Minimal intervention robotic protein crystallization

doi:10.1016/0022-0248(91)90879-A

Journal of Crystal Growth

Volume 110, Issues 1–2, 1 March 1991, Pages 156-163

https://doi.org/10.1016/0022-0248(91)90879-A Get rights and content

Abstract

An in-house protein crystallization robotic system has been designed and built to improve the rate for obtaining protein single crystals suitable for X-ray structure determination. The system uses the hanging drop, vapor diffusion technique. Positive features of previous crystallization robot designs in other laboratories were combined with several new features and requirements in the present system. The system is designed to minimize the degree of carry over from one crystallization trial to the next, to operate at 4°C, and to operate sequentially on as many as 51 Linbro 24-well culture dishes without operator intervention. The system has a number of design elements which could be adapted to other robotic crystal growth systems.

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Cited by (14)

[4] Robotics for Automated Crystal Formation and Analysis
2003, Methods in Enzymology
Citation Excerpt :
For example, Ward et al. required 5 μl protein + 5 μl mother liquor per drop,11 whereas the Cox and Weber system used 2 μl protein + 2 μl solution samples per drop.12 Additionally, setup of an entire crystallization trial was slow: 35–40 hr for 51 Linbro 24-well plates to be dispensed (Rubin et al. system13), 15–45 min to dispense solutions into one 24-well Linbro plate (Oldfield et al. system14) due to dispensing times being dependent on solution viscosities, or 2.2 hr to set up one 96-well crystallization tray (Soriano and Fontecilla-Camps ASTEC system15). In addition to vapor diffusion, Chayen et al. proposed using the microbatch under oil method in an automated fashion.16
This chapter discusses about robotics for automated crystal formation and analysis. The choice of crystallization and imaging systems depends on user needs and requirements. Stand-alone workstations will be most appropriate for laboratory efforts or for systems that require added flexibility. Integrated robotic systems will be necessary for more HT industrial needs but with a concurrent limitation on system flexibility and cost. Large-scale structure determination programs, such as the NIH-sponsored structural genomics consortia and other commercial ventures, are relying on the parallel processing of a large number of protein samples that can be completed with this newly developed automation. This ensures that the three-dimensional structure determination can be completed as rapidly as possible for the largest number of targets. Moreover, as with other scientific equipment, the systems described in this chapter will be improved further and the costs can be reduced. Finally, this chapter concludes that small-scale structure determination projects clearly benefit from using these or next-generation robotic workstations, not only for the reliability, lowered cost, and reproducibility that such systems provide, but also for their use in enabling larger explorations of structural space.
Microbatch crystallization under oil - a new technique allowing many small-volume crystallization trials
1992, Journal of Crystal Growth
An approach to rapid protein crystallization using very small samples is described. A computer controlled microdispenser is used to make crystallization samples as microbatch droplets under oil. Samples of 1–2 μl are dispensed ready-mixed and with good precision. The samples are protected from evaporation, contamination and physical shock by the oil. When favourable conditions for crystallization have been found using one mode of the system, the size and quantity of crystals are optimized by a second program which generates a set of conditions throughout the area of interest. Crystals of diffraction size and quality have been grown in 1 μl drops.
Methods of analysis of protein crystal images
1991, Journal of Crystal Growth
Several protein crystallization techniques, including the vapor diffusion method, lend themselves well to automation techniques. Up until the present time, automation techniques have been restricted to setting up crystallization experiments, and procedures to monitor and analyze the experiments have not been developed. These procedures require additional hardware for video monitoring of crystallization chambers and automatic recognition of protein crystals. An automated image acquisition and analysis system makes use of both image processing routines and pattern recognition procedures. In order to design and implement such a system, we are presently developing algorithms which can recognize and locate protein crystals in video images of crystallization droplets. Images of crystallization experiments are acquired and digitized, and analyses of the droplet images are conducted on the microcomputer which also acts as a host in our laboratory robotics system. We describe here our current progress in designing the image analysis system, including the development of appropriate pattern recognition methods. In addition, the usefulness of various pattern recognition schemes for monitoring the progress of crystallization is explored.
A method for programmable control of reservoir concentrations for protein crystal growth
1991, Journal of Crystal Growth
Present protein crystallization methods such as vapor diffusion, dialysis, and liquid dialysis allow little or no control of the equilibration rates. Algorithms and hardware have been developed and tested at UAB which provide manual, interactive, or preprogrammed control of equilibration rates by controlling reservoir concentrations. A variable voltage DC power supply is used to control the flow of a peristalic pump into a microdialysis or vapor diffusion crystal growth chamber. A variety of equilibration curves were constructed initially. The apparatus was then configured to produce the desired equilibration curves experimentally by varying the voltage input to the peristaltic pump. This apparatus can be used to optimize protein crystal growth via dialysis or vapor diffusion techniques.
Lessons from high-throughput protein crystallization screening: 10 years of practical experience
2011, Expert Opinion on Drug Discovery
Macromolecular Crystallization and Crystal Perfection
2010, Macromolecular Crystallization and Crystal Perfection

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Minimal intervention robotic protein crystallization

Abstract

J. Biol. Chem.

J. Crystal Growth

Preparation and Analysis of Protein Crystals

Protein Crystallography