52
ICCS 2011
P16
Rare Event Analysis Using Acoustic Cytometry
Kristi L Haataja, Jolene A Bradford
Life Technologies, Inc., Eugene, OR, USA
Analysis of rare cell populations by fow cytometry
requires the collection of high numbers of events
in order to attain a reliable measure of accuracy,
which leads to long acquisition times using traditional
hydrodynamic focusing. An accurate measurement
of a population with a 1/1,000 probability requires
the collection of 1.6x10
6
events to detect the
population with a CV of 2.5%. When the probability
of event decreases to 1/10,000 the number of events
needed for a CV of 2.5% increases to 1.6x10
7
. The
ability of acoustic cytometry to run samples at up
to 1000 μL/min while maintaining precision and
accuracy provides a distinct advantage over typical
hydrodynamic focusing cytometers for collecting such
large samples. We present two panels which show
the power of acoustic cytometry to make sensitive and
accurate measurements while greatly increasing the
speed to acquire very large sample volumes. First, a
panel was developed to detect mouse plasmacytoid
dendritic cells (pDCs). This specialized cell population
that produces large amounts of type I interferons in
response to viruses, typically comprises less than
0.5% of the total splenocyte population in naïve mice.
A second panel was developed to detect extremely
rare circulating endothelial cells (CECs) which have a
typical range for healthy individuals of 1x10
-7
to 1x10
-5
CEC per leukocyte (1–20 cells/mL of venous blood).
To reduce the risk of loss of CECs in processing, a
no-lyse, no-wash procedure was developed which
requires the speed and accuracy of acoustic focusing
to process such dilute samples.
POSTER ABSTRACTS