Biomedical Engineering Reference
In-Depth Information
opportunities for therapeutic intervention. As a result, there is an increasing recogni-
tion of the need to progress the “flow of cytometry into systems biology” [18],
a common aim being to bridge the molecular-cellular systems gap [19-21]. Here,
both instrumentation and knowledge from data integration issues present significant
challenges, but if solved, will move systems cytometry to an interface with wider and
more complex systems biology frameworks.
The aim of this chapter is to emphasize the context for flow cytometry in DD&D,
recent advances in the instrumentation offered, new cytometric technologies,
a selective view of some new reagents, and the data handling challenges associated
with cytometry platforms. Throughout we have attempted to reference exemplar
applications that particularly apply to DD&D.
2.2 CURRENT INSTRUMENTATION: ADVANCES IN FLEXIBILITY,
PERFORMANCE, AND THROUGHPUT
A major characteristic of flow cytometry is its inherent versatility as a platform,
allowing refinement for the demands of different stages of drug discovery. Parallel
advances have occurred in the range of lasers available [22], providing new excitation
options for fluors and reducing cost and operation overheads. There has been a trend
over the past decade for flow cytometer manufacturers to move toward digital
electronics with the principal advantages of accurate logarithmic conversions,
convenient compensation matrices, and event triggering on any parameter. Designs
attempt to reduce the disadvantages of digital electronics in the possible loss of
resolution at the low end of the scale, aliasing, and digital pulse processing. The
applications of flow cytometry are also being extended beyond cell profiling into
engineered particle technologies and the analysis of molecular level event detec-
tion [23]. Flow-based approaches are seen as complementary to imagingmethods [24]
or even combined, as in imaging and laser scanning cytometers [25, 26]. Current
instrumentation therefore offers DD&D a range of options for analysis and cell sorting
(Table 2.1). Instruments often share similar performance characteristics for many
applications in DD&D but offer different routes for integration and ease of access.
Here we review selected features of some current platforms to provide a sense of the
range platforms offered.
Significant problems for cell sorting uptake are the operational burden of the
instrumentation setup and the reliability of high-speed preparative performance.
There has been a move toward making cell sorting easier for biologists across a range
of applications. Becton Dickinson Biosciences recently introduced their BD
FACSAria
II flow cytometry system based on a true fixed alignment flow cell that
is gel coupled to the collection optics addressing the need to minimize start-up time.
BD
intowider
availability, offering an open, configurable cell sorting platform with particular
performance characteristics advantageous for small particle detection. BD Flow
Cytometry Analyzers
s recent acquisition of Cytopeia also brings the cell sorting BD Influx
include BD FACSArray
, BD FACSCalibur
,BD
FACSCanto
II, BD LSR
II, and BD LSRFortessa
. Instrument innovations
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