M. Peddi, A., Muthusubramaniam, L., Zheng, Y. F., Cherezov, V., Caffrey,, High-throughput Automated System for Crystallizing Membrane Proteins in Lipidic Mesophases., IEEE Transactions on Automation Science and Engineering, 4, 2, 2007, 129-140
IEEE Transactions on Automation Science and Engineering; 4; 2;
A high-throughput robotic system has been developed
to enable the automatic handling of nanoliter volumes of highly viscous
biomaterials for crystallizing membrane proteins using lipidic
mesophases. The in meso method introduced a few years ago has
produced crystal structures of a number of important membrane
proteins. The bottleneck to achieve high throughput automation
of this, so-called in meso method, is the handling of nanoliter volumes
of the highly viscous cubic phase as part of the crystallization
process. The cubic phase sticks to everything that it has contact
with and has a tendency to disable dispensing robotic tools.
In this paper, we discuss the factors that influence the successful
and automatic delivery of nanoliter volumes of the cubic phase.
A creative cubic-phase-based coordinate measuring mechanism is
presented for controlling the dispensing distance of the cubic phase
which is critically important for the successful performance of the
system. A mathematical model describing the cubic-phase delivery
is proposed and verified.We also present the optimization of liquid
handling parameters for the successful and automatic delivery of
different precipitant solutions. The performance characteristics of
the robotic system in terms of accuracy and reproducibility of delivering
nano volumes of highly viscous biomaterials and micro volumes
of different precipitant solutions are reported.
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