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Automation of protein crystallization trials: use of a robot to deliver reagents to a novel multi-chamber vapor diffusion plate.

We report here the automation of search procedures to rapidly screen a large number of reagents and incubation conditions that lead to the formation of protein crystals. The system consists of a Biomek 1000 Automated Laboratory Workstation from Beckman Instruments under the control of a custom user-interface program developed by Cryschem. A plate composed of twenty-four vapor diffusion chambers, each with its own reservoir well and protein drop holder was designed by Cryschem to fit the Biomek table. The Cryschem software manages a large data base of incubation conditions and generates instructions for the workstation to dispense the protein, buffers, detergents, cofactors and other reagents used to promote the formation of protein crystals. The plate is manually positioned on the Biomek table and then under program control additions are automatically made to each chamber as follows: Precipitating solution is added to each reservoir well and the protein solution along with the precipitating solution and various other reagents are added to each drop holder. The plate is removed from the table and a mylar tape is applied to simultaneously seal all the chambers. The plates are placed at a controlled temperature and periodically examined for crystal formation.

Biotechnology↗

Stabilization of ascorbic acid in human plasma, and its liquid-chromatographic measurement.

We describe two independent HPLC procedures for the rapid, accurate analysis for ascorbic acid in human plasma. No sample extraction or phase separation is required. We also describe a procedure for preparing a human plasma reference material for use in clinical laboratory analysis for ascorbic acid. The ascorbic acid in plasma can be determined in 15 min, with as little as 50 microL of sample. Analytical recoveries are near 100% with direct injection of deproteinized plasma. Extensive stability data under several conditions, with dithiothreitol as a preservative (antioxidant), indicate that ascorbic acid remains stable in stored plasma for as long as 57 weeks. CVs for round-robin analysis of 11 normal human blood samples by two independent methods were between 0.1% and 5.3%. These clinical samples appear to be stable for at least 50 days under the described conditions of stabilization and sample treatment. Finally, because ascorbic acid prepared by the described procedures is stable at room temperature for at least 18 h, these methods can be readily adapted to clinical laboratory automation at room temperature.

Ascorbic Acid↗

The OPTIMIZOR: a graphic pulse analyzer to monitor electro cell fusion and electroporation experiments.

BTX has developed an instrument, the OPTIMIZOR, which allows the accurate monitoring and adjusting of electrical parameters to aid in electro cell fusion and electroporation experiments. The OPTIMIZOR can be used with the family of BTX generators and chambers, as well as home-made or other commercial systems with the appropriate adaptors. The microprocessor-based design allows great flexibility in further expansion of functions and possible mating to a fully-automated laboratory system with robotic features.

Animals↗

Application of robotics to radiopharmaceutical preparation: controlled synthesis of fluorine-18 16 alpha-fluoroestradiol-17 beta.

A commercially available robot system, the Zymark Zymate Laboratory Automation System, has been utilized for the preparation of a positron-emitting radiopharmaceutical, 16 alpha-[18F] fluoroestradiol-17 beta. This radiopharmaceutical is prepared in a three-step synthesis (preparation of [18F]-tetrabutylammonium fluoride, SN2 displacement of a triflate, and ketone reduction) and is purified by high performance liquid chromatography (HPLC). All steps in the synthesis and HPLC purification are controlled by the robot system with no manual intervention. This represents a new approach to the complete automation of radiopharmaceutical production.

Automation↗

Collaborative Behavioral Teratology Study: programmed data entry and automated test systems.

The automated laboratory systems described in this paper were developed for use at the National Center for Toxicological Research (NCTR) and each of five collaborating laboratories. These laboratories participated in a study designed to evaluate the intra- and interlaboratory reliability and sensitivity of several behavioral test methods used in developmental toxicity studies. This paper describes two microcomputer systems. System I was designed to control stimulus delivery to and record behavioral responses from rats during a visual discrimination learning task and to accept data from physical landmark, negative geotaxis, and olfactory discrimination evaluations. The microprocessor system was an S-100 bus, Z-80 processor with 8K PROM, 16K RAM, nine 8-bit parallel I/O ports, and one serial I/O port. For discrimination learning, the animal testing apparatus consisted of six operant chambers, equipped with two stimulus lights, two nose-poke apertures each containing a phototransmitter and receiver, an electromechanical liquid dispenser unit, and a reward bowl also containing a photodiode. Each chamber was interfaced with the microprocessor at a parallel I/O port which permitted the delivery of response- or time-contingent control signals for the lights and liquid dispenser units. System II controlled stimulus delivery and recorded behavioral responses from rats during an auditory startle habituation task, and also recorded rodent activity levels in figure 8 mazes for periods of 1, 4, or 23 hours. The microprocessor system was an S-100 bus, Z-80 processor with 8K PROM, 16K RAM, nine parallel I/O ports, an analog to digital (A/D) I/O board, and one serial I/O port.(ABSTRACT TRUNCATED AT 250 WORDS)

Acoustic Stimulation↗

Accuracy of four methods of home blood glucose monitoring in hemodialysis patients.

The accuracy of four techniques of home blood glucose monitoring in hemodialysis patients was compared. Arterial-venous blood samples were collected from 23 patients undergoing hemodialysis and analyzed for serum glucose concentrations using an automated laboratory method. Capillary-blood glucose concentrations were also determined in each patient by visual readings of the Chemstrip bG and Dextrostix reagent strips and by reflectometer readings of the StatTek Glucose and Dextrostix reagent strips. Readings of glucose concentration obtained by each of the four techniques were compared with laboratory-determined values, and the true and absolute deviations in these values for each technique were recorded. The effect of patient hematocrit on deviations in glucose concentration readings was evaluated by analysis of variance. The mean glucose concentration readings obtained using the visual and reflectometer (Dextrometer) readings of Dextrostix were substantially less accurate than the other two techniques when compared with serum glucose concentrations determined by the laboratory. The StatTek technique gave the smallest mean absolute deviation from laboratory-determined values. No correlation was found between the patient's hematocrit and either the true or absolute deviations in glucose concentrations. The Chemstrip bG and StatTek techniques are reasonably accurate methods for home monitoring of blood glucose concentrations in diabetic patients undergoing hemodialysis.

Blood Glucose↗

Constructing a distributed object-oriented system with logical constraints for fluorescence-activated cell sorting.

This paper describes a fully distributed biological-object system that supports FACS (Fluorescence Activated Cell Sorter) instrumentation. The architecture of the system can be applied to any laboratory automation system that involves distributed instrument control and data management. All component processes of FACS (such as instrument control, protocol design, data analysis, and data visualization), which may run on different machines, are modeled as cooperatively-working "agents." Communication among agents is performed through shared-objects by triggered methods. This shared-object metaphor encapsulates the details of network programming. The system facilitates the annotation of classes with first-order formulae that express logical constraints on objects; these constraints are automatically maintained upon updates. Also, the shared-object communication and polymorphic triggered methods are exploited to produce a homogeneous interface for instrument control.

Automation↗

Networks for electronic radiology.

Next-generation health care systems must successfully integrate hospital information, laboratory automation, and medical-image management to provide a seamless view of the complete medical record to the primary care physician across a distributed medical enterprise. Broadband networks are an important component of the systems required to achieve this goal. This article provides a brief tutorial on relevant network principles and products, a perspective on the evolution of the field, and a simple view of the network requirements posed by electronic radiology.

Computer Communication Networks↗

Tissue impedance as a function of temperature and time.

Tissue impedance dependence on temperature has been measured for six tissue types. The information was gathered using an automated laboratory under computer control. This information is needed to be able to input these values into a computer model that predicts temperature distribution produced by delivery of radio frequency energy. Due to the thermal dose of time and temperature, tissue properties change and no published data are available that document this. Since there are no theoretical predictions, empirical data were measured to supply this information. Using an aluminum cylindrical cavity of volume 1.69 cm3, muscle, liver, brain, and fat tissue impedance were measured at 500 kHz over a range of temperatures. All tests began at room temperature where baseline measurements were made. The cylinder was then placed in a constant temperature water bath at between 30 and 90 degrees C. The tests were run for a period of 10 to 30 minutes. Temperature homogeneity was carefully studied throughout the volume of the cylinder. It was found that thermal equilibrium occurred within four minutes. Special care was taken with the tissue sample in regard to optimize moisture and freshness, and minimize fat content. Grain orientation was also taken into consideration depending on the test. For all tissue types, resistivity decreased initially as the sample temperature equilibrated with the bath temperature. For temperatures less than 75 degrees C, resistivity values remained approximately constant over time.

Adipose Tissue↗

[Sampling and quality assurance].

For the efficient operation of clinical laboratories, automated biochemical analyzers have been improved. Large rapidly analyzing and multichannel analyzers, and small or mid-sized multiple component analyzers are now commercially available and widely use. Such apparatus might require only a small amount of specimen and reagent to assay one analyte. Performance has been increased annually. In this report, we discuss whether this performance, sampling of specimen, sampling of reagent, and mixing effect of reaction mixture or diluted specimen affect the accuracy and precision of measurements.

Autoanalysis↗

Automated external defibrillation: laboratory evaluation.

Twelve samples of ventricular fibrillation were fed into nine automated external defibrillator-pacemakers ( AEDP , "Heart Aid") of recent design. All the devices recognised and defibrillated ventricular fibrillation in seven of the samples within 30 sec. None of the devices reacted to two of the samples; in the remaining three there was inter-device variation ranging from an appropriate response to no response, as well as inappropriate pacing or delay in recognition and treatment. Poor recognition of some ventricular fibrillation waveforms with considerable inter-device variation limits the usefulness of this model. A new prototype responded more consistently and future models may be of value in community resuscitation. The difficulty of evaluating the diagnostic capability of AEDP devices in clinical use makes comprehensive laboratory testing essential prior to release.

Electric Countershock↗