Biomedical Engineering Reference
In-Depth Information
4.6.3 XYZ Minisampler
TheHyperCyt systemuses a peristaltic pump to transport a segment streamof samples
through
30 s and a thin film of
material that coats the tubing from one sample to another. We have developed a
prototype version of a plate transport system to deliver samples directly to the Accuri
C6 flow cytometer. Since this flow cytometer sips sample without a need for
pressurization or external pump, sample wells can be delivered to the sample intake
line. The reduction in path length reduces carryover and transit time. The contents of a
384-well plate have been delivered in less than 20 min.
1m of tubing. This involves a transit time of
4.6.4 Platform Integration for Complex Sample Handling
To our knowledge, the only fully integrated HT flow cytometry system is the Rigel
SiRNA screening system that is dedicated to a highly specialized task involving
repetitive procedures of treating cells with SiRNA, incubating for a particular period
of time, staining the cell as appropriate, and then performing flow cytometric analysis
with HyperCyt. Centrifugation, wash, and resuspension are incorporated as neces-
sary. Because the SiRNA library is considerably smaller than the MLSMR compound
library and because the demands of screening vary dramatically from target to target,
automation is likely to require modular flexibility. Given that overall integration of
sample preparation and flow cytometry has now been demonstrated, the opportunities
for complex sample handling and complex cell suspension are unique to the flow
cytometry platform. From hematopoietic stem cells to blood cells in leukemia,
immunology, inflammation, and infection disease, the possibilities are endless for
HTS flow cytometry based on surface as well as intracellular markers.
4.6.5 Complex Target Data Analysis and Color Coding
Customization of the analysis software for multiplexed targets or complex cell
suspension, and the additional genetic color coding of cells, along with antibody
immunophenotypic staining and conjugated dyes, should compete the overall re-
quirements for HTS flow cytometry and ensure its niche in the discovery arena.
ACKNOWLEDGMENTS
Work at the UNMCMD was supported by NIH Grants U54MH074425 and
U54MH084690. The authors would like to acknowledge the contributions of all the
target providers, the grant support to their projects, and to all team members of the
UNMCMD listed as well as those not listed for their contributions.
REFERENCES
1. Human Genome Project Pilot Phase: http://www.genome.gov/10000511.
2. Human Genome Project Production Phase: http://www.genome.gov/10001792.
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