HTS Automation Study: Results from a 2001 Survey of the Current vs. Desired State of HTS Automation

A survey of directors of screening organizations was conducted in 2001 to evaluate their perceptions of the current vs. desired state of high-throughput screening (HTS) automation. The survey encompassed attributes such as automation flexibility, throughput and operation. These and other automation attributes were ranked based on importance to the respondent and/or the limitations these attributes imposed on the screening organization.     

The Second Generation of the Laboratory Information System and the Laboratory Automation System in Osaka City University Hospital

The first total clinical laboratory system (TCLA) in the Osaka City University Hospital was introduced in 1993. After six years of operation, it was recently replaced by a new system. The goals of this replacement were as follows: 1. Improve the analytical performance. 2. Make the system operate more efficiently. 3. Improve the quality of laboratory analyses. We successfully reduced the labor required to operate the laboratory; made the laboratory reports quicker; reduced the number of retraction and revalidation of the results; and minimized the system downtime.     

Engineering An Experimental Platform For High Throughput Reactivity Screening

The genomics revolution coupled to advances in computational power, informatics and robotics is driving drug discovery programs to produce drug candidates faster. This need has resulted in advances in high throughput methods for performing organic chemistry such as combinatorial and parallel synthesis. Yet there has not been a corollary advance in the ability to collect quantitative information on reactions that can be used to produce these drug candidates. This lack of an efficient and robust analytical method has resulted in a significant chemistry bottleneck. This work outlines a set of methods that helps address this chemistry bottleneck by using analytical constructs to detect and quantify reaction outcomes. To accomplish this, an integrated experimentalcheminformatics platform has been developed which couples an experimental design system, automated high throughput parallel and combinatorial synthesis methodology, sample processing, quantitative mass spectroscopy and automated data analysis. This platform is being used to optimize single reactions and the syntheses of whole libraries of compounds, and to generate large databases on specific reaction classes.     

Selective Ion Extraction for High Throughput Screening with a Microfluidic Enzyme Assay

This proof-of-concept paper describes the application of selective ion extraction to an assay of protein kinase A on a microfluidic chip platform. Selective ion extraction is a flux balance technique, where a combination of independent pressure control and voltage are used to selectively extract one ion from a mixture. The assay product is completely separated and diverted into a separate channel from the waste stream containing the unconverted substrate and enzyme. By detecting only product, background noise generated by the substrate is removed which increases the signal to noise ratio and assay sensitivity. This technique is intended for adapting kinase or protease assays with low conversion rates to an on-chip reaction format for HTS screening.     

Transferring Industrial Automation Technology to the Laboratory

Over the past few years, there has been much discussion about transferring industrial technology to laboratories. While it is easy to look at the superficial similarities, it is more important to examine the different requirements of different industries. In this way, it is possible to identify the technologies and techniques that can be successfully transferred to the laboratory to improve performance.This paper takes three very different industries - the bakery, High Throughput Screening (HTS) and mobile phone assembly and examines their different requirements. These industries have been selected from among the many sectors where the RTS Group operates - thus allowing real data from a number of situations to be used.One of the most important areas in automation design is the relationship between flexibility and throughput. This paper focuses on this relationship and its influence over machine configuration when comparing the requirements of the different industries.     

Optimizing Reaction Conditions of the NanoOrange® Protein Quantitation Method for Use With Microplate-based Automation

Because of the rapidly expanding need for higher sample throughput in drug discovery, automation of corresponding biochemical analyses is desirable. In particular, automation of protein quantitation is crucial since its results are used extensively. Recently, a single-reagent fluorescent protein quantitation method (NanoOrange®) with attractive performance attributes has become available. While it can potentially be automated with liquid handling workstations, several of this method's reaction parameters need to be optimized.

We studied the time and a temperature dependence of the NanoOrange protein quantitation reaction in ninety-six well black microplates using either a temperature-regulated hot block or a microwave oven as heat sources. Fluorescence of the NanoOrange reaction was quantified with a multimode microplate spectral scanner.

Time-dependent heating profiles of filled microplates placed on hot blocks at fixed temperatures (45, 55, 65, 75, and 95°C) revealed temperature differences of 4–7°C cooler for the outside wells compared to the inner wells, however the maximum well temperature did not exceed 65°C. Similar time-temperature studies of microwave-heated microplates revealed an equilibrium temperature of 45–49°C that was 10–16°C lower than microplates that were block heated.

The bovine serum albumin (BSA): NanoOrange standard curves created using a hot block increased in slope from 45°C to 55°C, but then remained constant from 65 to 85°C.

Fluorescence of the BSA: NanoOrange standard curve created using a microwave oven was about half the magnitude of the hot block-derived curves, possible reflecting a lower energy transfer rate of the microwave oven. We conclude that the NanoOrange protein quantitation method can be automated if a microplatecompatible hot block with a 65-85°C surface can heat the microplate for minimum of 15 min prior to quantifying the reaction's fluorescence.     

Automation for High-throughput Identification and Picking of GFP Expressing Colonies

Green fluorescent protein (GFP) has many applications as a marker in living cells, and has become widely used as a reporter gene in microbial, plant and animal cells. Screening microbial colonies for GFP expression enables various types of assays (e.g. for secretory mutations). However, this is laborious, non-quantitative and potentially hazardous to the operator (due to UV illumination) when performed manually. In order to address this the GloPix robot was developed. The imaging system discriminates between colonies based on the level of fluorescence activity and the picking function automatically transfers cells to microplate wells. Measuring fluorescent activity allows quantitation of fluorescent tag concentration/expression.     

Automation of the Pre-Analytical Phase: A Performance Evaluation of Alternative Scenarios

This paper is about the automation of the pre-analytical phase in a biochemical laboratory, which performs more than 3.7 million tests per year. The paper presents how an investment in the laboratory can be evaluated considering both economic criteria, future performance and service quality. The study comprises a workflow analysis of biological materials and information, the corresponding data collection and the use of simulation for performance evaluation. Alternative scenarios have been considered in terms of personnel, pre-analytical devices and management policies. The final scenario has been chosen according to economic criteria among a set of feasible scenarios, able to satisfy constraints.     

An Introduction to Using XML for the Management of Laboratory Data

Extensible Markup Language (XML) and the XML Path Language (XPath) are introduced with software examples demonstrating how one can use them to write laboratory data management programs. Topics explored include XML document creation, manipulation, and searching. Programming examples make use of the Microsoft® XML Parser library and the Visual Basic programming language. The problem of managing microplate screening data is used as an illustration. Source code for all examples can be downloaded from http://www.labprogrammer.net.     

Automated High Throughput Screening of Fluorescent Imaging Plate Reader (FLIPR) Assays Using Assay Platform™ Integrated with Activity Base for ‘on the fly’ Compound Reconfirmation

Advances in the field of automation have meant hitherto complex manual cell-based assays can now be automated. These improvements have brought significant enhancements in throughput, data fidelity and consistency, and allowed a reallocation of constrained resources.Building upon these improvements, we have linked our automated cell-based screening system, Assay Platform™, to Activity Base (IDBS), a software package designed to automate the analysis of HTS data. Customisation of this package has resulted in software that can identify ‘active’ compounds and re-pick them ‘on the fly’ from the original compound plates for triplicate re-testing without operator intervention.Based on an operator initially defining ‘normal’ parameters for assay activity in Activity Base, combined with an automated quality control software module that checks data fidelity, wells containing ‘active’ compounds can be re-picked and re-tested at the end of an automated screening run. Automating cell-based assays has significantly improved productivity, and, with the synergism of Activity Base, has given us greater power to complete each screening run and report ‘active’ compounds to Chemistry more rapidly. This article presents our approach to the automation of cell-based Fluorescent Imaging Plate Reader (FLIPR) screening together with automated active re-test confirmation using Activity Base.     

A simulation model for eliciting scheduling knowledge: an application to the precast manufacturing process

This paper discusses the process of eliciting scheduling knowledge from a simulation model and the development of a dynamic modelling approach to the scheduling process in the precast concrete industry. Due to the problems associated with eliciting scheduling knowledge from an ‘expert’ in the precast industry or perhaps in most of the manufacturing industries, simulation is used to complement human knowledge in this paper. Such knowledge will be used for online support to advise production schedulers and for further development of the simulation model by incorporating the knowledge in the model and making it more dynamic. The paper suggests that dynamic selection of scheduling rules during real-time operation has been recognised as a promising approach to the scheduling process in the precast industry. For this strategy to work effectively, sufficient knowledge is required to enable the model to predict the most effective scheduling rule to meet current factory status. The paper concludes that if the knowledge rules are used effectively, they could be a considerable managerial tool for exploring and improving managerial practices. Recommendations have been made regarding the development of a more realistic and practical scheduling system.     

Remarks on two fixed-point theorems involving the sum and the product of two operators

In this article, we focus our attention on two fixed-point theorems of Krasnoselskii [1] and Dhage [2]. It is shown that some of the hypotheses of these fixed-point theorems are redundent. Our claim is also illustrated with an example.     

Detection of Cellular Parameters Using a Microfluidic Chip-Based System

Lab-on-a-chip technology achieves a reduction of sample and reagent volume and automates complex laboratory processes. Here, we present the implementation of cell assays on a microfluidic platform using disposable microfluidic chips. The applications are based on the controlled movement of cells by pressure-driven flow inside networks of microfluidic channels. Cells are hydrodynamically focused and pass the fluorescence detector in single file. Initial applications are the determination of protein expression and apoptosis parameters. The microfluidic system allows unattended measurement of six samples per chip. Results obtained with the microfluidic chips showed good correlation with data obtained using a standard flow cytometer.     

Analysing the hypoglycaemic counter-regulation: a clinically relevant phenomenon?

This paper describes and analysis of the temporal relation between episodes of low blood glucose (hypoglycaemia) and counter-regulations, i.e., episodes of elevated blood glucose (hyperglycaemia), in patients with insulin dependent diabetes. The relation was assessed by statistical methods based on a metabolic computer model of the human glucose metabolism. The study material was standard collected clinical data on meals, insulin injections, and measured blood glucose from hospitalised patients. We have found that a typical hypoglycaemic counter-regulation begins 6–8 h after the hypoglycaemia, that it lasts 16–18 h, giving a total duration of 24 h, and that it elevates the blood glucose by 4–10 mmol/l. The phenomenon was demonstrated in the data from more than half of the patients with hypoglycaemic episodes.     

On the preservation of observability under sampling

In this paper, we investigate the problem of the preservation of observability under sampling.     

Object-Z: A specification language advocated for the description of standards

The importance of formalising the specification of standards has been recognised for a number of years. This paper advocates the use of the formal specification language Object-Z in the definition of standards. Object-Z is an extension to the Z language specifically to facilitate specification in an object-oriented style. First, the syntax and semantics of Object-Z are described informally. Then the use of Object-Z in formalising standards is demonstrated by presenting a case study based on the ODP Trader. Finally, a formal semantics is introduced that suggests an approach to the standardisation of Object-Z itself. Because standards are typically large complex systems, the extra structuring afforded by the Object-Z class construct and operation expressions enables the various hierarchical relationships and the communication between objects in a system to be succinctly specified.     

Global controllability of linear autonomous systems: A geometric consideration

The global controllability in finite time of a linear autonomous system with restrained controls is investigated. Necessary and sufficient conditions are obtained by an approach based on the consideration of geometric properties of the system.     

Automation of Chemical Reaction Kinetics and Product Distribution Studies in Pharmaceutical Development

An automated instrument was designed and constructed to facilitate the performance of pharmaceutical degradation studies. A brief theoretical background on degradation kinetics is given to rationalize the design of the instrument and representative data are provided to illustrate its successful application. This system was found to be capable of conducting multiple simultaneous isothermal and nonisothermal kinetic studies with user-defined temperature profiles, sampling periods, and data logging.