See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/322186681

Lab-on-a-Chip Technology and Its Applications

Chapter · January 2018

DOI: 10.1016/B978-0-12-804659-3.00008-7

CITATIONS READS

11 859

2 authors, including:

Burak Yilmaz
Kernal Biologics, Cambridge, MA
17 PUBLICATIONS 11 CITATIONS

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

mRNA immunogenicity View project

All content following this page was uploaded by Burak Yilmaz on 04 November 2019.

The user has requested enhancement of the downloaded file.

 C H A P T E R

8
Lab-on-a-Chip Technology and
Its Applications
Burak Yılmaz and Fazilet Yılmaz
Turgut Ozal University, Ankara, Turkey

8.1 INTRODUCTION

A microfluidic platform contains set of well-defined fabricated fluidic operational units.

Lab-on-a-chip (LOC) platforms provide a solid way for miniaturization, integration, auto-
mation, and parallelization of chemical processes (Mark et al., 2010).
The main advantage of LOC devices is cost efficiency because of using fewer reagents
in modular fabricated miniaturized devices. Another important advantage is speed; paral-
lelization of reaction chambers in LOC devices speed up the operation in the small area.
Moreover, LOC technologies increase throughput and automation of analytical systems.
LOC devices can be designed to control fluidics in micro- and nanoscales, and according
to this scale difference sometimes they are called as microfluidics and nanofluidics. In
LOC technology, micro- or nanochannels allow control of fluids in low quantities to enable
biochemical reactions at very small volumes. Integrated circuits and microchannels are
designed by using photolithography method, and it allows creating paths and control ele-
ments for fluids. LOC also requires integrated pumps, electrodes, valves, electrical fields,
and electronics to become complete systems (Casquillas and Timothée, 2015).
Development of integrated circuit technology and wafer fabrication facilities leads the
rise of microfluidics and lead to smaller and faster electronic devices. Microfluidic fabrica-
tion is based on silicon or glass and polymers. Silicon and glass have well-controlled
mechanical and chemical properties, but they need more manufacturing costs and high
processing complexity.
However, polymers can easily be fabricated via soft lithography or hot embossing,
where a single mold can be used as a template for many devices. This enables production
of high-volume disposable devices. By contrast, the mechanical and chemical properties of

Omics Technologies and Bio-engineering: Towards Improving Quality of Life

DOI: https://doi.org/10.1016/B978-0-12-804659-3.00008-7 145 Copyright © 2018 Elsevier Inc. All rights reserved.
146 8. LAB-ON-A-CHIP TECHNOLOGY AND ITS APPLICATIONS

polymers have reliability problem, and it makes surface modification, which is important
for robust device functionality (Neuži et al., 2012).
LOC has been used in amount applications, and molecular biology is the core technol-
ogy for LOC; applications mainly focus on diagnostic and genomic analysis, and other
applications are biochemical analysis, proteomic and cell research, biosensor, and drug
development (Table 8.1).
LOC are being applied in lots of molecular biology experiments such as DNA isolation,
PCR, qPCR, electrophoresis, and sequencing. LOC has huge potential to build high-speed
and more sensitive DNA/RNA amplification on PCR microfluidics systems. During any
pandemic, LOC-PCR devices offer rapid detection of viruses and bacteria to overcome
infection. Scaling down PCR-based diagnostic devices by LOC technologies enables
laboratory-independent genetic diagnostic services such as HIV and HBV infection. These
developing LOC devices can also be used for biomarker identification of genetically based
disease. Moreover, the food industry can use LOC devices for rapid pathogen detection
because it speeds up customs clearance during food transporting (Timothée, 2015a).
DNA sequencing has been used in molecular biology for several decades. The first
human genome project was completed in 15 years. Today, using LOC devices to sequence
a human genome can be completed within hours, which is thousands times faster. Next-
generation sequencing platforms are using LOC technologies to read millions of DNA

TABLE 8.1 LOC Technologies and Applications

Area of LOC Application of LOC Products in Market References

1. Diagnostics DNA isolation, PCR, qPCR, Fluidigm BiomarkHD, Agilent Timothée,

electrophoresis, sequencing LabChip, Elvesys system 2015b
Kaigala
et al., 2010

Beer et al.,
2007

2. Genomics Next-generation sequencing Illumina Hiseq, Ion torrent, Metzker,

microarray Nanopore, PasificBio 2005
Wang, 2000

3. Biochemical Immunological assays, glucose Dexcom G5 Wang et al.,

assays monitoring 2003

4. Proteomics SDS-PAGE, MS 908 Devices Zip Chip Schasfoort,

2004
Mouradian,
2002
5. Biosensors Bioaffinity and biocatalytic devices IST AG IV4 biosensor Wang, 2000

6. Cell research Cell culturing and monitoring, flow Fluidigm Helios Huh et al.,
cytometers 2005

I. EMERGING FIELDS
 8.1 INTRODUCTION 147
fragments in a single chip, most of them are used in laboratories; Nanopore technologies
is one among them that developed the smallest and fastest platform waiting to be
commercialized.
Protein analysis includes cell extraction, electrophoresis, blotting, digestion, and mass
spectrometry (MS) analysis. LOC integrates all these protein analysis steps in one chip.
Integrating steps reduce analysis time into few minutes. Crystallization process can be par-
allelized and speeded up in LOC systems to study their structure. Additionally, immu-
noassays are the other LOC applications to shorten the reaction compared to the
conventional approach. LOC microchannels are the most suitable platform for cell
research; microfluidics enables controlling cell flow, labeled antibody staining and
imaging, cell differentiation, cell sorting, and cytometry in cell biology experiments.

DNA = Adenine + Cytosine + Thymine + Guanine

8.1.1 Diagnostics
LOCs are being applied in lots of molecular diagnostic steps: DNA extraction and puri-
fication, PCR, qPCR, molecular detection, electrophoresis, etc.

8.1.1.1 DNA Extraction and Purification on LOC Devices

Nucleic acid needs to be purified from biological cells for diagnostic assays. DNA
extraction basically requires two steps: cell lysis by disrupting cells membrane and nucleus
membrane, and DNA purification from unwanted samples such as membrane lipids, pro-
teins, and RNA. LOC cell lysis devices can be based on several types of lysis methods:
chemical lysis, thermal lysis, ultrasonic lysis, electrical lysis, mechanical lysis, etc.
Most of the macroscopic DNA purification methods made in extraction columns, using
the DNA adsorption on silica beads under certain buffer conditions, are adaptable to
microfluidic techniques. In LOC devices, silica beads can thus either be blocked by
mechanical obstacles or silica-coated magnetic beads to easily block these particles with a
magnet for the washing and elution phases (Timothée, 2015b).
DNA sequencing means to know the order and arrangements of ACTG
8.1.1.2 PCR, qPCR, and Molecular Detection on LOC Devices
After DNA extraction, the most common analysis is the PCR (Polymerase Chain
Reaction). PCR has lots of applications that are directly and indirectly like sequencing
techniques. One of the direct PCR applications is obviously the amplification of DNA
sequences that helps to make detectable low amounts of DNA (e.g., for pathogen detec-
tion, like bacteria or virus).
The importance of PCR in genomic analysis affects the development of numerous LOC
devices for PCR. Miniaturization of volume and the high surface to volume ratio leads
rapid thermal transfer for rapid and integrated PCR. PCR also requires a post-analysis so
that amplicons’ size detection carried out by electrophoresis have been made to integrate
PCR and electrophoresis on-chip (Timothée, 2015a).
Originally electrophoresis is done by using gels, mainly made using agarose (for longer
DNA) and polyacrylamide (for shorter DNA). With the advent of LOCs, DNA electropho-
resis was one the first molecular processes that could be integrated on a chip (Curtis
Saunders et al., 2013). This miniaturization enabled to increase even more the process time

I. EMERGING FIELDS
148 8. LAB-ON-A-CHIP TECHNOLOGY AND ITS APPLICATIONS

FIGURE 8.1 PCR microfluidic system.

to realize electrophoresis, reduce reagent consumption, and assemble on a chip other

DNA process steps as mentioned before (Fig. 8.1).
qPCR is another technique that was adapted to LOC devices that present the advantage
to be faster (automated detection during PCR), more sensitive, and sustainable.
As an example, using ultra-fast pressure controller and fluorescence reader and based
on the ultra-fast temperature control, ultra-fast qPCR microfluidic system had been devel-
oped by Elvesys system for the molecular detection of diseases like Anthrax and Ebola in
less than 8 minutes with a detection efficiency identical to commercial systems that are
7 15 times slower (Ramalingam et al., 2010).
Another emerged field is digital microfluidics that deals with emulsion and droplets
within LOC devices.
Ultra-low amount of DNA can be captured within droplets, and limits can be increased
with one copy number detection within LOC droplet qPCR (Beer et al., 2007) (Fig. 8.2).

8.1.2 Genomic Application

The field of genomics has had a tremendous growth rate in recent years because of the
effects of sequencing projects such as the Human Genome Project. The projects related to
sequencing accelerate the sequencing technologies. Sequencing is the analysis determining
the nucleotide order of DNA sequences. The first sequencing was developed based on the
Sanger method; in this method, DNA fragments with different lengths and different fluor-
ophores corresponding to different nucleotide ends will be obtained. Miniaturization of
the standard DNA sequencing, using a slab of polyacrylamide gel, was achieved by repla-
cing the gel slab with a gel-filled small capillary. The small dimensions of the capillaries
decreased the required DNA sample volume (Figeys and Pinto, 2000).

I. EMERGING FIELDS
 8.1 INTRODUCTION 149

FIGURE 8.2 LOC droplet qPCR.

TABLE 8.2 Comparison of Time and Cost for Next-Generation Sequencing Systems That Use
LOC Technologies

Runtime Read Length Output Nucleotide Cost/MB

Illumina Miseq 27 h 2 3 300 bp 15 GB $0.15

Illumina Hiseq 2500 1 10 days 2 3 250 bp 3000 GB $0.05

Ion Torrent 2h 400 bp 2 GB $1

Pacific Biosciences 30 min 4 h 10 40 kb 500 1000 MB SMRT cell $0.13 $0.80
Nanopore Technologies ,48 h 5 200 kb 50 100 GB $0.003 $0.01

Next-generation sequencing for ultra-fast DNA sequencing currently under development

include sequencing-by-hybridization (SBH), nanopore sequencing, and sequencing-by-
synthesis (SBS), the latter of which encompasses many different DNA polymerase-
dependent strategies. Using LOC approach for these technologies enables high-throughput
sequencing to reduce drastically its cost and duration (Table 8.2). Most famous next-
generation sequencing systems are Illumina and its sequencing by synthesis technology,
Life technologies and its Ion Torrent using semiconductor technologies, Pacific Bio with its
single molecule real-time sequencing using zero-mode waveguides, and Roche with its
454 sequencing system using pyrosequencing technology (Metzker, 2005).

8.1.3 Microarray
The analysis of complex DNA samples expression needs integration of multiple biosen-
sors in connection with DNA microarrays. The most important features of these LOCs are
miniaturization, speed, and accuracy. This technology offers huge potential for rapid mul-
tiplex analysis of nucleic acid samples, including the diagnosis of genetic diseases, detec-
tion of infectious agents, measurements of differential gene expression, drug screening, or
forensic analysis. Such use of DNA microarrays is thus revolutionizing many aspects of
genetic analysis (Wang, 2000).

I. EMERGING FIELDS
150 8. LAB-ON-A-CHIP TECHNOLOGY AND ITS APPLICATIONS

8.1.4 Biochemical Applications

LOC strategy for coupling enzymatic and immunological assays on a single-channel
microfluidic device is also applicable, for example simultaneous glucose and insulin mea-
surements. The availability of an LOC capable of simultaneously monitoring both insulin
and glucose holds great promise for improved management of diabetes.
Crucial for the successful realization of such glucose/insulin monitoring is the integra-
tion of relevant sample pretreatment/cleanup procedures essential for whole-blood
analysis. The new LOC approach can be applicable to the integration of other assays
and additional sample handling steps, as is desired for creating miniaturized clinical
instruments (Wang et al., 2003).

8.1.5 Proteomics
Proteomics is one of the great scientific challenges in the post-genome era. The most
basic form of proteomics is proteome profiling, identifying all the proteins expressed in
each sample, which is a demanding task. The proteome has unique analytical challenges,
including molecular diversity, wide concentration range, and a tendency to adsorb to solid
surfaces.
LOC devices are useful for developing new methods to solve complex analytical pro-
blems, such as proteome profiling. LOC devices seem to be progressive in four key areas
related to this application: chemical processing, sample preconcentration and cleanup,
chemical separations, and interfaces with mass MS (Freire and Wheeler, 2006).
LOC is a miniaturized device suited for separation and detection of proteins which
enables less reagent consumption, easy operation, and very fast analysis. Because aim is to
obtain the maximum amount of data from each sample, LOC protein devices can generate
a huge amount of data points. Another advantage is that the data from the LOC devices
can easily be compared to those obtained using 2D-PAGE (Schasfoort, 2004).
LOC protein sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)
separation with fluorescence detection is the most commonly used method to separate and
size protein mixtures. Normally, SDS-PAGE is time consuming and labor intensive. LOC
devices speed up separation and lay the building blocks toward automation of protein
sizing (Mouradian, 2002).
LOC MS protein profiling is one of the applications to quantitate proteins.
Mouradian (2002) describes LOC devices that are used for direct infusion into the mass
spectrometer, including Capillary Electrophoresis (CE) separation, on-chip sample digestion,
and infusion or CE before MS analysis. Researchers use electrospray ionization MS and
focus on the interface design between LOC and the mass spectrometer (Mouradian, 2002).

8.1.6 Biosensors
LOC biosensors are small devices in which biological reactions occur for detecting
target analytes. Such devices closely link together a biological recognition element (inter-
acting with the target analyte) with a physical transducer converting the biorecognition
event into an electrical signal. There are two types of LOC biosensors, which are

I. EMERGING FIELDS
 8.1 INTRODUCTION 151
bioaffinity and biocatalytic devices. Bioaffinity devices are based on selective binding of
the target analyte to a surface-enclosed ligand partner (e.g., antibody, oligonucleotide).
For example, in hybridization biosensors, there are immobilized single-stranded (ss)
DNA probes onto the transducer surface. The duplex formation can be detected follow-
ing the association of an appropriate hybridization indicator after binding. On the other
way, in biocatalytic devices, an immobilized enzyme is used for recognizing the target
substrate. For example, sensor strips with immobilized glucose oxidase for monitoring
of diabetes (Wang, 2000).

8.1.7 Cell Research

LOC devices enable the researcher to handle small-scale experiments with cells. In
many cases, the single cell, when positioned within the microchannel, will also need to be
positioned or trapped in a predetermined location, making single-cell handling and
manipulation necessary. The microsystem should be able to handle small volumes of fluid
containing small quantities of analyte. LOC systems also provide an ability to control the
local environment around the cell with precision, using methods that include having con-
trol over the physical or chemical nature of the surface to which the cell adheres, the local
pH, and temperature. When the cell is to be exposed to a different kind of stimuli, multi-
ple and often complex fluid-handling components can be used. Together, these ideas
imply the need for the integration of cell positioning and chemical stimulation. If the cell
is then to be analyzed, lysis and analysis should be introduced onto the same platform to
avoid sample loss or dilution, which would inevitably occur if multiple devices were used
(Sedgwick et al., 2008).
Flow cytometers are one of the successful instruments in the earlier stage and the most
useful among LOC devices developed to date. In the past decade, LOC systems of flow
cytometers have been developed rapidly by two main factors: (1) the increased need for
higher quality and larger quantity of cellular analysis data, and (2) the enhanced sophisti-
cation and accessibility of microfabrication technologies. The emerging needs coupled
with the new technical capabilities have led to a variety of exciting developments in micro-
fluidics for flow cytometric analysis of cells and particles (Huh et al., 2005).

8.1.8 Drug Development

The industry needs new tools to guide the development of new drugs — especially to
help predict the behavior of potential new drugs in humans from performance in animals
and cells. Some analytical applications of LOC systems in the production and use of bio-
pharmaceuticals seem straightforward (e.g., analytical systems to monitor and optimize
the production of protein drugs such as therapeutic antibodies); others (such as assays
based on primary human cells that could predict performance in human clinical trials) are
technically more complicated, but also feasible, at least in some instances. In either case,
LOC systems in a highly reproducible and easily manipulated format could be used
routinely (Whitesides, 2006).

I. EMERGING FIELDS
152 8. LAB-ON-A-CHIP TECHNOLOGY AND ITS APPLICATIONS

8.2. CONCLUSION AND FUTURE DIRECTIONS

Miniaturization of laboratory devices into chip-sized devices is a new area called LOC.
Cost, time, and parallelization are main advantage of this field, and it integrates several
operations with better quality of sensitivity.
By looking into commercialized LOC devices such as PCR, DNA sequencing, and glu-
cose monitoring, we can say that LOC will change diagnostic concepts in the near future.
And we will see more products in the market developed using LOC technologies. The
industry will reach more economical production of LOC devices day by day. LOC devel-
opment directly affects molecular-based innovations to be used in daily life.

References
Beer, N.R., Hindson, B.J., Wheeler, E.K., Hall, S.B., Rose, K.A., Kennedy, I.M., et al., 2007. On-chip, real-time,
single-copy polymerase chain reaction in picoliter droplets. Anal Chem 79 (22), 8471 8475. Available from:
https://doi.org/10.1021/ac701809w.
Casquillas, G., Timothée, H., 2015. Introduction to lab-on-a-chip 2015: review, history and future. Retrieved from
http://www.elveflow.com/microfluidic-tutorials/microfluidic-reviews-and-tutorials/introduction-to-lab-on-a-
chip-2015-review-history-and-future/.
Curtis Saunders, D., Holst, G.L., Phaneuf, C.R., Pak, N., Marchese, M., Sondej, N., et al., 2013. Rapid, quantitative,
reverse transcription PCR in a polymer microfluidicchip. Biosens Bioelectron 44, 222 228. Available from:
https://doi.org/10.1016/j.bios.2013.01.019.
Figeys, D., Pinto, D., 2000. Lab-on-a-chip: a revolution in biological and medical sciences. Anal Chem 72 (9),
330A 335A. Available from: https://doi.org/10.1021/ac002800y.
Freire, S.L.S., Wheeler, A.R., 2006. Proteome-on-a-chip: mirage, or on the horizon? Lab Chip 6 (11), 1415.
Available from: https://doi.org/10.1039/b609871a.
Huh, D., Gu, W., Kamotani, Y., Grotberg, J.B., Takayama, S., 2005. Microfluidics for flow cytometric analysis of
cells and particles. Physiol Meas 26 (3), R73 R98. Available from: https://doi.org/10.1088/0967-3334/26/3/
R02.
Kaigala, G.V., Behnam, M., Bidulock, A.C.E., Bargen, C., Johnstone, R.W., Elliott, D.G., et al., 2010. A scalable and
modular lab-on-a-chip genetic analysis instrument. Analyst 135 (7), 1606 1617. Available from: https://doi.
org/10.1039/b925111a.
Mark, D., Haeberle, S., Roth, G., von Stetten, F., Zengerle, R., 2010. Microfluidic lab-on-a-chip platforms: require-
ments, characteristics and applications. Chem Soc Rev 39 (3), 1153 1182. Available from: https://doi.org/
10.1039/b820557b.
Metzker, M.L.M.L.L., 2005. Emerging technologies in DNA sequencing. Genome Res 15 (12), 1767 1776.
Available from: https://doi.org/10.1101/gr.3770505.with.
Mouradian, S., 2002. Lab-on-a-chip: applications in proteomics. Curr Opin Chem Biol 6 (1), 51 56. Available
from: https://doi.org/10.1016/S1367-5931(01)00280-0.
Neuži, P., Giselbrecht, S., Länge, K., Huang, T.J., Manz, A., 2012. Revisiting lab-on-a-chip technology for drug dis-
covery. Nat Rev Drug Discov 11 (8), 620 632. Available from: https://doi.org/10.1038/nrd3799.
Ramalingam, N., Rui, Z., Liu, H.B., Dai, C.C., Kaushik, R., Ratnaharika, B., et al., 2010. Real-time PCR-based
microfluidic array chip for simultaneous detection of multiple waterborne pathogens. Sensors Actuat B Chem
145 (1), 543 552. Available from: https://doi.org/10.1016/j.snb.2009.11.025.
Schasfoort, R.B.M., 2004. Proteomics-on-a-chip: the challenge to couple lab-on-a-chip unit operations. Expert Rev
Proteomics 1 (1), 123 132. Available from: https://doi.org/10.1586/14789450.1.1.123.
Sedgwick, H., Caron, F., Monaghan, P.B., Kolch, W., Cooper, J.M., 2008. Lab-on-a-chip technologies for proteomic
analysis from isolated cells. J R Soc Interface 5 (Suppl 2), S123 S130. Available from: https://doi.org/10.1098/
rsif.2008.0169.focus.
Timothée, H., 2015a. Microfluidic PCR, qPCR, RT-PCR & qRT-PCR. Retrieved from http://www.elveflow.com/
microfluidic-tutorials/microfluidic-reviews-and-tutorials/microfluidic-pcr-qpcr-rtpcr/.

I. EMERGING FIELDS
 FURTHER READING 153
Timothée, H., 2015b. Microfluidics for DNA analysis. Retrieved from http://www.elveflow.com/microfluidic-
tutorials/microfluidic-reviews-and-tutorials/microfluidics-for-dna-analysis-pcr/.
Wang, J., 2000. From DNA biosensors to gene chips. Nucleic Acid Res 28 (16), 3011 3016. Available from:
https://doi.org/10.1093/nar/28.16.3011.
Wang, J., Ibáñez, A., Chatrathi, M.P., 2003. On-chip integration of enzyme and immunoassays: simultaneous mea-
surements of insulin and glucose. J Am Chem Soc 125 (28), 8444 8445. Available from: https://doi.org/
10.1021/ja036067e.
Whitesides, G.M., 2006. The origins and the future of microfluidics. Nature 442 (7101), 368 373. Available from:
https://doi.org/10.1038/nature05058.

Further Reading
Blazej, R.G., Kumaresan, P., Mathies, R.A., 2006. Microfabricated bioprocessor for integrated nanoliter-scale
Sanger DNA sequencing. Proc Natl Acad Sci U S A 103 (19), 7240 7245. Available from: https://doi.org/
10.1073/pnas.0602476103.

I. EMERGING FIELDS

View publication stats