49
OCTOBER 16-18, 2011 • PORTLAND, OR
P13
Using RPMI Transport Media Increases Recovery
of Paucicellular Specimens Submitted for Flow
Cytometry Testing
Bruce W Greig
1
, Maryalice Stetler-Stevenson
2
1
Vanderbilt University Medical Center, Nashville, TN,
USA,
2
NIH/ NCI, Bethesda, MD, USA
Introduction:
Flow cytometry testing of paucicellular
specimens such as CSF has historically been
challenging due primarily to lack of suffcient cells for
adequate testing purposes. Several fow cytometric
protocols designed to increase cellular yield have
been reported, including immediate stabilization
of cells in Transfx ™ or RPMI with fetal calf
serum (FCS) at the time of specimen collection
1
.
On average, approximately 200 CSF samples
are received annually in our laboratory for fow
cytometric testing. In many cases these specimens
are collected from pediatric patients with possible
CNS involvement with leukemia. We report on
preanalytical steps that improve recovery and results
in a series of CSF specimens.
Methods
: 105 CSF
specimens were studied and results compared to
217 consecutive CSF specimens previously received
in the laboratory. Previous CSF specimens were not
placed in media and were processed using published
CLSI guidelines
2
. Preanalytical procedures were
modifed in that CSF specimens were immediately
placed in RPMI upon receipt in the laboratory and
setup procedures were altered to minimize specimen
manipulation on the premise that “less equals more
cells”.
Results:
Adding RPMI to CSF specimens
and limiting centrifugation to a single step has
greatly increased recovery of viable intact cells for
testing purposes. Using the current method on 105
specimens, 98 (94%) had adequate numbers of viable
cells (>100) suitable for performing fow cytometry
testing.
Conclusions:
.Adding RMPI as a stabilization
media and reducing specimen manipulation results
in increased cell yield and improved fow cytometric
testing results.
POSTER ABSTRACTS