Anal. Chem.
2008, 80, 9131–9134
Paper-Based Microfluidic Devices by Plasma
Treatment
Xu Li, Junfei Tian, Thanh Nguyen, and Wei Shen*
Australian Pulp and Paper Institute, Department of Chemical Engineering, Monash University, Wellington Road,
Clayton, Victoria 3800, Australia
Paper-based microfluidic patterns have been demon-
strated in recent literature to have a significant potential
in developing low-cost analytical devices for telemedicine
and general health monitoring. This study reports a new
method for making microfluidic patterns on a paper
surface using plasma treatment. Paper was first hydro-
phobized and then treated using plasma in conjunction
with a mask. This formed well defined hydrophilic chan-
nels on the paper. Paper-based microfluidic systems
produced in this way retained the flexibility of paper and
a variety of patterns could be formed. A major advantage
of this system is that simple functional elements such as
switches and filters can be built into the patterns. Ex-
amples of these elements are given in this study.
This article reports on a novel method of creating patterns in
paper to control the transport of liquid for pathological testing
and environmental monitoring. Nonwoven fibrous materials such
as paper have very good potential as low-cost base materials for
diagnostic devices for such applications. Cellulose-based papers
have been used as chromatographic substrates since 1850.1,2
A number of testing devices, such as paper-based indicators3 and
“dip stick” test assays are made of paper. Recently, Martinez et
al.4-6 used paper as a base material to create barrier patterns with
photolithographic techniques. The photoresist patterns they
produced guide the capillary penetration of liquids and, with the
appropriate indicators, can detect chemical components in a liquid
sample. This type of paper-based detector, when used with mod-
ern communication tools such as camera phones, can provide
powerful real-time and off-site diagnostic devices for telemedicine
with great potential to be developed into a low-cost health
monitoring system.5 The drawback of conventional photoresist
* Corresponding author. E-mail: wei.shen@eng.monash.edu.au. Phone: +61
3 99053447. Fax: +61 3 99053413.
(1) Stock, R.; Rice, C. B. F. Chromatographic Methods, 3rd ed.; John Wiley &
Sons: New York, 1974; p 106.
(2) Atkins, P.; Jones, L. Chemistry-Molecules, Matter, and Change, 3rd ed.; W.H.
Freeman & Company: New York, 1997; p 26.
(3) Umland, J. B. General Chemistry; West Publishing Company: St. Paul, MN,
1993; p 592.
(4) Martinez, A. W.; Phillips, S. T.; Buttle, M. J.; Whitesides, G. M. Angew. Chem.,
Int. Ed. 2007, 46, 1318–1320.
(5) Martinez, A. W.; Phillips, S. T.; Carrilho, E.; Thomas, S. W., III; Sindi, H.;
Whitesides, G. M. Anal. Chem. 2008, 80, 3699–3707.
(6) Martinez, A. W.; Phillips, S. T.; Wiley, B. J.; Gupta, M. J.; Whitesides, G. M.
Lab Chip 2008, DOI: 10.1039/b811135a.
10.1021/ac801729t CCC: $40.75  2008 American Chemical Society
Published on Web 11/01/2008
is that it is much less flexible than paper, and the device can be
easily damaged by bending and folding.7
More recently, Bruzewicz et al.7 used a modified plotter to
create a barrier pattern by printing polydimethylsiloxane (PDMS)
onto paper. When PDMS penetrated into paper, it formed a barrier
pattern and could be used to control the penetration of liquid in
channels defined by the barrier pattern. Since PDMS is an
elastomer, it is more flexible than conventional photoresist and
is less susceptible to mechanical damage. However, because of
the low surface tension of the solvent used, the PDMS solution
penetrates into paper rather easily. Since paper is not a uniform
porous material, the penetration of PDMS cannot be well con-
trolled, resulting in the wall of the barrier pattern not being
straight.
The novel method of the present study enables patterns to be
created on paper surfaces without affecting their flexibility or
surface topography. The major advantage of this novel method
over previously reported paper-based microfluidic devices is that
it allows not only sample testing but also building of simple
functional components such as control switches, microfilters, and
microreactors, which are important elements for multiple-step
chemical tests. Several examples of these functional elements are
presented in this work.
EXPERIMENTAL SECTION
Alkyl ketene dimer (AKD) (Wax 88 konz, BASF) was used as
the cellulose hydrophobization agent. Analytical grade n-heptane
(Aldrich) was used as the solvent for AKD. Whatman filter paper
was selected as the paper substrate. MilliQ water was used to
prepare all liquid samples required for testing the performance
of the device.
The filter paper was hydrophobized using an AKD-heptane
solution (0.6 g/L). The filter paper samples were dipped in this
solution and immediately removed and placed in a fume cupboard
to allow evaporation of the heptane. The filter paper samples
were then heated in an oven at 100 °C for 45 min to cure the
AKD. The treated filter paper samples are strongly hydrophobic
and have contact angles of typically 110-125° with water.9
Therefore, water shows no penetration into the paper hydropho-
bized with AKD.
(7) Bruzewicz, D. A.; Reches, M.; Whitesides, G. M. Anal. Chem. 2008, 80,
3387–3392.
(8) Shen, W.; Tian, J.; Li, X.; Garnier, G. Australian Provisional Patent No.
2008903553, July 11, 2008.
(9) Shen, W.; Filonanko, Y.; Truong, Y.; Parker, I. H.; Brack, N.; Pigram, P.;
Liesegang, J. Colloids Surf., A 2000, 173, 117–126.
Analytical Chemistry, Vol. 80, No. 23, December 1, 2008 9131
Figure 1. Water penetration control of a plasma treated microfluidic
channel using hydrophobized filter paper. (a) A hydrophobized filter
paper sample immediately after the treatment and (b) the treated
sample was exposed to water through the lower edge.
Hydrophilic patterns were formed on paper samples using
plasma treatment. The paper samples were sandwiched between
metal masks having the desired patterns and then placed into a
vacuum plasma reactor (K1050X plasma asher (Quorum Emitech,
U.K.)) for 15 sec at the intensity of 15 W. Metal masks were made
by mechanically cutting patterns through stainless steel sheets.
Four screws were used to secure the alignments of the masks.
The vacuum level for the treatment was 6 × 10-1 mbar. The plasma
treatment left no visible mark on the samples, which retained its
original flexibility. The plasma treated areas were strongly wettable
by water or aqueous solutions and allowed the transport of
aqueous solutions along and within the plasma treated channels
via capillary penetration.8
A color change reaction for the evaluation of enzyme activity
was used to test the device. Alkaline phosphatase enzyme solution
was prepared using alkaline phosphatase (lyophilized powder,
from bovine intestinal mucosa, 10-30 DEA units/mg of solid)
from Sigma-Aldrich. Phosphatase solution (1 mg/mL) was pre-
pared in the buffer solution (pH ) 9.8) which contains 1.0 M
diethanolamine (g98%, Sigma Aldrich) and 0.5 mM MgCl2 ·
6H2O (g99%, Sigma-Aldrich). The phosphatase enzyme solution
was divided into two parts; one part was heated at >70 °C for 10
min to purposely deactivate the enzyme. A BCIP/NBT substrate
system (Sigma-Aldrich) was used with the phosphatase enzyme
to indicate the activity of the enzyme via color change. A
phenolphthalein-ethanol solution was used as the color indicator
for testing the performance of a paper-based microfluidic reactor.
It was prepared using phenolphthalein (BDH) and ethanol
(AR, BDH).
RESULTS AND DISCUSSION
Plasma Formed Patterns. Parts a and b of Figure 1 show a
filter paper sample immediately after treatment and after being
exposed to water. Water penetration demonstrated that the plasma
treated channel has well-defined borders; the width of the channel
can be reasonably well controlled. The channel in Figure 1b was
made using a 1 mm wide mask, and a channel <1.5 mm in width
was uniformly penetrated by water.
Hot electrons and other energetic particles generated in
vacuum plasma have long mean free paths. Overetching of the
substrate under a mask is a known problem for plasma treatment,
9132 Analytical Chemistry, Vol. 80, No. 23, December 1, 2008
Figure 2. The performance of a paper-based microfluidic pattern
fabricated by plasma treatment. Three active and three deactivated
enzyme samples were introduced into detection ports of a six-channel
pattern in alternation. A few drops of liquid substrate (BCIP/NBT) were
introduced from the center port. Liquid substrate penetrated into the
detection ports, revealing the activity of the enzyme samples.
causing the treated pattern to be slightly bigger than the mask.
However, overetching could be controlled by optimizing the
treatment intensity and time. When the treatment intensity and
time were fixed, the channel width was found very reproducible.
Design of Sample Dosing and Detection Sites on Paper-
Based Microfluidic Devices. An important application of paper-
based microfluidic devices is to analyze chemical components in
a liquid sample by means of color indicators.4-7 The research
group in Harvard University was the first to explore this
application.4-7 In their approaches, the concept of a capillary
channel barrier was used to confine the liquid penetration path
from sample dosing zone(s) to sample detection zones. In our
approach, plasma generated hydrophilic patterns in a hydropho-
bized paper can control the capillary flow path of liquids and can
be designed to have one or more detection zones. Figure 2 shows
a test of a plasma generated pattern. Small quantities (2 µL) of
three active and three deactivated enzyme samples were intro-
duced in alternation into detection ports of a six-channel pattern.
A few drops of liquid substrate (BCIP/NBT) were introduced from
the center port of the pattern. Three samples with active enzyme
show a color change, and three deactived enzyme samples show
no color change. This microfluidic system created by plasma
treated paper is capable of performing analytical tests proposed
by other researchers.4-7
Building Functional Elements on Paper-Based Microflu-
idic Systems. Functional elements such as switches, filters, and
separators can be easily built into microfluidic devices made by
plasma treatment of paper. An advantage of plasma treated paper
is that the untreated areas remain hydrophobic and do not allow
capillary penetration of sample liquids. Such structure allows
simple switches to be built so as to connect and disconnect the
microfluidic channels. Several of many possible mechanical
designs can be found in the Supporting Information. Simple switch
Figure 3. A simple and versatile design of a paper-based microfluidic
separator. It can be activated by pulling the paper strip.
Figure 4. A filter made of silica coated matt ink jet paper was used
to separate the magenta dye from a diluted ink jet ink solution.
Figure 5. A design of a simple paper-based microfluidic reactor
consisting of two sample dosing sites (A1, A2), two switches (S1, S2),
and a reaction site (B).
design can be found below (Figures 5 and 6). It is possible that
such switches can be activated by some other means such as by
electrostatic or electromagnetic forces, etc.
A filter or a separator can also be easily designed for paper-
based microfluidic systems.With dependence on the requirement
of the analytical task, ion-exchange resins, high surface area
functionalized nano- or microparticles or cationic polymer-coated
mineral fillers can be deposited on the surface of the mechanical
Figure 6. A microfluidic reactor in operation: (a) The reactor
fabricated using plasma treatment. Phenolphthalein was deposited
in the reaction site B. (b) Dosing of a NaOH solution via A1 and HCl
solution via A2; (c) NaOH solution was allowed into the reaction site,
triggering indicator color change; (d) HCl solution was allowed into
the reaction site, causing indicator color fading; (e) complete reversion
of indicator color by HCl solution.
switches. When the switch is in the connected state, the sample
liquid can be filtered to remove solid particles when penetrating
through the filter. Also, certain molecular species can be separated
from the sample liquid chromatographically as the sample
penetrates through a separator.
Figure 3 shows an even simpler and versatile design of a paper-
based chromatographic separator. A strip of paper or other flexible
material with a patch of coating, as shown in Figure 3, can be
used to form the separation element. The separator is activated
by pulling the strip as illustrated in Figure 3. To demonstrate the
operation and effectiveness of this type of separator, a strip of
matt ink-jet printing paper with a silica coating layer was used
to separate a diluted ink-jet ink solution (Figure 4). The diluted
ink was deposited on the right side of the microfluidic channel;
as the solution passed through the separator and reached the left
side channel, dye in the solution was retained by the ink jet paper
separator. Figure 4 indicates that this separator performed
satisfactorily.
With the functional components shown above, a controllable
microfluidic reactor can be built on paper or other suitable
nonwoven materials. Figure 5 shows the concept of a simple
reactor consisting of two sample dosing sites (A1 and A2), which
are connected to a reaction site (B) via two switches (S1 and S2).
Two liquid reactants can be placed in each of the dosing sites;
they can be controlled to enter the reactor by the switches.
Figure 6 shows a microfluidic reactor (Figure 6a) in operation.
A small quantity of phenolphthalein indicator was placed into the
reaction zone (B) to demonstrate an acid-base neutralization
reaction. A NaOH and a HCl solution were introduced into sample
dosing zones A1 and A2, respectively (Figure 6b). Both switches
were in the disconnected position. Switch S1 was then switched
on to allow the NaOH solution to enter the reaction zone. As the
NaOH solution entered the reaction zone, the change of indicator
color was observed (Figure 6c). Then S2 was switched on to allow
the HCl solution to enter the reaction zone (Figure 6d). As the
HCl solution entered the reaction zone, the neutralization reaction
occurred. Figure 6e shows the expected fading of indicator color
as the neutralization reaction completed.
Paper-based microfluidic systems fabricated using plasma
treatment are capable of allowing single- and multi-step tests, as
Analytical Chemistry, Vol. 80, No. 23, December 1, 2008 9133
well as reactions to be performed. Plasma treatment allows the
paper to retain its original flexibility. Since the treatment does
not change the color of the paper, diagnostic devices made using
this method allow easy color identification.
CONCLUSION
A new method of making paper-based microfluidic patterns
by plasma treatment was developed. Microfluidic devices made
using this method have the same capability of transferring and
analyzing liquid samples as similar devices reported previously,
but the paper devices made using plasma treatment have an
advantage over the barrier design in that simple functional
elements such as switches, filters, and separators can be easily
built in the microfluidic system. This advantage will be further
explored to build more advanced paper-based microfluidic devices.
9134 Analytical Chemistry, Vol. 80, No. 23, December 1, 2008
ACKNOWLEDGMENT
The authors would like to gratefully acknowledge Monash
University and Australian Research Council for their scholarships
(X.L. and T.N.). The authors would also like to thank Dr. N.
Cowieson of Monash Centre for Synchrotron Science for valuable
discussions.
SUPPORTING INFORMATION AVAILABLE
Additional information as noted in text. This material is
available free of charge via the Internet at http://pubs.acs.org.
Received for review August 18, 2008. Accepted October 8,
2008.
AC801729T