Hi everyone!
I've received so many requests for the PDF file -- and many people's
EMail systems won't accept it because of size limitations -- that
I've decided to temporarily post it on the web. I will be removing
it in a week, and there are no direct links to it from any web page,
so you'll have to enter this URL directly into your browser (make
sure you get the capitalization on the letters correct!):
<http://www.drmr.com/BRJIM.pdf>
I am gratified by the significant interest in multi-color work! I
hope everyone who is interested does take Nicole's encouragement to
try it!
mr
(PS, regarding the software compensation issue. Both Rafael and Ray
are incorrect in asserting that I think there's only one package that
does post-hoc compensation. There are quite a few, dating all the
way back to Bob Murphy's CALC4 program in the early 1980's on the
PDP-11 and VAX systems. In fact, compensation is not unique to Flow
Cytometry; scientists have been doing spillover correction
(compensation) for liquid scintillation analysis since before I was
born ("1980").
I would urge Ray and Rafael to re-read what I had written: "there's
really only one package that can handle the complex needs of
multicolor experiments, from off-line compensation of all the
different colors to keeping track of the dozens (if not hundreds) of
distinct gates applied to a single sample."
While many packages can handle compensation, only one can really deal
with the combination of (1) more than 8 parameters; (2) keeping
data-base like track of dozens of gates; (3) automatically applying
compensation matrices to the correct samples; (4) storing the many
different compensation matrices required in a single experiment with
gates, statistics, and analyses; and (5) presenting all of this
information in a manner that can be managed by humans when dealing
with these extremely complex analyses. FlowJo was originally written
and developed at Stanford to deal precisely with the new hurdles &
complexities of 5+ color immunophenotyping experiments; after it was
commercialized by Tree Star, it has continued to evolve so as to
serve a wide segment of the flow community.
Finally, Rafael asks if off-line compensation is necessary for doing
multi-color work. For most 3-color applications, and some 4-color,
the answer is "no". For more than 4 colors, the answer is almost
always yes--and this is because the current generation of hardware is
not sufficiently adept at allowing correct compensation,
easily-defined, and easily-changed on a sample-by-sample basis.)
This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:56:09 EST