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DOI 10.1007/s10404-004-0012-9
R EV IE W
Received: 2 August 2004 / Accepted: 18 September 2004 / Published online: 8 April 2005
Springer-Verlag 2005
1 Introduction
Nanouidics is often dened as the study and application of uid ow in and around nanosized objects.
Actually, nanouidics is not new, but has been
aroundwithout a name of its ownfor quite a number
of years, being implicit in many disciplines. The fact that
this previously implicit science of nanoscale uid ow
recently received a name of its own is, however, due to
the rise of microuidics in the 1990s and the recent advent of nanoscience and nanotechnology. The invention
and wide availability of many new technological tools
like atomic force microscope (AFM) and scanning tunnelling microscope (STM) (both for inspection and
creation of nanostructures), the electron, x-beam and
J. C. T. Eijkel (&) A. van den Berg
BIOS/Lab-on-a-Chip Group,
MESA+ Research Institute,
University of Twente,
P.O. Box 217,
7500 AE Enschede,
Netherlands
E-mail: j.c.t.eijkel@utwente.nl
Tel.: +31-534-892839
Fax: +31-534-892287
ion-beam lithographs, and the development of new micromachining techniques like soft lithography and bottom-up assembly methods has made the study and
application of nanouidics much more accessible, and
allows a previously unknown measure of control on the
nanoscale. Expectations of what we can expect from this
new eld are, therefore, high.
While looking forward to future fruits, it is important
not to forget the roots of nanouidics, which it has in
many other disciplines. How this may easily happen is
illustrated by the recent rediscovery of two classical
colloid chemistry phenomena, co-ion (Donnan) exclusion (Pu et al. 2004) and semipermeability (Daiguji et al.
2004), in experiments with and simulations for nanochannels. The authors, clearly, were not aware of the
classical literature on these subjects, which shows how
easily it can be overlooked by nanouidic investigators
exploring their new eld. However, it is of great help to
depart from the knowledge already gathered. Another
reason to start with the roots is to better be able to
identify the places where there is growing potential for
modern nanouidics. This could, for example, be the
case if there is a new and still unexplored area of science
or a new area of applications that can only now be
opened up by using the newly available nanotechnological tools. The impressive array of nanouidic
applications that already exists, and which are described
at the end of this paper, certainly, has resulted from the
application of these new tools, and it is quite reasonable
to expect that there is a lot more to follow.
Figure 1 shows a schematic diagram indicating a
great number of classical disciplines (and one recent one)
where knowledge on nanouidics can be found. For
each discipline, some phenomena studied relevant for
nanouidics are mentioned. The disciplines represent an
enormously diverse array, demonstrating the multidisciplinary approach that is needed for nanouidics research. The diversity of the elds also presented the
authors with a big problem in writing this paper, since
the eld is just too broad to represent in a single papereven a whole book would not be enough. We
250
Fig. 1 Classical disciplines
related to nanouidics and
some of the relevant subjects
studied
venture into a bit of extrapolation and crystal ball gazing to identify possible new applications of nanouidics.
2 Nanoscale forces
Many forces are generally distinguished that play a role
on the nanoscale (Israelachvili 1992a, 1992b). They can
be exerted by walls on particles or solvent molecules or
nearby walls, or by the particles and molecules on each
other. These forces will govern the behaviour of the
molecules or particles in nanostructures, and nanouidic
theory has, therefore, to start with a discussion of these
forces. They can give rise both to equilibrium phenomena, like dierences in ionic distribution, or kinetic
phenomena, like (macroscopic) viscosity. In his excellent
book on intermolecular and surface forces, Israelachvili
remarks that, although our distinction of these forces in
the end is somewhat articial, since all of them are
electrical in nature, such distinction still makes a lot of
practical sense because of the many dierent ways in
which the electrical force presents itself (Israelachvili
1992b). Figure 2 gives a schematic representation of a
number of forces acting on the nanoscale, expressed for
the case of interaction between a spherical particle representing an AFM tip and a at surface, which is taken
from an introductory paper on AFM (Heinz and Hoh
1999). The approximate spatial extension of the forces,
generally, is as indicated in the forcedistance curves, i.e.
reaching furthest for the brush and electrostatic forces,
and the least far for the van der Waals forces.
251
252
b
Fig. 2ad Examples of AFM force curves and the force laws used
to interpret them. Notice that the curves are shown as force versus
sample position curves, which is the most common way in which
they are displayed in literature. However, the force laws are all
functions of the separation distance, D. ac Approach curves. d
Retraction curve. a An ideal, attractive, van der Waals force in the
absence of other forces (Israelachvili 1992b). b Repulsive electrostatic double-layer force in solution (Israelachvili 1992b). c
Polymer-brushing forces that result from the thermally driven
motion of polymers grafted onto a solid surface in solution
(Israelachvili 1992b). Capillary adhesion (very common under
ambient conditions, under which many surfaces have thin layers of
water) results from the formation of a water bridge between the tip
and sample (Israelachvili 1992b). Symbols used: A Hamaker
constant, D probesample separation distance, R probe sphere
radius, L brush thickness in a good solvent, k Boltzmanns
constant, T absolute temperature, s mean distance between
polymers, e dielectric of the medium, k Debye length of the
medium, rR surface charge density of sphere, rS surface-charge
density of surface, c surface energy between sphere and surface.
Picture and legend adapted with permission from Heinz and Hoh
(1999)
3 Theoretical descriptions
The forces that determine the behaviour of any system
are, ultimately, always the forces between the individual
atoms. Ultimately, every system will, therefore, be best
described by a model that accounts for all the individual
atomic interactions. In practice, this is, however, still out
of reach due to the available computational power limiting system size to, say, 10 nm. Luckily, above this
253
Table 1 Overview of dimensionless numbers and their relation to
the dimension length (at constant velocity); b thermal expansion
coecient (1/K), c surface tension (N/m), g dynamic viscosity
Group
Abbreviation
Bond number
Bo
Grashof number
GrL
Reynolds number
ReL
Weber number
We
Capillary number
Ca
Denition
2
gql qv l
c
gbq2 Ts T1 l3
g2
qvl
g
qv2 l
c
vg
c
Interpretation
Order length
scale, systems can very often be described with continuum theory, which statistically averages the single
interactions. As mentioned in the preceding section,
deviations from the predictions of classical continuum
theory have been observed for both the static and dynamic behaviour of a liquid conned in a space smaller
than 10 molecular diameters, where molecular layers
start being quantised. Molecular dynamics (MD) simulations are, therefore, appropriate for such systems and
can successfully be applied to explain such deviations
(Israelachvili 1992b; Schoen et al. 1989). Recently, MD
simulations were, for example, used to model both
electro-osmotic and Poiseuille ow in small diameter (2nm radius down to 0.95-nm radius) nanochannels (Lyklema et al. 1998; Thompson 2003; Qiao and Aluru
2003). One interesting outcome predicted a reversal of
the direction of the electro-osmotic ow for 3.5-nm wide
slit channels (Qiao and Aluru 2004). MD simulations
have also been employed for the description of liquid
ow in even smaller 0.24 nm inner diameter (6, 6) carbon
nanotubes (Kalra et al. 2003) and in the aquaporins that
conduct water through cell walls (Tajkhorshid et al.
2002). Figure 3, for example, shows a molecular
dynamics simulation of water molecule transport in
aquaporins, illustrating the reversal of the orientation of
the water molecules halfway down the single le they
form inside the channel. This reversal in orientation is
thought to prevent proton conduction by the Grotthus
mechanism (Tajkhorshid et al. 2002). Single-le motion
of water is predicted by MD simulations of carbon nanotubes, aquaporins and also for nanochannels naturally occurring in zeolites (Demontis et al. 2004).
In general, the molecular-scale visualisations that we
have of nanoscale systems, and that we also employ in
some gures in this paper, arise from MD simulations.
MD calculations can also be used for larger systems,
however. In particular, recent MD calculations (with
modern computing power, up to 100 million atoms can
be considered) have been shown to be capable of
describing phenomena induced by thermal uctuations
related to RaleighTaylor instabilities that may provide
insight in turbulent behaviour as well as nanoscale heat
transfer (Kadau et al. 2004).
One length scale larger, we have systems between 10
nm and 100 nm. Smooth-walled liquid-lled channels of
simple geometry and greater than 2 nm in diameter can
254
4 Phenomena
As a result of the palette of forces that act on the
nanoscale, a whole range of phenomena specically occurs on this scale. As already mentioned in the intro-
Fig. 4 Optical micrograph of a water plug (dark colour) in a 108nm high thin-roofed nanochannel. The strange meniscus shape is
due to deection of the channel roof by the large negative pressure
(17 bar) in the water plug
255
256
Fig. 5ac The Stenocara beetle (a), the wax-free condensing areas
on its wings (b) and waxed superhydrophobic wing areas (c). Scale
bars: (a) 10 mm, (b) 0.2 mm, (c) 10 lm. Reprinted with permission
from Parker and Lawrence (2001)
257
258
5 Applications of nanofluidics
A number of applications of nanouidics have already
been mentioned in the previous sections, like the use of
charged polymers for lubrication, the lotus eect for
self-cleaning surfaces, membranes for ltering on size or
charge (e.g. for desalination) and nanoporous materials
for size exclusion chromatography. Furthermore, we
mentioned a number of exemplary natural phenomena,
like (again) the lotus eect for tidy plants and thirsty
beetles, biolubrication, snapping shrimps, passive selective transport in aquaporins and active transport in ion
channels, molecular motors like kinesin and chargebased ltration in the kidney basal membrane. The old
adage natura artis magistra in this respect certainly
applies to nanouidics. In this section, we will mention
some more applications of modern nanouidics and
discuss some ongoing investigations that might lead to
new applications.
5.1 Separation science: size fractionation in regularly
structured microarrays
259
Fig. 10 The hydrogen-bonded
tubular architecture of the selfassembled eight-residue cyclic
D,L-a-peptides (top), and modes
of membrane permeation
accessible to peptide nanotubes
(bottom). Reprinted with
permission from FernandezLopez et al. (2001)
260
Fig. 11 a Schematic of the
behaviour of short and long
pieces of DNA. The short DNA
can follow the sum vector of
both the strong and weak elds,
but the long DNA can only
follow the vector of the large
eld since it gets caught behind
the pillars; bd uorescence
micrographs of DNA
separations under dierent eld
strengths and continuous ow;
d all four DNA lengths (61, 114,
158 and 209 kb) are collected in
dierent channels. Reprinted
with permission from Huang
et al. (2002)
applications of nanoscale rectication ratchets were already mentioned, namely, the asymmetric nanopores for
ion (Siwy and Fulinski 2002) and liquid (Hod and Rabbani 2003) transport. It is to be expected that exciting
devices can be produced when more nanoscale ratchet
devices are produced, since nature employs the concept as
well for transport and pumping, rectifying the abundant
kT noise at the molecular level (Magnasco 1993).
5.3 Separation science: entropic principles
It is energetically favourable for a DNA molecule to be
in the coiled state since this maximises its entropy
261
(Bustamante et al. 1994). Two separation devices employ this tendency, namely, the entropic trap (Han et al.
1999) and the entropic recoil device (Turner et al. 2002).
The entropic trap device (see Fig. 15) uses a sequence of
micrometre-deep channel regions and 90-nm deep
channel regions.
When DNA is transported through the device by
electrophoresis, it is caught in the entropy wells of the
deep regions and only moves through if, by thermal
movement, a sucient part of its chain moves into the
shallow region. Since the chance of this happening is
bigger for longer DNA molecules, these move faster
through the array. The entropic recoil device (see
Fig. 16) employs a channel with alternating densely
pillared and open areas, along which, intermittently, an
electrical eld is applied. Only the DNA molecules that,
during the eld pulse, have entirely moved into the pillared area will remain there; the other molecules, by
recoil, will move back to the original empty channel
section. As a result, short DNA moves faster than long
DNA.
5.4 Single molecule studies: the benets of connement
As already mentioned, much eort has been devoted to
studying the properties of single DNA molecules,
including the determination of the base sequence, and
262
Fig. 16a, b The entropic recoil
device. a DNA molecules are
driven into the pillared area by
the applied electrical eld. b At
zero applied eld, the DNA
molecules recoil except for the
leftmost one, which is entirely
surrounded by pillars.
Reprinted with permission from
Turner et al. (2002)
263
Fig. 18 Schematic of a metallic nanopore or zero-mode waveguide containing an isolated DNA polymerase enzyme. The
engineered nanostructure isolates the uorescent excitation to a
nanometre-scale volume surrounding the enzyme. Transient uorescent bursts represent incorporation events of uorescently
labelled individual bases. Reprinted with permission from Levene
et al. (2003)
possible solution to meet the former objective, at present, oering a resolution of about 50 nm (Richards
2003). This method is well suited for integration in
nanouidic systems, as has already been demonstrated
(Tegenfeldt et al. 2001; Levene et al. 2003). It has been
mentioned earlier that conductometric detection is close
to meeting the former objective, allowing single base
detection during DNA transport through a nanopore
(Deamer and Branton 2002). Many other detection
methods might oer sensitive detection on the nanoscale, but are outside the scope of this paper. Some reviews on detection in microuidic devices will be useful
for the nanouidic regime as well (Vilkner et al. 2004;
Reyes et al. 2002; Auroux et al. 2002; Schwarz and
Hauser 2001).
6.2 Fouling and stability
In nanochannels, molecules will always be very close to
the wall and adsorption of proteins, especially the
amphiphilic proteins, will easily occur, as described by
the DLVO theory. A layer of polyethyleneglycol (also
called polyethyleneoxide) is generally recommended to
avoid adsorption (Tegenfeldt et al. 2004b) and can be
vapour-deposited (Popat and Desai 2004). The working
mechanism is thought to be entropic stabilization of the
PEG coils (Alexanderde Gennes theory) as well as the
stabilisation derived from their excellent hydration
(Morra 2000). An issue that will be encountered in twophase ow when proteins are present is the denaturation
at the liquidwater interface (Clarkson et al. 1999). Also,
high shear rates have to be avoided to maintain protein
264
7 Concluding remarks
The eld of nanouidics has very broad roots in many
disciplines, and the phenomena that are encountered
touch many aspects of physics, chemistry and biology.
The applications of nanouidics presented in this paper
mainly concerned separation science and we saw that, in
the past few years, several fundamentally new modes of
separation were accessed. Two fundamental reasons
seem to lie at the basis of this: rst, we have now entered
the domain where the analyte size is on the order of
magnitude of the device features. Second, absolutely
regular structures can now be manufactured, which
repetitively perform exactly the same operation on the
particles to be separated. It has already been possible to
demonstrate the rst point by the devices for DNA
fractionation because micromachining was sucient for
the separation of (micrometre-sized) large DNA molecules. The second reason actually makes material what
always was the conceptual basis of classical separation
science, namely, that a continuous separation consists of
a sequence of single separation steps (plates). In a
number of nanouidic separation devices, we can now
actually explicitly identify this sequence of plates in time
or space. The precise control over time and space oered
by nanomachining, therefore, holds great promise for
the battle against band broadening.
A second area of nanouidic applications has recently been the study of fundamental properties of liquids and molecules, e.g. in biophysics and uid
mechanics. Here, nanouidics oers the possibility to
conne molecules to very small spaces, or to subject
them to controlled forces. In these areas, nanouidics is
very much used as an engineering tool to extract fundamental knowledge.
What about the future? We will try to identify a few
phenomena and forces that are still untapped. An
important phenomenon that has been hardly used is
specic interaction as occurring, for example, with
antibodies or ion pumps. The challenge here is to devise
structures that harness these specic molecular interactions, if possible, on a continuous and highly parallel
scale. The body does this in a continuous fashion, e.g. in
its molecular machines, and in a highly parallel fashion,
e.g. in muscle contraction. Will this be possible by
technical means as well? Then, this could, for example,
contribute to tissue engineering, but also to actuation or
separation in microsystems or nanosystems. As another
example of an untapped phenomenon with possible
analytical applications, will it be possible to manufacture structures that molecularly exclude protons as
aquaporins do or ions as ion channels do? Can we employ these in nanouidic systems in a controlled way for
separations? A totally dierent and even more speculative question is whether it will be possible to apply
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