Laboratory automation is changing the work flow in biochemical and chemical laboratories. In smaller labs, automating the mundane processes improves data quality and reduces labor. During the last year, several vendors have introduced instruments and modules designed to improve work flows. These are described in the new products section below. The upper end of the automation segment is moving ahead with new high-speed, high-resolution confocal scanning instruments, which record and analyze label-free (LF) images of living cells. These scanners are fast—fast enough to enable making kinetic scans at several time points—and produce big data files. Big Data is probably today’s sorest pain point, but the Big Data problem will get worse over the next few years. Circulating tumor cells are showing utility for patient stratification and guidance of therapy. Thus, the leading edge of high content analysis will shift from supporting data-rich research to clinical diagnostics. This requires multiple whole-genome assays per patient. With 1.7 million new cancer cases expected in the U.S.A. by the end of 2012, high sample load will require huge advances in automated PCR assays and informatics.
Lab automation conferences
Laboratory automation provides engineering support for innovative new work flows in research, quality assurance, and diagnostics laboratories. Developers gather at three meetings each winter to communicate the state-of-the-art and contemplate new capabilities.
In 2013, the Meeting of the Society for Laboratory Automation and Screening (SLAS) will switch coasts to Orlando, FL (January 12–16) (www.slas.org). Major program tracks include: Assay Development and Screening, High Throughput Technologies, Micro/Nano Technology, Informatics, and Bioanalytical Techniques. In prior meetings, the technical sessions enjoyed a symbiotic relationship to the exhibition. Poster sessions are usually short but intense with numerous applications reports. Keynote lecturers present views into the future. This year, Prof. Mehmet Toner of Harvard Medical School and Massachusetts General Hospital (Boston, MA) has been invited as a keynote lecturer discussing “Bioengineering and Clinical Applications of the Circulating Tumor Cell Microchip.” Prof. Toner is known for his work in isolating circulating tumor cells (CTCs) with microchip devices. He has shown that a count of CTCs in 7.5 mL of blood is useful in classifying patients and guiding therapy for breast cancer. The chips show particular promise for detection and therapeutic responsiveness of drug candidates. Tumor stem cells are a related topic, since metathesis to nonadjacent tissue may involve transport in blood.
Cells in cancerous tumors are heterogeneous, presenting a range of activity. Analysis of the PCR amplified DNA in single CTCs shows scrambling of the DNA between the genes. For example, genes that produce growth can be separated from their stop signals. Connectivity of the genes is thus very important, hence the interest in single cell isolation and whole genome sequencing. It is probable that one will need to do whole genome analysis of several to many of a patient’s CTCs to characterize the cancer and look for matches with therapeutic opportunities. If DNA sequencing of CTCs is shown to have therapeutic utility, then one can expect a huge increase in automating cell capture, preparation for DNA sequencing, sequencing, and data processing. The latter seems particularly challenging since the files are potentially so large. This will necessitate tremendous increases in Internet capacity and data processing power. On the research side, the availability of data sets of CTC genomes from hundreds of thousands of patients will require significant enhancements in data integration, interrogation, and display. Since the patient population is large, these units will probably be core facilities in regional centers, at least initially.
AACC meeting series: lab automation for clinical laboratories
Automation in clinical labs is the focus of the meeting series organized annually by the American Association for Clinical Chemistry (AACC). The 2012 meeting is scheduled to be held December 6 and 7 in Tampa, FL (www.aacc.org). Topics include: Task-Targeted Automation; Advances in Preanalytics; Role of Continuous QA/QC in Process Optimization; Command Controls/Data Handling; and Automated Non-Chemistry Applications for the Clinical Lab: Microbiology, Coag, and Hematology.
High Content Analysis (HCA) 2013
HCA 2013 is a meeting that deals with very large-scale screening and high content analysis (www.highcontentanalysis.com). High content analysis systems are found in core facilities staffed with about 50 scientists involved in all aspects of screening, from assay development to data storage and retrieval. The scale differentiates these facilities. An experiment may involve 10 million wells and generate a few terabytes of data in a day or two. The agenda of the tenth HCA meeting includes: Compound Screening and In Vitro Toxicology, HCA for Pathway Analysis and Target Validation, Live-Cell and Label Free Imaging, Analysis and Data Management, HCA of Stem Cells, and Phenotypic Drug Discovery. HCA 2013 will be held January 8–12, 2013 at the Fairmont Hotel in San Francisco, CA.
Kinetic analysis and label-free cell imaging
Hot technologies are expected to be kinetic analysis and label-free imaging of single live cells. The latter is important since labels are suspected in perturbing living cells, which reduces data quality and utility. In the early days of HCA, assays were run for a fixed-time endpoint and read. Biological systems show a range of kinetic responses. This information was lost in the early days.
As a field, high content analysis started in about 2001. It rapidly progressed from a collection of modules assembled on a large tabletop to today’s integrated imagers supported by huge data farms. The current state-of-the-art on integration is illustrated in a report from Yokogawa (Tokyo, Japan), Wako Automation (San Diego, CA), and Kalypsys (San Diego, CA), which targets HCA of single cells.
Yokogawa extended the Cell Voyager line to the CV7000 Live Cell Imaging system. This instrument simultaneously collects confocal images in three colors, using a larger scientific complementary metal oxide semiconductor (sCMOS) camera that quadruples image area. This enables rapid imaging of 384-well plates in 4 min with a 50-msec exposure time. sCMOS cameras are unique in their ability to simultaneously offer ultralow noise, rapid frame rates, wide dynamic range, high resolution, and a large field of view. The environment of the stage is also fully controlled for temperature, humidity, and CO2. Photobleaching is reduced by dual Nipkow spinning disk technology, which allows kinetic assay of live cells over several days.
The CV7000 requires a plate handler system to prepare target plates and feed them to the imager. Thus, Wako Automation developed a robot driven by Kalypsys Director™ software. Results from the high-throughput screen (HTS) can be used on-the-fly to select and route plates with interesting wells for further analysis. This saves time and reagents and improves work flow by focusing on interesting results.
Kalypsys Director software uses input from HTS results calculated on-the-fly to automatically write new robotic methods that only send plates with interesting data to a high content imager so that selected wells can be imaged for further analysis. The powerful and unique features of Director software save time, reagents, and disk storage by focusing on collecting better data. Plus, the speed and programming flexibility enable following developments for extended periods.
Promise of semantic technology
Big Data is still a problem for large-scale screening. Integration of large data files is frustrating. Semantic technology (ST) makes this easier, but depends on a domain- specific ontology to help convert relational data files to RDF format. IO Informatics (Berkeley, CA) has a suite of programs that facilitate the transition plus power the interrogation of the updated or combined database. ST is also showing promise in searching natural language documents. Electronic medical records, which combine relational data files with text, are one obvious application target.
Laboratory automation for the masses
During the last year, several vendors have introduced products that should be very useful for automating laboratory work flow. Some will be justified by improving data quality, and others economics. Even in China, where labor is not expensive, labs find that automation improves data quality. It is a case of eliminating humans and our associated variability. The following is a sampling of new products introduced in 2012.
HP (Palo Alto, CA) is widely recognized for inkjet printers, where reagents are precisely measured and transferred without contact to the target (paper). HP and Tecan AG (Männedorf, Switzerland) recognized that this dispensing technology could also be useful in contactless dispensing reagents for assays into multiwell plates: hence the HP D300 Digital Dispenser from Tecan AG. The device is able to dispense volumes ranging from 13 pL to 10 μL with an accuracy of better than 8%. Solvent compatibility includes aqueous and most organics, i.e., dimethyl sulfoxide (DMSO). Dead volume is less than 2 μL.
The D300 is particularly useful for titrations such as IC50s in multiwell plates. Loading the plates is easy and fast. Serial dilutions and associated errors are avoided, since each well is loaded independently. Reagent consumption is reduced dramatically, as are pipet tips (www.tecan.com).
Integra (Zizers, Switzerland) introduced the VIAFLO 96 Pipette for reformatting plates and serial dilutions. The workstation features a control stick that the operator uses to control instrument functions, including tip loading, aspirating, and dispensing. The tip array can contain 96 tips for plate loading and reformatting. This is about seven times faster than a conventional eight-tip strip pipet. For serial dilution, the 96-tip head is replaced with a “strip of 8,” which stirs, aspirates, and dispenses as the head moves to the next row (www.integra-biosciences.com).
Integra introduced a new liquid handler—the VIAFILL Multifunctional Reagent Dispenser—that combines many options in one small unit. With the VIAFILL, the user can change the mode from bulk liquid dispensing to multiwell plates. In Multiwell Pipette Mode, one can aspirate from and dispense to individual wells, as in serial dilutions. In Plate Washer Mode, liquids can be dispensed, agitated, and aspirated as required (www.integra-biosciences.com).
Smart sample concentrator
Sample blowdown is a common protocol that is often poorly executed, especially manually. Fluid Management Systems (FMS, Watertown, MA) introduced the SuperVap™ for automated direct-to-vial sample concentration. This preserves sample integrity and avoids incomplete transfers or the need to add liquid to wash the concentrate to a vial. The SuperVap can handle up to 24 2-mL vials per run. Maximum vial size is 220 mL. Concentration programs are either time- or endpoint-based. Temperature control is with dry heat, thus avoiding the expense and mess of water baths. The human interface is via a clear touchscreen panel or remotely via computer (www.fmsenvironmental.com).
Glygen (Columbia, MD) introduced two new plates that will change the work flow in many labs. The SlitPlate™ adapts Glygen’s SlitTube™ technology to the plate format (www.glygen.com). With the SlitPlate, each well has a very fine slit cut in the plastic bottom of each well. This is so fine that solvents do not leak out, except when spun in a centrifuge. Solids are retained since they are too big to pass through the slit. Thus, the work flow can be simplified by eliminating aspiration and dispense steps. The company also introduced the iPlate™, which has only 1 μg of sorbent in each well. This facilitates processing of very small samples.
Problem-free seals for multiwell plates
Even after more than two decades of use, sealing of multiwell plates is the source of problems. The seals often plug needles, fail at –80 °C, or contaminate the sample with adhesive, to name the most common. The RAPID EPS S is a new plate sealing system from Bio Chromato, Inc. (Fujisawa, Japan) that solves these problems. Plus, it is compatible with the common organic solvents such as DMSO. The seals are available as individual sheets or in rolls for automated plate sealers (www.bicr.co.jp).
From the beginning of the 1980s, microfluidics offered potential savings in time, reagents, and supplies. However, time to implementation was longer than anticipated, at least by the most impatient potential users. Gyros from Covance Laboratories (Chantilly, VA) is one example; however, a recent report from the company illustrates that the promise is being fulfilled. The case in point was an assay for free insulin in human serum. The Gyros method was six times faster and used only 1 μL of sample, which is a small fraction compared to legacy protocols.1
Advances in materials science
Novel applications of advances in materials science should soon change work flows in the laboratory. For instance, a team in the laboratory of Prof. Yang Yang2 at UCLA reports on “Visibly Transparent Polymer Solar Cells Produced by Solution Processing.” Thus, the surface of an instrument case might serve as the source for electrical power to power the analyzer and transmit the results from nearly any location other than coal mines and bank vaults. Initial conversion efficiency was 4%, but the technology is new and likely will be improved.
Light-emitting diodes (LEDs)
As in automobiles, light-emitting diodes will continue to replace traditional light sources. New wavelengths will slowly drop further into the UV, which will lead to improved imaging with fluorescent stains emitting in the visible. The diode’s flicker may be harnessed or possibly used with fast electronics to extend the dynamic range.
Wi-Fi communication in the lab
Wi-Fi communication has not been used in the laboratory. Interconnections between nodes of most instrument systems are hardwired, which is messy at best. Intralaboratory Wi-Fi communication should reduce frustration in reconfiguring instruments for new assays or experiments.
Assays are too often dependent on the instrumentation. Method transfer is an issue. Details are important and seldom discussed. These issues are being addressed. Quality-by-design approaches are defining the important operational limits and design space.
Bio-banking is another automation opportunity. Today, most samples are stored and accessed with 1-D or 2-D barcodes that are applied to the exterior of the container or plate. However, radiofrequency identification (RFI) chips encased in each sample vial appear to offer much greater assurance and convenience.3
In conclusion, a host of products have been introduced that are transforming work flows while bringing much-needed automation to the laboratory. These include systems for confocal scanning, semantic technology, digital dispensing, materials science, microfluidics, and bio-banking.
- Kallenbron, J.; Hantash, J. et al. A Comparison Between a Developed Method for Determination of Insulin in Human Serum Using the Gyros Platform and Traditional ELISA and RIA Assays; www.aapsj.org/abstracts/NBC_2012/T2045.pdf.
- Chen, C.-C.; Letian, D. et al. Visibly Transparent Polymer Solar Cells Produced by Solution Processing; www.pubs.acs.org/doi/ abs/10.1021/nn3029327.
- Davidowitz, H. Use of radio frequency identification (RFID) for sample tracking. Am. Lab.2012, 44(7), 20–3; http://www.americanlaboratory.com/913-Technical-Articles/118171-Use-of-Radio-Frequency-Identification-RFID-for-Sample-Tracking/
Robert L. Stevenson, Ph.D., is a Consultant and Editor of Separation Science for American Laboratory/Labcompare; e-mail: firstname.lastname@example.org.