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Operator-assisted harvesting of protein crystals using a universal micromanipulation robot.

Viola R, Carman P, Walsh J, Miller E, Benning M, Frankel D, McPherson A, Cudney B, Rupp B - J Appl Crystallogr (2007)

Bottom Line: High-throughput crystallography has reached a level of automation where complete computer-assisted robotic crystallization pipelines are capable of cocktail preparation, crystallization plate setup, and inspection and interpretation of results.To address the final frontier in achieving fully automated high-throughput crystallography, the prototype of an anthropomorphic six-axis universal micromanipulation robot (UMR) has been designed and tested; this UMR is capable of operator-assisted harvesting and cryoquenching of protein crystals as small as 10 microm from a variety of 96-well plates.The UMR is equipped with a versatile tool exchanger providing full operational flexibility.

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ABSTRACT
High-throughput crystallography has reached a level of automation where complete computer-assisted robotic crystallization pipelines are capable of cocktail preparation, crystallization plate setup, and inspection and interpretation of results. While mounting of crystal pins, data collection and structure solution are highly automated, crystal harvesting and cryocooling remain formidable challenges towards full automation. To address the final frontier in achieving fully automated high-throughput crystallography, the prototype of an anthropomorphic six-axis universal micromanipulation robot (UMR) has been designed and tested; this UMR is capable of operator-assisted harvesting and cryoquenching of protein crystals as small as 10 microm from a variety of 96-well plates. The UMR is equipped with a versatile tool exchanger providing full operational flexibility. Trypsin crystals harvested and cryoquenched using the UMR have yielded a 1.5 A structure demonstrating the feasibility of robotic protein crystal harvesting.

No MeSH data available.


Related in: MedlinePlus

Automated cryoquenching of a crystal in liquid nitrogen. The sample magazine used during the tests was a simplified version of the ‘cryo-pucks’ currently in use at a number of synchrotron light sources. Because of the system’s inherent flexibility, the UMR can be configured for compatibility with any type of storage magazine currently in use.
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fig4: Automated cryoquenching of a crystal in liquid nitrogen. The sample magazine used during the tests was a simplified version of the ‘cryo-pucks’ currently in use at a number of synchrotron light sources. Because of the system’s inherent flexibility, the UMR can be configured for compatibility with any type of storage magazine currently in use.

Mentions: Following successful demonstration of the robot’s ability to perform individual process tasks, system-level tests of the UMR were performed. The goal of these tests was to demonstrate the ability of the UMR to seamlessly perform the sequence of tasks required to completely open the bottleneck associated with the crystal harvesting process. Microtiter plates enter the system through an input port, where the robot accepts the tray and presents it to the imaging module. Magnified images of each of the plate’s wells are acquired and displayed to the system operator. Once the operator identifies those wells that hold usable crystals, the robot precisely cuts through the tape, acquires the appropriately sized harvesting loop and positions the tip of the loop in the center of the well. The operator then takes direct control of the robot and performs the necessary harvesting and cryoprotecting operations. Finally, the operator returns control back to the robot and the robot automatically cryoquenches the crystal (Fig. 4 ▶). During a series of tests with numerous crystals grown in varying plates under different conditions, the feasibility of a tele-operated UMR was established. Ultimately, a crystal structure of trypsin complexed with benzamidine was refined at 1.5 Å from data collected from a robotically harvested and cryoquenched crystal.


Operator-assisted harvesting of protein crystals using a universal micromanipulation robot.

Viola R, Carman P, Walsh J, Miller E, Benning M, Frankel D, McPherson A, Cudney B, Rupp B - J Appl Crystallogr (2007)

Automated cryoquenching of a crystal in liquid nitrogen. The sample magazine used during the tests was a simplified version of the ‘cryo-pucks’ currently in use at a number of synchrotron light sources. Because of the system’s inherent flexibility, the UMR can be configured for compatibility with any type of storage magazine currently in use.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2483483&req=5

fig4: Automated cryoquenching of a crystal in liquid nitrogen. The sample magazine used during the tests was a simplified version of the ‘cryo-pucks’ currently in use at a number of synchrotron light sources. Because of the system’s inherent flexibility, the UMR can be configured for compatibility with any type of storage magazine currently in use.
Mentions: Following successful demonstration of the robot’s ability to perform individual process tasks, system-level tests of the UMR were performed. The goal of these tests was to demonstrate the ability of the UMR to seamlessly perform the sequence of tasks required to completely open the bottleneck associated with the crystal harvesting process. Microtiter plates enter the system through an input port, where the robot accepts the tray and presents it to the imaging module. Magnified images of each of the plate’s wells are acquired and displayed to the system operator. Once the operator identifies those wells that hold usable crystals, the robot precisely cuts through the tape, acquires the appropriately sized harvesting loop and positions the tip of the loop in the center of the well. The operator then takes direct control of the robot and performs the necessary harvesting and cryoprotecting operations. Finally, the operator returns control back to the robot and the robot automatically cryoquenches the crystal (Fig. 4 ▶). During a series of tests with numerous crystals grown in varying plates under different conditions, the feasibility of a tele-operated UMR was established. Ultimately, a crystal structure of trypsin complexed with benzamidine was refined at 1.5 Å from data collected from a robotically harvested and cryoquenched crystal.

Bottom Line: High-throughput crystallography has reached a level of automation where complete computer-assisted robotic crystallization pipelines are capable of cocktail preparation, crystallization plate setup, and inspection and interpretation of results.To address the final frontier in achieving fully automated high-throughput crystallography, the prototype of an anthropomorphic six-axis universal micromanipulation robot (UMR) has been designed and tested; this UMR is capable of operator-assisted harvesting and cryoquenching of protein crystals as small as 10 microm from a variety of 96-well plates.The UMR is equipped with a versatile tool exchanger providing full operational flexibility.

View Article: PubMed Central - HTML - PubMed

ABSTRACT
High-throughput crystallography has reached a level of automation where complete computer-assisted robotic crystallization pipelines are capable of cocktail preparation, crystallization plate setup, and inspection and interpretation of results. While mounting of crystal pins, data collection and structure solution are highly automated, crystal harvesting and cryocooling remain formidable challenges towards full automation. To address the final frontier in achieving fully automated high-throughput crystallography, the prototype of an anthropomorphic six-axis universal micromanipulation robot (UMR) has been designed and tested; this UMR is capable of operator-assisted harvesting and cryoquenching of protein crystals as small as 10 microm from a variety of 96-well plates. The UMR is equipped with a versatile tool exchanger providing full operational flexibility. Trypsin crystals harvested and cryoquenched using the UMR have yielded a 1.5 A structure demonstrating the feasibility of robotic protein crystal harvesting.

No MeSH data available.


Related in: MedlinePlus