RECENT ADVANCES IN FLOW CYTOMETRY: PLATFORMS, TOOLS, AND CHALLENGES FOR DATA ANALYSIS - Flow Cytometry in Drug Discovery and Development - page 29

8HT system provides extended cell analysis capabilities using blue (488 nm) and red
(635 nm) excitation lasers and eight detection parameters, including six fluorescent
colors plus forward and side scatter for size and morphology determination. High-
throughput analysis is made possible with a robotic sample tray that can handle a
96-well microplate.
The HyperCyt system, comprising the HyperCyt Autosampler and HyperView
Data Analysis Software, enables existing flow cytometers to analyze samples from
96- or 384-well microplates, with extremely fast rates. Earlier evaluations showed real
promise for enhanced library throughput using a HyperCyt configured platform,
operating with air bubble separated compounds in one sample line and a continuous
stream of cells in another being mixed in-line for serial flow cytometric cell response
analyses [39]. High-throughput flow cytometry (HTFC) using a HyperCyt system has
been used to detect potential anti-inflammatory compounds that block ligand binding
to the formyl peptide receptor (FPR) family of G-protein-coupled receptors (GPCRs)
associated with the localization and activation of tissue-damaging leukocytes at sites
of chronic inflammation [40] InHTSmode, samples were routinely processed at 1.5 s/
well (approximately 2500 cells analyzed per sample), allowing a 96-well plate to be
processed in less than 2.5 min [41]. The S1000 flow cytometry platform from
Stratedigm, Inc. incorporates an architecture that allows up to four lasers to be
mounted on a single plate with solid-state options from 372 to 640 nm. Operationally,
the instrument has a small dead volume of < 8 m L, a minimum sample size of < 20 m L,

and a temperature-controlled 168-tube loader with a bar code reader.
Flow cytometry combined with HCS has the potential to improve biomarker
identification. However, a combination of robotic fluid handling, high-performance
flow cytometric instrumentation, and linked bioinformatics software reveals a limit-
ing step in the processing and analysis of large sample sets in a short period of time:
the degree of automation of the data processing steps [42]. This issue was recently
explored for the extraction of a “cellular signature definition” for acute graft versus
host disease [42].

2.3 ADVANCES IN OPERATING PRINCIPLES: OBJECT
MANIPULATION, SEPARATION, ANALYSIS, AND LIGHT SOURCES

The requirement for cells to be in suspension form for flow cytometric analysis offers
convenient opportunities to modify in-line manipulation of samples and objects of
interest and to integrate analysis of principle of interest in drug development. A typical
example is the extension to flow cytometry of electronic volume measurements, based
on the robust sizing and counting concept introduced by Wallace Coulter (W.H.
Coulter, Means for counting particles suspended in a fluid, U.S. Patent 2,656,508,
priority August 27, 1949). Combination of the technologies allows precise determina-
tion of changes in cell size beyond the resolution provided by the surrogate light
scattering parameters acquired using flowcytometry and provided a basis for initial cell
sorting development [43]. The Cell Lab Quanta SC flow cytometer integrates the

power of electronic sizing with flow cytometry and is highly informative for tracking