques.  Only by empowering
the Operators and the Managers to advise the scientists in a
meaningful way will result in long-term success.

This requires a two-pronged approach. First, we must indeed generate
the appropriate educational material -- documents -- that illustrate
what the right way is to present flow data, and why, and how, etc.
This document can have the brief form useful as a checklist for
publication, and as a more complete form that not only gives the
checklist and guidelines, but explains WHY those guidelines are
important.

But second, we must prove that the existing literature is replete
with awful FACS data.  In my opinion, the best way to do this is to
convince editorial boards to solicit a separate specialized review of
all FACS data in tentatively-accepted papers to ensure that the data
is presented properly (and, if there are problems pointed out in the
FACS review, to give the original reviewers a chance to re-evaluate
based on what they missed in the first go-around).  However, I am
sure that editorial boards will be loathe to load up the review
process any more than it already is.  Should we encounter resistance
to this kind of review assistance, then I will stand by my original
suggestion of post-publication review.  Sure, it's ugly, and will
likely be viewed as arrogant.  So what!  Everyone (including
scientists) already thinks that scientists are arrogant.

But it just might have a positive impact:  imagine your local
immunologist finding out that the data presentation atrocities he or
she just published in Nature have been pointed out for the world to
see.  Do you think said immunologist will not think once or twice
about asking for advice with the next paper?  And the bottom line is
that most scientists will admit that the artifacts inherent to the
technology are too many, and often too subtle, for them to be facile
with those problems.  Hence, a forum for those who are familiar with
the problems to illustrate them can be very valuable.  This is
nothing unique to flow cytometry; the recent burgeoning field of
gene-chip analysis has many problems that could be similarly
addressed.

I will admit that this is a radical approach, one that is fraught
with danger because of the perception that it is criticism only for
the point of being critical.  In fact, it is what I consider a
last-ditch effort.  But we've now had 25+ years of bad data.  Just
when is it that we've come to the last ditch, if not now?

mr

(PS:   I found an old "questions" list that I gave my class some
years ago--questions to ask when you are reading a FACS-based
paper--and to kick things off, I put it at
<http://www.drmr.com/facsquestions.html>.  It's very short, it hasn't
been updated in a long time, but it's still valid...)



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