Journal of Laboratory Automation

Double-stranded DNA binding sites that bound with high affinity to the nuclear factor kappa B (NFκB) p50 homodimer were selected using a Tecan Genesis workstation. The adaptation of the Tecan to automated selection required the integration of multiple devices and modifications to standard selection protocols, and resulted in a significant increase in throughput. The sequences obtained by automated selection strongly correlated with the well-known family of natural NFκB double-stranded DNA binding sites and with previous manual selection experiments. In addition, the selection experiments better defined the contributions of residues outside of the well-known, decameric core binding site for NFκB.

Automation will be a necessary step for the success of molecular computing. Here we have applied microfluidic technology to DNA computers, with a sample application towards solving Boolean logic problems.

This article reports on a novel dispensing system for the massive parallel delivery of liquid volumes in the range of 50 nL. Due to the similarity of the device to conventional micro-well plates used for the storage of liquids, the device has been termed “dispensing well plate” (DWP). In contrast to other known micro dispensers, the DWP can consist of up to 1,536 dispensing units in parallel, all of which hold different reagents. The dispensing units can be arranged very closely at the pitch of conventional micro-well plates (2.25 mm or 4.5 mm). Driven by pneumatic actuation, a fixed volume of different liquids can be dispensed simultaneously and contact-free into micro-well plates or onto flat substrates. Because of this, the liquid handling in many chemical, biochemical, and pharmaceutical applications—especially within high-throughput screening (HTS)—can be sped up by a factor of 10 to 100. In our article, the basic operation principle of the device is presented, and experimental evidence of its extraordinary performance is given: a reproducibility of 2% to 5% and a homogeneity within individual droplet arrays of 1% to 2% has been measured, as well as viscosity independent performance for liquids in the range from 1 to 5 mPas. The applicability of the DWP technology within HTS is demonstrated by performing a miniaturized kinase assay at 1 μL assay volume in a 1536-well plate format.

This paper reports on a simple, disposable non-contact dispenser for the nano- and microliter range. In contrast to other known dispensers manufactured by silicon micromachining^1-4 the new device simply consists of an elastic polymer tube with a circular cross section. Actuation is done by a piezostack driven piston, squeezing the tube at a defined position near the open end by a significant fraction of the cross section. In contrast to drop-on-demand devices based on an acoustic actuation principle,⁵ the squeezing of the tube leads to a significant mechanical displacement of the liquid. Our experiments tested a large number of media in the viscosity range from 1 to 27 mPas. Some of our experiments tested up to approximately 2,000 mPas. Frequency characteristics showed an independent dosage volume for water up to a frequency of 15 Hz for tubes with an inner diameter of approximately 200 um. Standard deviation within 1,000 shots resulted in an excellent CV (standard deviation/dosage volume) of less than 2% of the dosage volume. Using tubes with an inner diameter of approximately 1,000 um and a print frequency of 340 Hz, a flow rate of less than or equal to 143 μL/s could be reached. Beyond the possibility to dispense pure liquids, emulsion paints with particles that have a diameter of approximately 40 μm have also been printed successfully.

In December 2001, the McMaster HTS Lab officially opened its doors. Since that time, we have made significant strides in demonstrating that university-based, high-throughput screening (HTS) is a viable proposition for academic scientists seeking to discover novel small molecule probes of biological function. Although the lab has been running screens for just over two years, the process of designing, building and maintaining the lab has been on-going for more than four years. As high-throughput screening technology moves from the industrial sector to the academic and small biotech sectors, strategies for setting up a successful, highly flexible HTS lab on a limited budget becomes very important. In the current communication, we outline some of the considerations in setting up the lab and some of our experiences to date with screening and automation in academe.

The present study evaluates the application of Nanostream Inc.'s Veloce micro-parallel liquid chromatography (μPLC) system, a high-throughput microfluidic liquid chromatography (LC) system, for the analysis of peptides and proteins. Using the Veloce system, as well as traditional high-performance liquid chromatography (HPLC) systems, we performed separations and analyses of (1) a mixture of peptide standards, (2) crude and semipure synthetic peptides, and (3) peptide components of a protein digested with trypsin. In all these studies, the Veloce system was evaluated by comparison to a traditional HPLC system for the number of peaks, their resolution, retention time, shape, and width. When similar columns were used, the performance of the Veloce system was comparable to that of traditional HPLC systems. In general, the Veloce system offers several advantages over traditional HPLC systems, such as the ability to analyze samples in a high-throughput manner with significantly lower consumption of samples and solvents.

A critical need exists for the development of next-generation genomic analysis instrumentation capable of offering significantly higher throughput at a lower cost than current technology. In this paper, we explore the potential of natural convection-based systems to address these issues by providing a thermocycling hardware platform capable of performing rapid polymerase chain reaction (PCR) amplification of DNA. These systems can be arrayed in a multi-well format that is simple to operate, is suitable for integration with high-throughput automated liquid handling systems, and can be easily and inexpensively mass-produced.

A cell physiometry system is described for characterizing and separating cells, and performing cell-based assays, using dielectrophoresis (DEP). Cells, or mixtures of cells, are suspended in a chamber containing an array of microelectrodes located on the chamber's bottom surface. A sequence of radio frequency signals is automatically applied to the microelectrodes, and images of the DEP-induced motions of the cells are captured and analyzed to determine a characteristic parameter known as the DEP cross-over frequency. Once a cell population has been characterized in this way, the same apparatus can be used to selectively isolate cells for additional biochemical, physical, or genetic analysis. Biochemical labels or bioengineered tags such as fluorescent markers or antibody-coated beads are not required. Cell separations are achieved by “electronically tuning” into the different cell types by superimposing different signal frequencies onto the microelectrodes, using both stationary and traveling wave DEP signals. Examples of separating different types of blood cell are given, together with descriptions of cell-based bioassays that monitor physiological changes that accompany trans-membrane signaling events, apoptosis, and the aging of cell cultures.

A novel curriculum in laboratory automation and high-throughput technologies has been developed at the Keck Graduate Institute (KGI) over the past five years as part of the professional masters degree program in applied life sciences. The goal of this curriculum has been four-fold: (1) motivate study by describing the need for automation through several example problems in combinatorial biological discovery, (2) provide elements of fundamental engineering science that are required for the development of the technologies and tools that enable automation, (3) provide opportunities for the students to see and use state-of-the-art instruments, learn about existing industry standards, and to visit integrated laboratories that perform high-throughput research, and (4) introduce scientific discoveries and new technologies that could have future impact on laboratory automation and discuss current trends, and project future trends in this field.