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.     

Developing an Automated Enzyme Immunoassay for High-Throughput Screening of Bovine Serum Antibodies to Neospora Caninum

An enzyme-linked immunosorbent assay (ELISA) for Neospora caninum antibodies was automated with a robotic workstation, the Beckman Coulter Biomek 2000, to screen 200 bovine sera. Comparing these results with manually run ELISA data, a 95.92% agreement (K = 0.9592) between the two assays was obtained. The automated assay was specific and sensitive with excellent positive and negative predictive values. The results were repeatable and reproducible. The automation flexibility was high and the operation complexity was minimal. High-throughput screening (HTS) for bovine antibodies to Neospora caninum was achieved. The assay was developed according to the internationally recognized ISO17025 standard requirements.     

Cellular Telephone-Aided Real-Time Reporting System for Emergency Laboratory Test Results

The authors established a system for real-time reporting of clinical laboratory test results utilizing the mailing function of a cellular telephone service. The reporting system is composed of systems for sample transportation and automatic analysis, verification of test results, and reporting of test results. The introduction of the system has completely eliminated time delays at all stages of the laboratory testing process from start of testing to reporting of the test result. This has enabled doctors to receive test results in a timely manner wherever they are and to respond promptly to the patient condition.     

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.     

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.     

Standardizing Laboratory Data Interchange in Clinical Trials

The Clinical Data Interchange Standards Consortium has developed a Laboratory Model for laboratory data that is generated during the conduct of clinical trials. The Laboratory Model is the first step in proposing standards for the interchange of clinical trial laboratory data. Standards will decrease the time and resources required by stakeholders in the pharmaceutical development process (pharmaceutical companies, biotechnology companies, contract research organizations and laboratories). Standardization will therefore contain costs as well as improve data quality.     

Project Control for Laboratory Automation Outsourced to Consultants A 10-Step Process to Optimize the Effectiveness of Custom Information Technology Development

This article outlines a 10-step process for business leaders in the pharmaceutical, biotech, health sciences, and clinical fields, who hire information technology (IT) applications consultants to design, develop, implement, and integrate custom laboratory automation systems. The goal of this model is to identify steps to dramatically improve the effectiveness of the consultant and to reduce implementation risk factors. The probability of project success can be increased significantly when the basics of IT systems development is understood by those internal to the organization and management guides the consultant to the finish line using a defined project control process.     

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.     

A Flexible and Robust Peer-to-Peer Architecture with XML-Based Open Communication for Laboratory Automation

Traditional lab automation systems are highly centralized: dispatch and coordination of activities are mediated by a system controller, usually via a single, monolithic control procedure. This approach, while conceptually simple, makes changes to the system difficult; adding or removing instruments and functionality can be a daunting task. In addition, most automated systems are tied to particular development languages and protocols, making operation in heterogeneous environments (i.e., the real world) problematic, since instrument software comes in many different implementations.We present a peer-to-peer architecture for lab automation, using an XML-based communication protocol. The architecture consists of peer instrument servers, an XML communication layer, and an open control center. Each instrument peer can control, be controlled by, and communicate information to other instrument peers to fulfill the automation task. Our protocol is based on XML-RPC, a lightweight communication standard built atop HTTP.This provides an open and flexible means of peer-to-peer interfacing. The control center serves as a convenient, Web-based interface to manage the instruments. The automated procedure can be distributed across all available instrument peers (each instrument assigned a set of responsibilities); the controller implements a limited set of high-level instructions. The software components included in our prototype system are implemented in various programming languages, including Java, C/C++, Visual Basic, and LabVIEW. Our approach facilitates rapid development of laboratory automation systems.     

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.     

Incorporating Automation in Undergraduate Laboratories

Automated techniques and philosophies, which have become increasingly important in modern laboratories, are not traditionally covered in undergraduate chemistry curricula. To this end, we have been incorporating automated sample preparation methods using standard robotic workstations in our undergraduate analytical chemistry laboratory. Using a Benchmate™ II Workstation, an automated method has been developed and implemented for the solid phase extraction of capsaicins from commercial hot pepper sauces prior to liquid chromatographic analysis. This paper reports on pedagogical aspects of incorporating automation into the undergraduate curriculum as well as results obtained for manual and automated extractions conducted by students.     

Converting a Protease Assay to a Caliper® Format LabChip System

The push for higher throughput screening coupled with the desire to use smaller volumes of material has sparked the development of new technologies. Caliper Technologies, Corp. (Mountain View, CA) has designed a microfluidics chip with unique properties yet to be fully exploited. The translation from a traditional plate-based assay to a microfluidic chip format has provided insights into assay development, screening data requirements, and the technology itself. Running a screen with this new technology presented challenges in throughput, signal acquisition from slow-conversion enzymes, the provision for a negative control, the translation of a time series into a single data point per compound, reagent adhesion in the channels, and fluid property mismatches. Overcoming these obstacles has resulted in a simple, robust system with significant savings in reagent use. Measures to improve throughput and generalize the system will be discussed.     

Automated High-Throughput mRNA Selection From Eukaryotic Total RNA

We describe a new technology (patent pending) for high-throughput selection of poly(A)⁺ RNA from total RNA. A novel binding solution is used to ensure the efficient and specific binding of mRNA to oligo(dT) magnetic beads with high stringency, virtually eliminating the non-specific binding of ribosomal RNA (rRNA) either to oligo(dT) beads or to the poly(A)⁺ RNA bound to the beads. As quantified by real-time RT-PCR, more than 99% of the rRNA is removed in a single round selection and mRNAs are fully recovered for both highly-expressed (GAPDH) and poorly-expressed (DDPK) genes from a few μg total RNA. The protocol is adaptable to any generic robotic workstation and takes ˜30 minutes to process 96 samples.     

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.     

Controlling a Twister II with VBA and the Zymark ZyRobot ICP

A Visual Basic for Applications program was developed for direct control of a Twister II robot using the Zymark ZyRobot_ICP. Direct control enables automated check-out of the robot, and automated measurement of the plate stacks prior to a robot run. Normal operation of the robot program is performed with just two buttons.     

ASTM E1989-98 - The New Instrument Control Standard

The new ASTM E1989-98 Laboratory Equipment Control Interface Specification (LECIS) is a robust standard definition of equipment behavior while under remote control. The goal of the standardization effort is to facilitate “plug-and-play” integration of laboratory automation with standard hardware behavior and software interfaces. The LECIS standardizes laboratory equipment behavior and a message passing scheme between the controller and equipment that synchronizes this behavior. Commercial adoption of this new standard is well under way.     

Assay on FAQ''s on Total Lab Automation

The purpose of the introduction of an automated laboratory system in one format or another is reducing the costs of the laboratory by cutting staff. Also speeding up the performance often is a goal. At the same time it is planned to increase the quality of the work done by using automated labelling and reading of documents. The use of vacuum tubes and the handling of these by the apparatus are raising the safety of the labwork.

In this paper many aspects of the daily routines when using a TLA system are discussed. The aspects are collected during the discussions before and after the implementation of the Roche CLAS system in our laboratory. Some consequences were foreseen, and others came unexpectedly. But still the subjects form a point of discussion around the TLA systems. The most important message to be picked up is that one really has to consider all possible problems that could arise from the implementation of TLA.     

Flexible and Scalable Automation Solutions For Scoring Single Nucleotide Polymorphisms Using Rolling Circle Amplification

A single nucleotide polymorphism (SNP) scoring assay that uses ligation-dependent Rolling Circle Amplification (RCA)† was transferred to a series of automated protocols addressing a range of throughput levels. The systems utilised various automation modules consisting of custom-made and offthe-shelf devices. Several system parameters were evaluated to ensure assay integrity and homogeneity. These included reagent carry over, liquid evaporation rates, thermal regulation of reactions and fluorescence reading capabilities.Data analysis software was developed in order to rapidly allocate SNP calls from data generated by the automated system. A modified fuzzy c -means clustering algorithm was employed to separate data points into groups associated with specific genotypes. Data were then presented graphically and within a summary table, which allowed easy and rapid organization and interpretation of data.