
Lab-on-a-Chip
Other Unique Engineering Ideas
An active
field is the integration of the sample preparation and DNA array
detection in the so-called ‘Lab-on-a-Chip’ configuration. The goal of
this technology is to fully integrate multiple processes, including
sample collection and pretreatment with the DNA extraction,
amplification, hybridization and detection, on a microfluidic platform.
1. Description
2. Why
3. How
4. Future Trends
5. Related Links
Description
The ability to perform all the steps of the biological assay on a single self-contained microchip promises significant advantages in terms of speed, cost, sample/reagent consumption, contamination, efficiency and automation (including parallel sample processing).Such miniaturization of the analytical instrumentation will enable transportation of the laboratory to the sample source (as desired for point-of-care testing). Sophisticated devices have thus been fabricated with pumps, valves, heaters, filters, along with the corresponding fluidic network.For example, PCR amplification has been performed by continuously flowing the sample through three well-defined temperature controlled zones on a glass microchip. The pattern of the chip layout determined the relative time a fluid element is exposed to each temperature zone. An array of PCR microchips, based on multiple reaction chambers with resistive heaters was developed at the Lawrence Livermore National Laboratory. A microchip device for cell lysis, multiplex PCR and electrophoretic sizing has also been described. Voltages, applied through electrodes placed within the individual reservoirs, are used to drive the electroosmotic flow. Such electroosmotic ‘pumping’ obviates the need for mechanical pumps or valves, with the channel intersections serving as ‘virtual injection valves’.
Why
Lab-on-a-Chip enables sample handling, mixing, dilution,
electrophoresis and chromatographic separation, staining and detection
on single integrated systems. The purpose of these devices is to
manipulate and process solution based samples and systems by carrying
out typical procedures such as mixing, heating and separation.
Processed samples may be delivered to some form of detector that
subsequently transmits data. It is far less expensive than conventional
methods of analyzing DNA (which require specialized laboratories,
equipment, and personnel) yet just as quick and sensitive.The main
advantages of Lab-on-a-Chip are
- ease-of-use,
- speed of analysis,
- low sample and reagent consumption and
-
high reproducibility due to standardization and automation
Microfabricated devices are also attractive for high-throughput electrically-driven DNA separations. For example, Woolley et al.
described capillary electrophoresis microchips that allow DNA sizing of
12 samples in parallel with a resolution of 10 bp and higher
throughputs than conventional techniques.
How
These
devices typically consist of a monolithic material that is patterned
with microscale channels and features such as mixers, valves, injectors
and separators that can assume the role of equivalent macroscopic
laboratory equipment. The preparation of these credit-card sized
microlaboratories commonly relies on advanced microfabrication and
micromachining technologies, using processes common in the manufacture
of electronic circuitry.Appropriate connection to the macroscale world
for inputs and outputs (not trivial) is required and so the
microfluidic device is likely to be part of a system. Controlled
electric fields have been used for discriminating among oligonucleotide
hybrids with varying binding strengths (including between complete
match and single-base mismatch) and to expel the unhybridized DNA. Such
fine-tuned electronic stringency selection obviates the need for
extensive washing. The electronically-regulated sample preparation
process was demonstrated for the dielectrophoretic separation of
Eschericia coli from blood cells. After the isolation, the bacteria
were lysed by a series of high-voltage pulses.Routine
processing of samples and microreactors for selected chemical reactions
are likely to represent some of the earlier applications of
microfluidic devices to allow speed, control and reduced cost compared
to incumbent solutions. Although the advances in genomics (in which DNA
microarrays have played a key role) have been crucial and will continue
to play a vital role they only represent part of the life science
puzzle.
Future Trends
On-chip fluid manipulations have been demonstrated for sequentially transporting nanoliter samples through a network of microchannels, mixing of sample and reagents, dispensing of samples, DNA restriction digestion and electrically-driven separations. A variety of microstructures have been proposed for on-chip PCR amplification.The first commercially available product based on microfluidic technology is the HP 2100 Bioanalyzer (Hewlett Packard). This instrument integrates the sample handling and electrophoretic separation.Nanogen Inc. (San Diego, CA) has developed an electronic sample preparation process. The company is addressing each step in the sample-to-result process on microfabricated chip-based devices. This includes the integration of electronic cell separation, electronic sample transport, electronically-accelerated hybridization and electronic denaturation.
Keywords
DNA Analysis, Microarrays, Lab-on-a-Chip, Microfluidic Lab-on-a-Chip.
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