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Papers: Part A

2016

Fundamentals and applications of inertial

microfluidics: A review
Jun Zhang
University of Wollongong, [email protected]

Sheng Yan
University of Wollongong, [email protected]

Dan Yuan
University of Wollongong, [email protected]

Gursel Alici
University of Wollongong, [email protected]

Nam-Trung Nguyen
Griffith University, [email protected]

See next page for additional authors

Publication Details
Zhang, J., Yan, S., Yuan, D., Alici, G., Nguyen, N., Ebrahimi Warkiani, M. & Li, W. (2016). Fundamentals and applications of inertial
microfluidics: A review. Lab on a Chip: miniaturisation for chemistry, physics, biology, materials science and bioengineering, 16 (1),
10-34.

Research Online is the open access institutional repository for the University of Wollongong. For further information contact the UOW Library:
[email protected]
Fundamentals and applications of inertial microfluidics: A review
Abstract
In the last decade, inertial microfluidics has attracted significant attention and a wide variety of channel
designs that focus, concentrate and separate particles and fluids have been demonstrated. In contrast to
conventional microfluidic technologies, where fluid inertia is negligible and flow remains almost within the
Stokes flow region with very low Reynolds number (Re < 1), inertial microfluidics works in the intermediate
Reynolds number range (~1 < Re < ~100) between Stokes and turbulent regimes. In this intermediate range,
both inertia and fluid viscosity are finite and bring about several intriguing effects that form the basis of
inertial microfluidics including (i) inertial migration and (ii) secondary flow. Due to the superior features of
high-throughput, simplicity, precise manipulation and low cost, inertial microfluidics is a very promising
candidate for cellular sample processing, especially for samples with low abundant targets. In this review, we
first discuss the fundamental kinematics of particles in microchannels to familiarise readers with the
mechanisms and underlying physics in inertial microfluidic systems. We then present a comprehensive review
of recent developments and key applications of inertial microfluidic systems according to their microchannel
structures. Finally, we discuss the perspective of employing fluid inertia in microfluidics for particle
manipulation. Due to the superior benefits of inertial microfluidics, this promising technology will still be an
attractive topic in the near future, with more novel designs and further applications in biology, medicine and
industry on the horizon.

Keywords
review, inertial, fundamentals, microfluidics, applications

Disciplines
Engineering | Science and Technology Studies

Publication Details
Zhang, J., Yan, S., Yuan, D., Alici, G., Nguyen, N., Ebrahimi Warkiani, M. & Li, W. (2016). Fundamentals and
applications of inertial microfluidics: A review. Lab on a Chip: miniaturisation for chemistry, physics, biology,
materials science and bioengineering, 16 (1), 10-34.

Authors
Jun Zhang, Sheng Yan, Dan Yuan, Gursel Alici, Nam-Trung Nguyen, Majid Ebrahimi Warkiani, and Weihua Li

This journal article is available at Research Online: http://ro.uow.edu.au/eispapers/4850

 Fundamentals and Applications of Inertial Microfluidics: A Review
Jun Zhanga, Sheng Yana, Dan Yuana, Gursel Alicia, Nam-Trung Nguyenb*, Majid Ebrahimi
Warkiani c, Weihua Lia*
a
School of Mechanical, Materials and Mechatronic Engineering, University of Wollongong,
Wollongong, NSW 2522, Australia
b
Queensland Micro- and Nanotechnology Centre, Griffith University, Brisbane QLD 4111,
Australia
c
School of Mechanical and Manufacturing Engineering, Australian Centre for NanoMedicine,
University of New South Wales, Sydney, NSW 2052, Australia

ABSTRACT:

In the last decade, inertial microfluidics has attracted significant attention and a wide variety
of channel designs that focus, concentrate and separate particles and fluids have been
demonstrated. In contrast to conventional microfluidic technologies, where fluid inertia is
negligible and flow remains almost within Stokes flow region with very low Reynolds
number Re <<1, inertial microfluidics works in the intermediate Reynolds number range (~1
< Re < ~100) between Stokes and turbulent regimes. In this intermediate range, both inertia
and fluid viscosity are finite, and bring about several intriguing effects that form the basis of
inertial microfluidics including: (i) inertial migration and (ii) secondary flow. Due to the
superior features of high-throughput, simplicity, precise manipulation and low cost, inertial
microfluidics is a very promising candidate for cellular sample processing, especially for
sample with low abundant targets. In this review, we first discuss the fundamental kinematics
of particles in microchannels to familiarise readers with mechanisms and underlying physics
in inertial microfluidic systems. We then present a comprehensive review of recent
developments and key applications of inertial microfluidic systems according to their
microchannel structures. Finally, we discuss the perspective of employing fluid inertia in
microfluidics for particle manipulation. Due to the superior benefits of inertial microfluidics,
this promising technology will still be an attractive topic in the near future, with more novel
designs and further applications in biology, medicine and industry on the horizon.

1 INTRODUCTION
Microfluidics is referred to the science that deals with the behaviour, precise control and
manipulation of fluids and particles in the scale of tens to hundreds of micrometres 1.
Microfluidics is also considered as a platform for biomedical and chemical applications,
called Lab-on-a-Chip (LOC), or micro Total Analysis Systems (μTAS) 2. Benefiting from the
precise control and manipulation of biological particles and its surrounding
microenvironment, this fascinating technology holds superior advantages compared to
conventional macro-scale platforms (e.g. centrifuge, and flow cytometry etc.). Microfluidics
1
benefits include but is not limited to (i) reduced sample and reagent volumes, (ii) fast sample
processing, (iii) high sensitivity, (iv) low cost, (v) improved portability, and (vi) the potential
to be highly integrated and automated to reduce human intervention and error 3.
Precise manipulation such as focusing, separation and fractionation of bio-particles is an
indispensable capability of microfluidics. For instance, three-dimensional focusing and
ordering of micro-particles along a single line allow for single-cell level detection and
analysis, which is an important component of on-chip flow cytometry. Furthermore,
separation of micro-particles/cells according to their unique biophysical signatures such as
size, density, shape, deformability, and surface proteins allows for a wide range of
applications such as preparation of biological sample (e.g., blood, lymphatic tissue and tissue
digestates etc.), isolation and enrichment of target cells as well as water purification 4, 5.
A number of technologies have already been proposed and developed to manipulate particles
in microfluidic systems. According to the source of the manipulating forces, these
technologies can be categorised as active and passive types. Active technologies such as
dielectrophoresis (DEP) 6, magnetophoresis (MP) 7, acoustophoresis (AP) 8 and optical
tweezer 9 rely on external force fields, whereas passive technologies depend entirely on the
channel geometry or intrinsic hydrodynamic forces, such as pinched flow fractionation (PFF)
10
, deterministic lateral displacement (DLD) 11 and inertial microfluidics 4. An active
technology generally allows for a more precise control of target particles, as well as being
tuneable in real time. However, the flow speed is always limited because the external force
field should overcome the hydrodynamic drag to achieve effective functionality. In contrast, a
passive microfluidic technology is always very simple, robust and works with a relatively
higher flow rate.
Among passive microfluidic technologies, inertial microfluidics has recently attracted a great
attention due to its precise manipulation, simple structure and high throughput. In contrast to
the common sense that, in microfluidics, fluid inertia is negligible (Stokes flow regime, Re →
0, Re=ρUH/µ, where ρ, U and µ are fluid density, average velocity and dynamic viscosity
respectively, and H is characteristic channel dimension), inertial microfluidics works in an
intermediate range (~1 < Re < ~100) between Stokes regime and turbulent regime (Re ~
2000), where both inertia and viscosity of the fluid are finite. The finite inertia of the fluid
brings about several intriguing inertial effects that form the basis of inertial microfluidics
including: (i) inertial migration and (ii) secondary flow.
Inertial migration is a phenomenon where randomly dispersed particles in the entrance of a
straight channel migrate laterally to several equilibrium positions after a long enough distance,
which was first observed more than 50 years ago 12, 13. After the first observation of inertial
migration, a number of experimental studies and theoretical analyses were conducted to
explore the underlying mechanism of this phenomenon 14-21. This phenomenon had not found
its practical application until the emergence of microfluidic technology, where particle size is
comparable with characteristic dimension of microchannel. The inertial migration
phenomenon has been widely recognised by the counteraction of two inertial effects: 1) the
shear gradient lift force FLS, due to the curvature of the fluid velocity profile and its
interaction with a particle, which directs particles away from channel centre, and 2) the wall
2
lift force FLW, a result of the flow field interaction between the suspending particle and the
adjacent walls, which repels the particle away from the wall. The inertial equilibrium
positions of particles result from the balance between these two inertial lift forces, and they
are normally around 0.6 times of channel radius to the channel centreline in a circular channel
13, 22
.
The secondary flow usually appears in a curved channel 23 or a straight channel with
disturbance obstacles 24-26. In a curved channel, the secondary flow is induced by a pressure
gradient in the radial direction, because of fluid momentum mismatch in the centre and near-
wall region within the curvature 4. The fluid elements near the channel centreline have a
higher momentum than that near the wall, therefore flow outwards, and drive relative
stagnant fluid elements near the channel wall inwards along the circumference, forming two
counter-rotating streams, called Dean vortex. Introducing this secondary flow in inertial
focusing brings several benefits. For example, Dean vortex can modify the inertial
equilibrium positions by imposing an additional viscous drag force on particles perpendicular
to the main stream. Size-dependent differential focusing of particles according to the ratio of
inertial lift force and secondary flow drag (FL/FD) promises the complete particle separation.
In addition, Dean vortex could reduce the channel length/footprint, due to the mixing effects
of secondary flow, assisting particles to reach equilibrium position more quickly 4.
In this review, we will first discuss the fundamental kinematics of micro-particles inside
microchannels. We then present a comprehensive review of the recent progress of inertial
microfluidics according to the structure of functional microchannels, because the structure of
microchannel is the most crucial parameter that determines the functionality and performance
of inertial microfluidic devices. Finally, the limitation of current investigations will be
discussed. We then propose several future directions of inertial microfluidics. We hope that
this review could serve as a comprehensive guide for readers who are new to the area of
inertial microfluidics. For readers with a decent background, we hope that the review can
provide extended understanding of fundamental mechanism, inspire novel device design or
initiate expanded application of inertial microfluidics.

2 FUNDAMENTAL DYNAMICS OF PARTICLE MOVEMENT IN A

MICROFLUIDIC CHANNEL

2.1 Viscous drag force

Drag force arises when an object moves through a fluid or, relatively, when the fluid flows
past an object. The origin of the drag force lies in the need to displace the elements of the
fluid out of the way of the moving object. The drag force on a moving spherical particle can
be expressed as:
𝐹𝑑𝑟𝑎𝑔 = 𝑆 ∗ 𝑓𝑑𝑟𝑎𝑔 = 𝜋𝑎2 𝑓𝑑𝑟𝑎𝑔 ⁄4 (1)
where S is the cross-sectional area of moving particles, and a is the diameter of particle. 𝑓𝑑𝑟𝑎𝑔
is viscous drag coefficient which is always determined by the particle Reynolds number
𝑅𝑒 ′ = 𝑣𝑡 𝜌𝑓 𝑎/µ, here 𝑣𝑡 is relative velocity of fluid to particle 27.

3
The viscous drag coefficient 𝑓𝑑𝑟𝑎𝑔 can be divided into four sections according to the range of
particle Reynolds number 𝑅𝑒 ′ 27:
(a) When 10−4 < 𝑅𝑒 ′ < 0.2
µ𝑣𝑡
𝑓𝑑𝑟𝑎𝑔 = 12 (2a)
𝑎

Then
𝐹𝑑𝑟𝑎𝑔 = 3𝜋µ𝑎𝑣𝑡 (2b)
This is actually the well-known formula of the Stokes drag, which has been widely used for
analytical analysis because of its simplicity, especially when relative velocity of fluid to
particle 𝑣𝑡 is very small.
(b) If 0.2 < 𝑅𝑒 ′ < 500~1000
µ𝑣𝑡
𝑓𝑑𝑟𝑎𝑔 = 12 (1 + 0.15𝑅𝑒 ′0.687) (3a)
𝑎

then
𝐹𝑑𝑟𝑎𝑔 = 3𝜋µ𝑎𝑣𝑡 (1 + 0.15𝑅𝑒 ′0.687 ) (3b)
(c) If 500~1000 < 𝑅𝑒 ′ < 2 × 105
𝑓𝑑𝑟𝑎𝑔 = 0.22𝜌𝑓 𝑣𝑡2 (4a)
then
𝐹𝑑𝑟𝑎𝑔 = 0.055𝜋𝑎2 𝜌𝑓 𝑣𝑡2 (4b)
Subsequently, Khan and Richardson examined the experimental data and suggested an
equation that suits for values of 𝑅𝑒 ′ up to 105 27:
𝑓𝑑𝑟𝑎𝑔 = (1.84𝑅𝑒 ′−0.31 +0.293𝑅𝑒 ′0.06 )3.45 𝜌𝑓 𝑣𝑡2 (5a)
𝜋
𝐹𝑑𝑟𝑎𝑔 = 4 𝑎2 𝜌𝑓 𝑣𝑡2 (1.84𝑅𝑒 ′−0.31 + 0.293𝑅𝑒 ′0.06 )3.45 (5b)

In inertial microfluidics, viscous drag force is comprised of two components: along 1)

mainstream direction, due to the axial velocity difference between fluid and the suspended
particles; 2) cross-section, due to the secondary flow induced by the channel curvature, or
disturbance structures.

2.2 Diffusion
Brownian motion is the random motion of suspending particles immersed in a fluid (liquid or
gas), which is resulted from their frequent collision with the quick atoms or molecules in the
gas or liquid. Diffusion is ubiquitous in particle-fluid system. According to Einstein-
Smoluchowski theory, the mean square distance that a particle diffuses in time t is expressed
as 28:
< 𝑟 2 >= 6𝐷𝑡 (6)

4
where D is the diffusion coefficient. According to the Stokes-Einstein relationship, the
diffusion coefficient is given as:
𝑘𝑇
𝐷 = 3𝜋𝜇𝑎 (7)

Where k is the Boltzmann’s constant, which is about 1.3806488×10-23 J/K. T is the absolute
temperature 28. From the above equation, we can see that the higher the temperature T, the
larger is the diffusion coefficient D. In contrast, the larger the particle size a or the higher the
medium viscosity µ, the smaller is the diffusion coefficient. When the particle-fluid system is
not static, but moving along the microchannel, the Péclet number can be employed to
evaluate the relative importance of convection to diffusion 29:
𝐿𝑈
𝑃𝑒 = (8)
𝐷

where L is characteristic dimension of channel and U is characteristic velocity of fluid. The

large Péclet number indicates that convection dominates the transport of particles in the flow,
while a small Péclet number means that diffusion plays a primary role in transport. In inertial
microfluidics, diffusion is normally neglected in the analysis of microscale particle
movement in inertial microfluidics.

2.3 Magnus force--rotation-induced lift force

In an in-viscous flow with uniform velocity U, a stationary cylinder is rotating with constant
angular velocity Ω, see figure 1(a). Assuming no slip velocity at the surface of the cylinder,
the fluid velocity at the bottom part is lower than the velocity at the upper part. According to
the Bernoulli principle, the pressure is higher at the lower part than that in the upper part of
the cylinder. As a result, a lift force FLR is developed to lift the cylinder. The magnitude of
this lift force, per unit length of the cylinder is 30:

𝐹⃗𝐿𝑅 = 𝜋𝜌𝑓2 𝑎𝑈 ⃗⃗
⃗⃗ × 𝛺 (9)
Similarly for a rigid sphere rotating in a fluid, a lateral lift force appears due to the transverse
pressure difference, and this force is called “Magnus lift force” 31:
1
𝐹⃗𝐿𝑅 = 8 𝜋𝑎3 𝜌𝑓 𝑈 ⃗⃗
⃗⃗ × 𝛺 (10)

If the sphere is not stationary, but simultaneously translating through the fluid with a velocity
𝑢 ⃗⃗ by relative velocity (𝑢
⃗⃗𝑝 , replacing the fluid velocity vector 𝑈 ⃗⃗𝑓 − 𝑢
⃗⃗𝑝 ) results in:
1
𝐹⃗𝐿𝑅 = 8 𝜋𝑑 3 𝜌𝑓 (𝑢
⃗⃗𝑓 − 𝑢 ⃗⃗
⃗⃗𝑝 ) × 𝛺 (11)

The direction of Magnus force is perpendicular to the plane defined by the vectors of the
relative velocity and the axis of rotation. In the case of a sphere rotating with an angular
velocity ⃗Ω
⃗⃗s in a rotational flow field, the vector ⃗Ω
⃗⃗ represents the relative rotation between the
fluid and the sphere:
⃗Ω
⃗⃗ = ⃗Ω
⃗⃗s − 0.5∇ × 𝑢
⃗⃗𝑓 (12)

5
Magnus force can be considered as a result of the pressure difference induced by the
streamline asymmetry, due to the rotation of the sphere. Although above particular
expressions are only valid for low Reynolds numbers, this kind of lift force due to the
rotation of a body is also present in a more strongly inertial flow 32. In fact, as a result of the
outer velocity field disturbance by the rotation, where fluid inertia is essential, the Magnus
force would not appear in the creeping flow, but present in all rotating flows with finite
relative velocity no matter it is viscous or inviscid 30.

Figure 1 (a) Rotation induced lift force for a rigid cylinder or sphere in a uniform flow. (b) Saffman force on a
sphere due to slip-shear motion in a simple shear flow. The direction of lateral lift force always directs toward
the side where magnitude of relative velocity is maximum. (c) Boundaries retard the motion of the particle no
matter whether the main flow direction is parallel (I) or perpendicular (II) to the wall boundary. An additional
wall lift force directs perpendicularly to the main flow direction when particles are moving parallel to the
boundary. (d) Shear gradient lift force on a particle in a Poiseuille flow. In the moving frame with the particle,

6
the relative velocity of fluid to particle is larger on the wall side due to the parabolic fluid velocity profile.
Therefore, shear-gradient lift force directs toward the near-by wall 33.

2.4 Saffman force--slip-shear induced lift force

Inertial migration is an interesting phenomenon where randomly dispersed particles in the
entrance of a straight micro-channel migrate laterally to a narrow annulus at about 0.6 times
of the tube radii from the axis, which was first quantitatively measured by Segre and
Silberberg in 1960s 12, 13. After that, a series of analytical analysis and numerical simulation
have been conducted to explain this counter-intuitive phenomenon 14-19. However, the full
problem is more complex, as not only the effect of inertia needs to be calculated for a particle
in a parabolic velocity profile, but also the presence of the tube walls must be taken into
account. The walls are clearly very important to the existence of the phenomenon. Without
channel walls, the flow will be uniform and there is even no velocity gradient, generating no
rotational motion on a force-free rigid sphere, and consequently no lateral lift force appears.
The wall effects at least act in two different ways: First, the existence of walls creates a fluid
velocity gradient (shear rate), and corresponding shear-induced particles rotation. The extra
drag caused by walls makes the particle lagging behind the fluid. This slip-shear motion will
generate a lateral force on the particles called the Saffman force. Second, the flow field
around the particle is disturbed by the presence of the walls and the inertial effects will differ
from those for a particle in an unbounded flow, especially when the particle is close to the
walls 34. Here, we focus on the first effect and only consider the force on a sphere in an
unbounded simple shear flow, i.e. constant shear rate and zero shear gradient. The second
effect of the wall will be detailed in the section of “Wall-induced lift force”. For Poiseuille
flow with non-zero shear gradient, the effects of parabolic velocity profile on particle
migration will be discussed in the section of “Shear-gradient lift force”.
Using the matched asymptotic expansion method, Saffman 34 calculated the lateral lift force
on a sphere in a simple unbounded shear flow. The force arises from the interaction of the
Stokeslet velocity field of the particle and the velocity gradient of the bulk flow. The
magnitude of this force is:
𝐾
𝐹𝑆 = 4 𝑉𝑎2 (𝛾𝑣 −1 )1⁄2 (13)

where K is a numerical constant (K~81.2), γ is velocity gradient (or shear rate), υ is the
kinetic viscosity and V is the relative velocity between particle and fluid at the streamline
through the centre of the particle. It should be pointed out that this force is the only effect of
shear rate, independent of the rotation of particle. In the real situation, the spheres are free to
move, and a relative rotation will be induced by the shear. Consequently, a Magnus-type
force may appear on the sphere. In most practical flows, the effects of rotation and shear
cannot be considered as independent, and a simple superposition of the two lift forces would
inevitably bring errors 30. Unless the rotation speed is much greater than the rate of shear, for
1
a freely rotating particle Ω = 2 𝛾, Saffman force 𝐹𝑆 , caused by the interaction of slip velocity
(Stokeslet flow field) and shear, will generally be at least one order of magnitude larger than
Magnus force 𝐹𝐿𝑅 caused by the interaction of slip and particle rotation at low Reynolds
numbers 32.
7
The direction of Saffman force is always towards the side where the magnitude of relative
velocity V is maximum, Figure 1(b). If the particles are leading the flow, Saffman force
directs to the stagnant wall in a simple shear flow; if the particles are lagging the flow,
Saffman force points to the moving wall. In the case of a neutrally buoyant particles in a
Poiseuille flow, the particle is force-free and the Stokeslet generated by the lag of the particle
relative to the shear flow (slip-shear) is balanced by an opposite Stokeslet originating from
the curvature of the base flow, so there is basically no net Stokeslet in the base flow,
consequently no Saffman force acts on the sphere 32. Saffman force is more relevant in the
case of non-neutrally buoyant particles in a vertical flow, or additional outer force (electrical
or magnetic) 35, 36 acting on the particle to lag or lead fluid flow, creating obvious net
Stokeslet flow field. In this situation, the Saffman force will direct to the channel centreline
when particles lag the flow, or direct toward the channel walls when particles lead the flow.

2.5 Wall-induced lift force

In the previous section, we mainly talk about the first effect of walls. The existence of wall
creates a velocity gradient (shear rate) of fluid, and possibly causes the rotation of spherical
particles due to the shear. Thus, transverse Magnus force and Saffman force may act on the
particles to migrate particles laterally. In addition, the second effect of walls is the change of
the flow field around a particle in the presence of the walls. The net force on a particle near
the wall differs from that of a particle in an unbounded flow. In order to eliminate effects of
shear rate to the lateral lift force, one possible scheme is investigating the motion of particles
near a wall in a stagnant fluid.
In general, the effect of a wall on the motion of immersed objects is to retard the motion of
the object in both parallel and perpendicular directions, as well as exerting a transverse
migration motion. Two distinct interactions between an immersed object and a wall exist: (i)
The motion of the immersed object is primarily influenced by a single wall on one side of the
object. Any other walls are too far from the immersed object to influence significantly its
motion. In this case, the main effect of the wall are decelerating the particle and driving the
immersed object away from the wall, Figure 1(c). It applies to the situation where the
characteristic dimension of immersed particle is much less than the dimension of the flow
channel. (ii) The immersed object is surrounded by the boundaries (walls) of the flow domain.
Thus, the boundaries decelerate significantly the motion of the object. It happens if the
characteristic dimension of the immersed particle is of the order of the dimension of flow
channel, such as small bubbles moving in capillaries, and the surrounding walls will constrain
the motion of immersed particles 30.
First, we discuss the situation where the dimension of particles is much smaller than that of
channel. The particles are primarily influenced by a single wall. The direction of the particle
motion can be perpendicular or parallel to the walls. When the immersed sphere moves
perpendicularly to a boundary, the effect of the boundary will retard the motion of the particle,
which is equivalent to the increase of the drag coefficient. Brenner 37 derived a first-order
correction for the drag coefficient of a small sphere in the creeping flow moving towards a
solid wall.

8
 16𝜇𝑣𝑡
𝑓𝑑 = 𝑓 (14)
𝑎
𝑛(𝑛+1) 2𝑠𝑖𝑛ℎ[(2𝑛+1)𝜃 +(2𝑛+1)sinh⁡(2𝜃 ]
𝑓 = 𝑠𝑖𝑛ℎ𝜃𝑤 ∑∞ 𝑤 𝑤
𝑛=1 (2𝑛−1)(2𝑛+3) × [4𝑠𝑖𝑛ℎ2 [(𝑛+0.5)𝜃 ]−(2𝑛−1)2 𝑠𝑖𝑛ℎ2 (𝜃 − 1] (15)
𝑤 𝑤)

where 𝜃𝑤 = 𝑐𝑜𝑠ℎ−1 (2 𝑙𝑤 ⁄𝑎) is a function of the ratio of distance from sphere centre to the
wall to the particle diameter, 𝑙𝑤 is the distance of sphere centre to the channel wall, Figure
1(c). Equation (15) indicates that the hydrodynamic force on the particle increases
significantly when the particle approaches the wall.
When the immersed sphere moves parallel to the boundary, a transverse lift force repels the
particle away from the wall. Given the dimensionless distance 𝑑 ∗ ≪ 1 , where 𝑑 ∗ =
𝜌𝑓 𝑙𝑤 𝑣𝑠 ⁄𝜇 , the lateral migration velocity can be expressed as 38, 39:
3 11
𝑣𝑧 = 64 𝑅𝑒𝑠 𝑣𝑠 {1 − 32 𝑑 ∗ 2 + ⋯ } (16)

where vs is the sedimentation velocity of the sphere in a quiescent fluid, and 𝑅𝑒𝑠 is a
Reynolds number defined by the sedimentation velocity and particle diameter, expressed as
𝑅𝑒𝑠 = 𝜌𝑓 𝑎𝑣𝑠 ⁄𝜇. Considering the implied condition of 𝑑 ∗ ≪ 1, it can be further simplified as:
3
𝑣𝑧 = 64 𝑅𝑒𝑠 𝑣𝑠 (17)

Equation (17) indicates that the migration velocity of a sphere is constant when it is in the
vicinity of the wall. And this expression was experimentally validated by the data of
Cherukat and McLaughlin 40 up to 𝑅𝑒𝑠 = 3.0.
In addition, when 𝑑∗ ≫ 1, the migration velocity then becomes 39:
3 ⁄2
𝑣𝑧 = 16 𝑅𝑒𝑠 𝑣𝑠 (𝑑 ∗ −2 + 2.21901𝑑 ∗ −5 +⋯) (18)

It indicates that for large values of 𝑑 ∗ (large distance between sphere and wall or large
sedimentation velocity of sphere), the migration velocity decreases continuously and tends to
zero as 𝑑 ∗ → ∞.
In addition to the lateral lift force that repels particle away from the wall, the effect of the
boundary will also retard the sedimentation motion of the particle, corresponding to an
increased drag force. The drag force experienced by a sphere settling down in the
𝑎 1
neighbourhood of a plane wall (2𝑙 ≪ 1 ≪ 𝑅𝑒 ) is 39:
𝑤 𝑠

3 9 𝑎
𝐹𝑑 = 3𝜋𝜇𝑎𝑣𝑠 (1 + 8 𝑅𝑒𝑠 + 32 𝑙 + ⋯ ) (19)
𝑤

𝑎 1
At a relative large distance (2𝑙 ≪ 𝑅𝑒 ≪ 1), the drag force is expressed as 39:
𝑤 𝑠

3 𝑎 1 3⁄2
𝐹𝑑 = 3𝜋𝜇𝑣𝑠 {1 + 8 𝑅𝑒𝑠 − 0.095 𝑙 (𝑅𝑒 ) +⋯} (20)
𝑤 𝑠

In a bounded flow, when the characteristic dimension of the sphere is of the same order of
magnitude as the size of the flow channel, the surrounding boundaries provide a physical
constraint to the flow, and in general, the immersed objects move close to the centreline of

9
the channel. In order to prevent channel from being blocked, the blockage ratio δ which is the
ratio of particle diameter to channel diameter should be less than 1. Besides, the boundaries
decelerate significantly the motion of the object. A wall drag multiplier of rigid sphere
moving in a cylindrical tube was derived and expressed as 30:
𝐹
𝐷
𝐾𝑤𝑎𝑙𝑙 = 3𝜋𝑎𝜇𝑣 = [1 − 2.0144𝛿 + 2.0888𝛿 3 − 6.948𝛿 5 − 1.372𝛿 6 + ⋯ ]−1 (21)
𝑠

In summary, the effects of walls on the motion of immersed spheres in a tube are to
decelerate the motion of the sphere, and repel particles to the centreline of channel.

2.6 Shear gradient lift force

As discussed above, the particle will lag the flow due to the effects of wall. If the curvature of
the undisturbed fluid velocity profile is zero, it becomes a simple shear flow, Figure 1(b).
Then the pressure will be higher on the left, pushing particles to the centreline of channel.
However, in the Poiseuille flow, the Stokeslet generated by the lag of the particle relative to
the shear flow (slip-shear) may be balanced by an opposite Stokeslet originating from the
curvature of the base flow. Furthermore, the curvature of fluid velocity profile can even
reverse this trend. Figure 1(d) clearly shows that the magnitude of relative velocity of fluid to
particle is much higher on the left side of particle than that on the right side, due to the
parabolic nature of velocity profile. And it overwhelms the asymmetry caused by lag velocity.
Similar to Saffman force, the dissymmetry of relative velocity causes a low pressure on the
left/wall side, generating a shear gradient lift force that’s opposite to the Saffman force. The
Shear-gradient lift force leads particles migrating toward walls until the wall lift force repels
and balances it 32, 33.

2.7 Net inertial lift force

For a neutrally buoyant rigid sphere flowing in a straight wall-bounded Poiseuille flow,
besides a viscous drag force along the axis, there are four lateral forces acting on the sphere:
Magnus force due to slip-rotation, Saffman force due to slip-shear, wall lift force due to the
disturbance of flow field around particles from wall, and shear gradient lift force due to the
curvature of undisturbed fluid velocity profile. Among them, Magnus force and Saffman
force are often very small and negligible. Shear gradient lift force, directing particles toward
channel walls, and wall lift force, repulsing particles toward channel centreline, are
commonly recognised as the dominant effects for the lateral migration of the particle 35. The
balance of shear gradient lift force and wall lift force creates several equilibrium positions
halfway between the channel walls and centreline, and this theory can explain the observation
of Segre and Silberberg reasonably 12, 13, Figure 2(a). It should be noted that such a
breakdown is rather unconventional, since an unbounded parabolic velocity profile cannot be
realized in practice, but permits the separation of the effects due to the curvature of the
undisturbed velocity profile and the wall-induced disturbance 17.
Through the method of matched asymptotic expansions, Asmolov 17 derived an analytical
expression of the net inertial lift force acting on a small rigid sphere ( 𝑎/𝐻 ≪ 1) in a
Poiseuille flow:

10
 𝐹𝐿 = 𝑓𝐿 𝜌𝑓 𝛾 2 𝑎4 (22)
The force can be further simplified as:
𝐹𝐿 = 𝑓𝐿 𝜌𝑓 𝑈 2 𝑎4 ⁄𝐻 2 (23)
The hydraulic diameter H is defined as 𝐻 = 𝐷 for a circular channel (D is the diameter of
circular cross-section) or 𝐻 = 2𝑤ℎ⁄(𝑤 + ℎ) for a rectangular channel (w and h correspond
to width and height of the rectangular cross-section). The lift coefficient 𝑓𝐿 is a function of
the particle lateral position x and channel Reynolds number Re 17, 41, Figure 2(b). The lateral
position 𝑥𝑒𝑞 where 𝑓𝐿 = 0 corresponds to the inertial equilibrium position. It should be noted
that the channel centreline 𝑥 = 0 is not a stable equilibrium position, since a little deflection
from centreline will never return the particles back. It also has been found that 𝑓𝐿 decreases
with increasing Re (or 𝑈𝑓 ), suggesting that inertial lift force scales less strongly than ⁡𝑈𝑓2 4.
Recently, Zhou and Papautsky 42 derived a scaling for the lift coefficient based on their
𝐻2
experimental results: 𝑓𝐿 ∝ 𝑎2 .. Although fL varies with Reynolds number, at Re<100 that
√𝑅𝑒
is typical for most microfluidic applications, the lift coefficient remains relatively constant,
and can be approximated averagely as 𝑓𝐿 = 0.5 43.
For a finite-size particle (0.05 ≤ 𝑎⁄𝐻 ≤ 0.2), particle would cause disturbance to the main
channel flow. Di Carlo et al. 44 calculated the inertial lift forces through finite element
simulation, taking into account the finite-size effects of the suspended particles. The net force
was scaled near the channel centre and channel wall respectively. Near the channel centre,
where the effects of the wall are weak, the lift force scales as 𝐹𝐿 ∝ 𝜌𝑓 𝑈𝑓2 𝑎4 /𝐻 2 . While near
the channel wall, wall effects dominate, and it scales as 𝐹𝐿 ∝ 𝜌𝑓 𝑈𝑓2 𝑎6 /𝐻 4 .

11
Figure 2 (a) Balance of shear gradient lift force and wall lift force results in the inertial equilibrium positions in
a Poiseuille flow. (b) The net lift coefficient is a function of particle lateral position x and Reynolds number Re
22
. (c) Deformability-induced lift force directs to the channel centreline, and shifts inertial equilibrium positions
closer to the channel centre, compared with the rigid spherical particles 45.

2.8 Deformability-induced lift force

Although solid rigid particles can be used as a simple model in the study of hydrodynamic
behaviour of particles in a microchannel, the practical bio-particles such as cells and vesicles
are not rigid but deformable. The deformability will induce additional lift forces on the
particles. The deformability-induced lift force is perpendicular to the main streamline, and it
is believed to be the effects of shape-change of particle and nonlinearities caused by the
matching of velocities and stresses at the deformable particle interface 35. Three
dimensionless parameters can be used to characterize the relative deformation of a droplet: i)
𝜌𝑓 𝑈 2 𝑎
Weber number, 𝑊𝑒 = (inertial stress vs. surface tension), where σ is the surface
𝜎

12
 𝜇𝑈𝑎
tension; ii) Capillary number, 𝐶𝑎 = (viscous stress vs. surface tension); and iii) Viscosity
𝜎ℎ
ratio, 𝜆𝑑 = 𝜇𝑑 /𝜇, here µd is the dynamic viscosity of fluid inside the droplet 35.
Chan and Leal derived an analytical expression for deformability-induced lift force, given the
conditions that the drop or bubble is not too close to the walls 46, 47:
𝑎 𝑑
𝐹𝐿,𝑑𝑒𝑓𝑜𝑟𝑚𝑎𝑡𝑖𝑜𝑛 = 𝜇𝑈𝑎(𝐻)2 𝐻 𝑓(𝜆𝑑 ) (24)
16𝜋 11𝜆𝑑 +10 3 19𝜆𝑑 +16
𝑓(𝜆𝑑 ) = (𝜆 3
[ (3𝜆3𝑑 − 𝜆𝑑 + 8) + (2𝜆2𝑑 − 𝜆𝑑 − 1)] (25)
𝑑 +1) 140 14 3𝜆𝑑 +2

where d is the distance between drop and centre of channel. Furthermore, the condition of
𝜆𝑑 < 1 or 𝜆𝑑 > 10 was specified for the deformability-induced lift force directing towards
the center of the channel 47.
The experimental measurements of the inertial lift force by Di Carlo et al. 44 was used to
extract an equation for inertial lift force near channel centre where 𝑑 ⁄𝐻 < 0.1. The negative
𝑐𝑒𝑛𝑡𝑒𝑟
sign indicates that this force, 𝐹𝐿,𝑖𝑛𝑒𝑟𝑡𝑖𝑎𝑙 , is directed towards the walls.
𝑐𝑒𝑛𝑡𝑒𝑟 𝑑
𝐹𝐿,𝑖𝑛𝑒𝑟𝑡𝑖𝑎𝑙 = −10𝑅𝑝 𝜇𝑈𝑎 (𝐻) (26)

When both drop deformation and inertial lift forces are small (for 𝐶𝑎 < 0.01⁡𝑎𝑛𝑑⁡𝑅𝑝 <
0.01), Stan et al. 46 introduced an empirical formula for net lift force on drops or bubbles as:
𝑎 3 𝑑
𝐹𝐿,𝑒𝑚𝑝𝑖𝑟𝑖𝑐𝑎𝑙 = 𝐶𝐿 𝜇𝑈𝑎 (𝐻) (𝐻) (27)

Here lift coefficient 𝐶𝐿 must be determined from experimental measurements for each pair of
the continuous and dispersed phases. And 𝐶𝐿 ~31.25 can be used for the order-of-magnitude
estimate 46.
Deformability-induced lift force can be used to separate and enrich malaria-infected red
blood cells (iRBCs) from normal healthy RBCs (hRBCs) for the diagnosis of malaria. The
parasite releases proteins that trigger the cross-linking of the spectrin network in the iRBC’s
phospholipid bilayer membrane, thus increases the rigidity of the iRBCs. Normal hRBCs are
more deformable than iRBCs, migrating towards to the centre of the channel. Due to the
massive hydrodynamic interactions and mechanical collisions between the RBCs in high
haematocrit (Hct) blood, stiffer malaria iRBCs are displaced towards the sidewalls, and can
be depleted and enriched by bifurcated outlets 48.
In inertial microfluidics, Hur et al. 45 found that centre-directed deformability-induced lift
force shifts inertial equilibrium positions a little closer to the channel centreline than that of
rigid particles, Figure 2(c). By the combination of size and deformability, circulating tumor
cells (CTCs) with more deformability than the cells from the same organ, has been
demonstrated to be separated and enriched from peripheral blood 45.
Besides particle deformability, the shape of particles 49 and the properties of medium 50 also
impact the inertial migration and equilibrium positions, which will not be discussed here. A
summary of particle kinetics in inertial microfluidics is shown in Figure 3.

13
Figure 3 Outline of particle kinetics in inertial microfluidics. There are four aspects impacting inertial focusing
in inertial microfluidic systems: channel walls, channel curvature or disturbance structure, particle and medium
property. Channel walls create wall lift force, shear gradient lift force, slip-shear lift force and rotation lift force
on particles, while front two lift forces are widely recognised as responsible for inertial migration phenomenon.
Channel curvature or disturbance structure generates secondary flow, which assists inertial focusing process and
modifies final inertial focusing positions. Besides, the properties of particle (e.g. size, shape and deformability)
and medium (Newtonian, Viscoelastic) also influence the final inertial focusing positions.

3 THE RECENT PROGRESS OF INERTIAL MICROFLUIDICS

According to the structure of the functional microchannel, the reported inertial microfluidic
devices can be categorised according to their channel structure as: (i) straight channels, (ii)
14
spiral channels, (iii) straight channels with pillar arrays or expansion-contraction arrays, and
(iv) serpentine channels. In this chapter, we review the current progress of inertial
microfluidics, including the phenomenon and possible explanation of inertial migration in
different structured micro-channels, as well as their application in biomedicine and disease
diagnosis. The characteristics of inertial forces in different structured channels were
summarized in Supplementary Table 1.

3.1 Straight channel

In a straight channel with a circular cross-section, the randomly distributed particles migrate
laterally to a narrow annulus at about 0.6 times of the channel radii form the axis 12, 22, as
depicted in Figure 4(a). However, in a straight channel with a rectangular cross-section,
which is the most widely used structure due to the limitation of microfabrication, the situation
becomes more complex. In a square straight channel (AR=height/width=1), particles normally
focus to four equilibrium positions 51, facing the centre of each wall, Figure 4(b).
Additionally, a further reduction to two equilibrium positions happens in a low aspect ratio
(AR ≈ 0.5) channel, particles focus at the centre about 0.2H away from the long walls (see
Figure 4(c)) 42, 52, 53. One explanation about this equilibrium positions’ reduction was given
by Zhou and Papautsky 42 by a two-stage migration model which is illustrated in Figure 4(c).
In a straight channel, particles’ lateral migration velocity (UL) and the minimum channel
length (Lmin), which is the minimum length that is required for particles to migrate to their
inertial equilibrium positions, can be derived by balancing the net inertial lift force with
Stokes drag (see Figure 4(d)) 41:
Fstks  3aU L (28)

FL  f U 2a3
UL   (29)
3a 6H 2

H 3 f H 3
Lmin  U  (30)
2U L  f Ua 3

Two dimensionless Reynolds numbers can characterize the lateral migration of particles in
straight channel: channel Reynolds number Re describing the ratio between inertial force and
viscous force of fluid in a flow, and particle Reynolds number RP additionally considering the
size ratio of particle to channel.

a2  f Ua 2
R P  Re  (31)
H2 H

When RP<<1, particles are subjected to the dominant viscous drag to follow fluid streamlines.
However, increasing RP to the order of 1, inertial lift forces become dominant and lateral
migration of particles across the fluid streamlines becomes obvious 43.

15
Figure 4 Inertial equilibrium positions in a straight channel with different cross sections: (a) Circular cross
section; (b) square cross-section, and (c) rectangular cross section with low aspect ratio (≈ 0.5). Illustration of
two-stage migration model proposed by Zhou and Papautsky 42. (d) The lateral migration speed UL and
minimum channel length for particle focusing Lmin.

Although straight channel is often employed as a basic model to investigate the fundamental
mechanism of inertial migration due to its simplicity 18, 42, 44, 54-56, this channel type has been
utilized for a variety of applications. Hur et al. 57 demonstrated the purification of adrenal
cortical progenitor from digestions of murine adrenal glands utilizing hydrodynamic inertial
lift forces in a straight micro-channel with high aspect ratio, Figure 5(a). The inertial focusing
position depends on the cell size, where larger cells are closer to the channel center, and
smaller cells are closer to the channel walls. Differentially focused cells can be collected at
designated outlets based on apparent diameter.

16
Figure 5 (a) Label free isolation of adrenal cortical progenitor cells by size-based differential inertial focusing in
a straight channel. Reproduced from Ref. 57, an open-access article. (b) Separation of pathogenic bacteria from
diluted blood in a series-connected straight channel which utilizes a unique differential transit time by size-
dependent inertial lift forces. Reproduced from Ref. 58 with permission from John Wiley and Sons. (c) Complete
separation of particles in a cascaded channel with two straight segments with different aspect ratios (ARs).
Reproduced from Ref. 59 with permission from the Royal Society of Chemistry. (d) Rapid inertial solution
exchange in a straight channel for controlled interaction of reagents with cells and particles. Reproduced from
Ref. 60 with permission from American Chemical Society Copyright (2014).

Mach and Di Carlo 58 reported a massively parallelized microfluidic device that passively
separates pathogenic bacteria from the diluted blood (Figure 5(b)). The device consists of 40
single straight micro-channels placed as a radial array. Each channel consists of three
segments with different cross-sections, which uses a unique differential transit time by size-
dependent inertial lift forces to obtain cell separation. The authors demonstrated that more
17
than 80% pathogenic bacteria can be removed after passing two times of the same system.
The parallel device can process 240 mL/h with an extreme high throughput of 400 million
cells per minute. Later, Zhou and colleagues 59 utilized a modified design on size and length
for their cascaded straight channels, as shown in Figure 5(c). The separation concept is based
on the theory of the two-stage inertial migration, which permits precise prediction of particle
or cell position within the micro-channel. Randomly-distributed particles first flow through a
high AR channel (segment I) where particles are focused at the half of channel height near
two sidewalls. Then the channel expands into a low AR channel (segment III), which
modifies equilibrium positions to the centres of top and bottom walls. Since larger particles
have a much higher migration velocity, they reach the relocated points very quickly. In
contrast, lift forces on smaller particles are too weak to alter their lateral position. This effect
enables a separation of particles by size. After this modification, a much higher separation
efficiency (~ 99%) and purity (~ 90%) were achieved in their work.
Manual concentration, staining and washing procedures for cellular sample are routinely
conducted by cytopathologists as a means of diagnosing malignancies and other diseases.
However, current approaches for chemical treatments of cells and chemical reactions
typically operate on slow time scales (~seconds to minutes), limiting the realm of fast
molecular events. Inertial lateral migration has been subtly employed as a helpful tool to
mediate millisecond reaction time around particles and cells 60, as shown in Figure 5(d). The
main transfer channel is designed with a low aspect ratio, so particles will migrate to the
lateral centre of a channel. The channel has three inlets. Particles suspension is infused from
the two side inlets, while the reagent is pumped in from the central inlet. Lateral migration of
particles from original medium to the middle stream initiates the contact of particles/cells
with chemical reagent, and duration of particles within middle stream mediates the reaction
time. The proposed microfluidic system can perform several functions in the sample
preparation for cytopathology that (i) automates colorimetric staining on-chip, (ii) images
cells in flow, and (iii) provides additional quantitative analyses of captured images to aid
cytopathologists 61. In addition, ordering particles along a specific stream within a simple
straight microchannel could also be integrated with optical detector to form an on-chip flow
cytometer system. In another study, Hur et al. 62 proposed an extremely high-throughput on
chip imaging flow cytometry, consisting of 256 high-aspect parallel straight channels and
high-speed optical camera, promising interrogation rates up to 1 million cells per second.
In general, a straight channel has the advantage of simplicity and ease of operation. Therefore,
the mechanism of inertial migration phenomenon in a straight channel is relatively clear and
has already been studied extensively. However, since 𝐹𝐿 ∝ 𝐻 −2 , the sizes of channel cross-
section are normally restricted to provide enough lateral lift forces. In addition, the channel is
relatively long, which would add up the flow resistance and lead to a large device footprint.
In the following section, we will see that, the introduction of a secondary flow by channel
curvature or obstacle structure will not only aid the inertial migration progress, but also
modify the final equilibrium positions. However, its mechanism becomes more complex and
demands more rigorous explanation.

18
 3.2 Spiral channel
Introduction of channel curvature along a single constant direction forms a spiral channel.
When fluid flows through a curved channel, a secondary flow arises due to the velocity
mismatch in the downstream direction between fluid in the central and near-wall regions. The
fluid elements near the channel centreline have larger inertia and would tend to flow outward
around a curve due to the centrifugal effect, creating a pressure gradient in the radial direction
within the channel. In a fully-bounded channel, due to the centrifugal pressure gradient,
relatively stagnant fluid near the walls re-circulates inward, finally forming two symmetric
circulating vortices 4, Figure 6(a). The parameters that influence the distribution and strength
of the secondary flow in a curved channel include Dean number K, Reynolds number Re and
aspect ratio of the channel AR. Dean number K is a function of Reynolds number Re, the
hydraulic diameter for the rectangular channel H and the radius of curvature R 23:
𝐾 = (𝐻 ⁄2𝑅 )1⁄2 𝑅𝑒 (32)
Following Squires and Quake 63, the secondary flow velocity scales as
𝑈𝐷 ~𝐾 2 𝜇/(𝜌𝐻) (33)
Furthermore, the magnitude of secondary flow UD can be approximated as 64, 65:

U D  1.8  104  K 1.63 (34)

In addition, the ratio of channel dimension to radius of curvature 𝛿 = 𝐻 ⁄2𝑅 , Dean number K
as well as the aspect ratio AR have important effects on secondary flow distribution 23.
Particles flowing in a curved channel with finite inertia will experience both inertial lift
forces and secondary flow drag. In most cases, the density of particle is very close to that of
fluid, so the effect of particle centrifugal force around channel curvatures is negligible. When
the channel curvature is along a single direction, the curved channel is a spiral channel. And
the direction of secondary flow within each cross-section is constant, although there may be a
little variation on its magnitude due to the change of channel curvature. By a first order
approximation, one can make an assumption that the effects of inertial migration and
secondary flow act in superposition on particles in spiral channels, Figure 6(b). A particle
held stationary at inertial equilibrium positions experiences a secondary flow drag whose
magnitude is directly proportional to the local secondary flow velocity 4. This secondary flow
drag force acts to entrain particles within the streamline of symmetrically rotating vortices,
which is primarily an effect of mixing. In contrast, inertial lift forces tend to hold particles at
specific equilibrium positions within channel cross-section. Therefore, the order of magnitude
scaling between inertial lift forces and Dean drag 𝑅𝑓 = 𝑎3 𝑅 ⁄𝐻 3 determines the final
behaviour of the suspended particles. This dimensionless parameter is useful for the
prediction of particle behaviour in curved channels.
At limiting conditions where (i) 𝑅𝑓 → 0 , Dean drag force dominates the behaviour of
particles, particle streams neglect inertial equilibrium positions, and remain entrained within
the secondary flow streamlines; and (ii) 𝑅𝑓 → ∞, inertial lift force is dominant, particles
migrate to the inertial equilibrium positions independent of the secondary flow 4. For most

19
cases, in the intermediate range of Rf , inertial equilibrium positions will be modified by the
secondary flow, Figure 6(b), leading to very intriguing new focusing phenomenon. It has
been noted that 𝑅𝑓 is dependent on particle size, so that two different-sized particles may
experience different forces in the same curved channel. Therefore, it would be possible to
separate particles by size in spiral channels.
Yoon et al. 66 proposed another kind of particle separation in a curved channel. In their work,
the separation was achieved by the net effect of secondary flow velocity distribution, rather
than the theory of the balance between secondary flow drag and inertial lift forces. Larger
particles with diameter larger than 0.72h experience a net secondary flow drag directing
outward to the channel curvature, while smaller particles with diameter less than 0.27h
experience a net secondary flow drag inward, Figure 6(c) and (d). Therefore, particles can be
separated based on altered net direction of secondary flow drag on different-sized particles.

Figure 6 (a) Dean flow with two counter-rotating vortices are created in curved channels. Reproduced from Ref.
4
with permission from the Royal Society of Chemistry. (b) Superposition of inertial lift force and Dean flow in
a curved channel modifies the number and position of the inertial equilibrium positions. Reproduced from Ref. 4
with permission from the Royal Society of Chemistry. (c) - (d) Size-based separation by the different directions
of secondary flow drag force exerted by the velocity distribution. Reproduced from Ref. 66 with permission from
the Royal Society of Chemistry.

Spiral channels were investigated extensively for particle ordering and separation by
Papautsky’s group 67, 68, Go’s group 66, Jiang’s group 69, 70 and Han’s group 71, 72, etc. Bhagat
et al. 64 proposed a sheath-less, on-chip flow cytometry system based on the principle of

20
Dean coupled inertial microfluidics, Figure 7(a). The microfluidic flow cytometry system
could provide a throughput of 2,100 particles per second. Furthermore, Kemna et al. 65
reported an integrated microfluidic system that combines cell-ordering in spiral channel with
droplet microfluidic generator. Cells were focused in three dimensional along a single
particle chain and ordered with a precise longitudinal space, Figure 7(b). Single cell
encapsulation was successfully demonstrated downstream with efficiency about 80%. Such a
system is very promising in the applications such as cell-based assays, drug screening and
cell printing technologies.
Passive separation and sorting of particles/cells is one of superior advantages for spiral
microchannels, benefiting from the distinct size dependence characteristics of secondary flow
and inertial lift forces. In another work, Bhagat and colleagues 67 demonstrated a complete
separation of 7.32 µm and 1.9 µm polystyrene particles at Dean number of 0.47 in a 5-loop
spiral microchannel. Later, Kuntaegowdanahalli and co-workers 68 from the same group
extended the separation capability of spiral channels. The authors demonstrated continuous
separation of triplet polystyrene beads (10, 15 and 20 µm in diameter) with an efficiency of
90% and a throughput of 1×106 cells/min in a spiral channel, Figure 7(c). Furthermore, this
separation ability was employed to separate different-staged cells for cell cycle
synchronization, which is essential for studying cellular properties, biological processes and
elucidating genetic regulatory mechanisms and events involved in each phase prior to cell
division 73.
Cancer, also known as a malignant tumour, is a group of diseases involving abnormal cell
growth with the potential to invade or proliferate in other organs of the body. Metastases
from primary tumours are the leading causes of death (~90%) for nonhematological cancers
74
. During the progression of metastasis, cancer cells escaped from solid tumours and enter
the bloodstream, becoming circulating tumour cells (CTCs), which hold a great potential to
serve as important biomarkers for early diagnosis of cancer metastases, as well as cancer
prognosis and therapy monitoring. CTCs analyses are considered as a real-time “liquid
biopsy”, which is much less invasive than the current method for cancer diagnosis requiring
invasive biopsy followed by molecular analysis. However, CTCs are extremely rare,
comprising only a few out of one billion haematological cells in blood, making their isolation
and characterization an extreme technological challenge.
In an intriguing work, Hou et al. 75 employed a spiral channel to isolate CTCs from blood
under the assistance of a sheath flow, and it achieved a recovery rate of more than 85%. This
CTCs isolation platform is working in a label-free and clog-free continuous manner,
significantly reducing the capturing costs and alternation of CTCs’ morphology. In addition,
a clinical validation with positive detection of CTCs from all the patients’ blood samples was
reported. Meanwhile, Sun and colleagues 69, 70 reported a passive double spiral microfluidic
device, Figure 7(d), which can separate and enrich tumour cells (MCF-7 and Hela cells) from
diluted whole blood with a throughput of 3.33×107 cell/min. Compared to the single spiral
microchannel, the double spiral device was claimed to provide better focusing behaviour of
small blood cells and improved separation efficiency. To further scale up the throughput of
spiral channels, parallelization technology must be employed. Warkiani et al. 76 reported a

21
multiplexed microfluidic device stacked by three spiral channels along vertical direction for
ultra-high throughput CTCs capturing and enrichment, Figure 7(e). The advantages of very
fast processing speed (7.5 ml blood in less than 10 min) and the ability to collect more CTCs
from larger blood volumes make it very promising for a range of potential genomic and
transcriptomic applications.
Guan and co-workers 77 introduced a novel spiral micro-channel with a trapezoidal cross-
section and it showed a higher separation resolution than those with rectangular cross-section.
Their study found that particle focusing in spiral channel with trapezoidal cross-section is
sensitive to particle size and flow rate, and exhibits a sharp transition at a size-dependent
critical flow rate. Wu et al. 72 applied this novel spiral channel for the separation of
Leukocytes from blood sample. At the outlet, the larger white blood cells focus near the inner
walls, while smaller red blood cells are trapped at the core of Dean vortex near the outer wall.
An enhanced separation resolution and efficiency (>80%) was demonstrated. Later, isolating
CTCs from blood samples of cancer patients was demonstrated using the similar slanted
spiral micro-channels 71 by the same group, Figure 7(f). The device has successfully isolated
and recovered more than 80% of the tested cancer cell lines spiked in 7.5 ml blood within 8
min with high purity (400-680 WBCs ml-1; ~4log depletion of WBCs). In the initial clinical
investigation, the trapezoid chip successfully isolated CTCs from 10 out of 10 (100%)
patients with advanced stage metastatic breast and lung cancer (3–125 CTCs/ml), and it
allowed extensive heterogeneity studies via immunostaining and DNA FISH analysis.

22
Figure 7 (a) Schematic of a microfluidic flow cytometry, in which spiral channel is applied as a particle focuser
to provide high-throughput sheath-less 3-D focusing, followed by the downstream laser induced fluorescence
setup for particle detection and counting. Reproduced from Ref. 64 with kind permission from Springer Science
& Business Media. (b) Single cell encapsulation by droplets in a spiral channel. The spiral channel promotes
single cell stream ordering with a precise longitudinal spacing, and the downstream droplet generator
encapsulates cells into the droplets. Reproduced from Ref. 65 with permission from the Royal Society of
Chemistry. (c) Continuous triplet-particle separation in a spiral channel. Reproduced from Ref. 68 with
permission from the Royal Society of Chemistry. (d) Double spiral microchannel for tumour cell separation and
enrichment. Reproduced from Ref. 69 with permission from the Royal Society of Chemistry. (e) A multiplexed
microfluidic device stacked by three spiral channels for ultra-high throughput CTCs capturing and enrichment 76,

23
78
. (f) Ultra-fast, label-free enrichment of CTCs from blood samples by a spiral microfluidic device with
trapezoid cross-section. CTCs are focused near the inner wall due to the combination of inertial lift force and
Dean drag force, while white blood cells and platelets are trapped inside the core of the Dean vortex closer to
the outer wall 71.

3.3 Straight channel with pillar arrays or expansion-contraction arrays

Besides curvature, introduction of disturbance obstacles into straight channels will also
induce convective secondary flow, which was first reported to enhance mixing effects by
continuously splitting and redirecting fluid streams 24. By patterning a contraction-expansion
array on single side of a straight channel, Park’s group 79 successfully demonstrated three
dimensional single stream particle focusing under a sheath flow. At the entrance of the
contraction region, the centrifugal forces induce counter-rotating secondary flow, enveloping
a sample flow with a sheath flow in three dimensions. Besides particle focusing, a series of
particle separation in this kind of contraction-expansion array (CEA) channels were also
reported by the same group, including separating polystyrene beads of 4 µm and 10 µm in
diameter 53, blood plasma from red blood cells 80 and cancer cells from whole blood 81, Figure
8(a). Expansion-contraction array induces Dean-like secondary rotating flows, working
together with inertial lift forces on the suspended particles. Large particles or cells such as
cancer cells are influenced dominantly by the inertial lift forces, migrating towards
contraction-expansion side, while small particles or cells (e.g. red blood cells and white blood
cells) are dominated by Dean drag, shifting towards opposite side. Finally, it enables a size-
based particle/cell separation and enrichment 81.
The contraction-expansion array can also be patterned on two sides of the channel. Park et al.
82
investigated particle inertial focusing in a straight channel patterned with symmetrical
expansion-contraction arrays on both sides (multi-orifice microchannel), Figure 8(b). This
device is working without any sheath flow. The ordered particle distribution can be achieved
at central or side regions according to a particle Reynolds number Rp. Generally, particles are
focused at two side positions with Rp range of ~0.8 to 2.3 and at the centreline with the Rp
range of 3.0 to 3.5. The mechanism of these focusing patterns is relying on the combination
of inertial lift force and momentum-change-induced inertial force generated in the
contraction-expansion region. The trajectory mismatch between particles and fluid element
around the contraction-expansion region induces the lateral drift of the equilibrium position.
The extent of this lateral drift is variable according to particle size and flow rate. In the
Reynolds number range of 63 to 91, large polymer particles (~15 µm) were aligned at the
centreline of the outlet, whereas small particles (~7 µm) remained along both sidewalls.
Therefore, size-based separation of particles in this kind of multi-orifice channel is achieved,
and termed as Multiorifice Flow Fractionation (MOFF) 83. The MOFF has several advantages
such as continuous, label-free, sheath-less, and non-intrusive with minimal power
consumption. However, it is limited by the relatively low recovery yield. Although the
recovery yield may be increased by adjusting parameters such as the Reynolds number to
enhance central focusing, poor purity inevitably followed. Both high recovery yield and high
purity cannot be warranted at the same time. Therefore, Sim et al. 84 presented a multi-stage
multi-orifice flow fractionation (MS-MOFF), which is made by combining three multi-orifice

24
segments, 3 inlets, 3 filters and 5 outlets. It could improve recovery and minimize loss of
purity by collecting and re-separating non-selected particles from the previous separation.
The final recovery rate was successfully increased from 73.2% to 88.7% without significant
compromise on purity. By combining multi-orifice flow fractionation (MOFF) with
dielectrophoresis (DEP) techniques, Moon et al. 85 successfully separated human breast
cancer cells (MCF-7) from spiked blood sample, Figure 8(c). The inertial separation by
MOFF takes advantage of the high-throughput filtration of blood cells, and the serially
connected DEP separator works as a precise post-processor to further enhance the separation
efficiency and purity.
Diagnosis of malaria at the early stage of infection is challenging due to the difficulty in
detecting the low abundance of parasites from blood. Polymerase chain reaction (PCR)
method can be especially useful to detect low parasitemia levels, but the efficiency is
discounted by many factors, such as limited specificity of primers, presence of PCR
inhibitors in blood serum and DNA contamination from nucleated blood cells. Recently,
Warkiani and co-workers 86 presented an improved detection method for malaria by the
combination of inertial microfluidic processing and PCR detection, Figure 8(d). In the
symmetrical contraction-expansion channel, particles larger than 4 μm such as WBCs were
focused along the channel walls and isolated via the peripheral outlets. The malaria parasites,
on the other hand, were unfocused and distributed in all the three outlets. Therefore, enriched
and purified blood sample would promote more reliable and specific PCR-based detection.

25
Figure 8 (a) Inertial separation of particles by size in a CEA channel under the assistance of a sheath flow.
Reproduced from Ref. 53 with permission from Elsevier. (b) Continuous inertial separation in a multi-orifice
microchannel according to the size-dependent lateral migration termed as Multiorifice Flow Fractionation.
Reproduced from Ref. 82, 83 with permission from the Royal Society of Chemistry and American Chemical
Society Copyright (2009). (c) Continuous high-throughput separation of human breast cancer cells (MCF-7)
from blood cells by a combination of multi-orifice flow fractionation (MOFF) and dielectrophoresis (DEP).
Reproduced from Ref. 85 with permission from the Royal Society of Chemistry. (d) Improved detection of
malaria by the integration of inertial microfluidic processing and PCR detection 86.

26
In addition to the secondary counter-rotating flow within the cross-section of main channel
around the contraction-expansion region, a horizontal micro-vortex within the contraction-
expansion chamber can also be generated due to the detachment of boundary layer under a
high flow speed. Shelby et al. 87 was among the first to demonstrate a horizontal microvortex
in a diamond shaped cavity under high flow speed in microfluidics. This microvortex can
generate a rotational velocity as high as 12 m/s and a corresponding radial acceleration in the
order of 106 g. Such microvortex without any moving components are useful for the
investigation of effects of high radial acceleration on biological and chemical process, as well
as offering precise control and rotation of single cell 88, Figure 9(a). In their work, the single
cell was actively trapped and positioned at the centre of the microvortex by an optical
tweezer.
Later, Di Carlo’s group 89, 90 introduced the concept of “Centrifuge-on-a-chip” to selectively
isolate target particles/cells from heterogeneous background by the effects of inertial
migration and vortex trapping. The centrifuge chip consisted of a straight channel section and
symmetric expansion-contraction arrays, Figure 9(b). Rather than the optical tweezer, the
selective trapping and positioning of micro-particles are based on purely hydrodynamic
phenomena: the cross-stream inertial migration of micro-particles in the cavity expanding
area. The mechanism of trapping has been qualitatively explained by Mach et al. 90. Briefly,
the selective trapping process of particles is divided into three steps: (i) inertial focusing in a
straight channel, (ii) lateral migration around the expanding area, and (iii) circulation within
microvortices, Figure 9(b).
The functions of trapping, enrichment, labelling and solution exchange were demonstrated in
the Centrifuge chip 90-94. The most significant advantage of “Centrifuge-on-a-chip” is its
ability to selectively trap particles from the mainstream by size, which is believed as one of
the most size-sensitive separation methods 59. Di Carlo group has conducted a series of
investigation on its trapping sensitivity and efficiency through a variety of bio-samples,
including cancer cells spiked in blood 90, pleural fluids 92 and blood sample 93 from cancer
patients. One possible drawback of this device is that it basically works in a batch procedure,
and specifically effective in trapping of rare cells (e.g. CTCs), due to limited capacity in
expansion-contraction chambers. Wang et al. 91 proposed a modified microvortex-aided
device by adding side outlets in each chamber to continuously siphon larger particles from
chambers, Figure 9(c). However, the device is still limited by the capability for bimodal
separation with a single size cut-off and well-defined size difference. And it will become
challenging when treating real-world samples that often includes heterogeneous mixtures of
multiple particle components. Wang and Papautsky 95 developed a multi-modal inertial sorter
that consisted of a long straight section with series-connected separation units. The authors
successfully demonstrated triple-sized particle separation with high resolution by sequencing
two separation units and adjusting proper system parameters.

27
Figure 9 (a) Rotational control of single cells by microvortices in a diamond-shaped chamber. Reproduced from
Ref. 88 with kind permission from Springer Science & Business Media. (b) Size-selective trapping of circulating
tumor cells using “Centrifuge-on-a-chip”. Reproduced from Ref. 93 with permission from the Royal Society of
Chemistry. (c) Vortex-aided inertial microfluidic device for continuous sorting of particles by size. A modified
side outlet in each chamber siphons the trapped large particles out continuously, meanwhile smaller particles
exist from the main channel. Reproduced from Ref. 91 with permission from AIP Publishing LLC. (d)
28
Engineering flow deformation by programing a sequence of cylindrical pillars within a straight microchannel in
inertial microfluidics and its application on synthesis of shaped polymeric microfibers. Reproduced from Ref. 26,
96
with permission from Nature Publishing Group and John Wiley and Sons .

Besides expansion-contraction chamber, a micro-pillar within a micro-channel will also

induce irreversible twisted flows at a finite inertial flow. Furthermore, the lateral position of
the pillar can be used to tune the position and shape of the net recirculating flows across the
channel. It could enable precise control of fluid transformation by programming the positions
of sequenced cylindrical pillars, Figure 9(d). To explore the capabilities of this sequenced
pillar microchannel, Amini et al. 26 have successfully demonstrated the sculpture of cross-
sectional shape of a laminated stream into complex geometries, moving and splitting a fluid
stream, solution exchange and particle separation. Nunes et al. 96 utilized this microfluidic
technique to fabricate polymeric microfibers with noncircular cross-sectional shape, Figure
9(d). The computer-aided design (CAD) tool uFlow has a stored library of pre-computed
fluid deformations that are produced by individual pillars in the flow channel. It is a useful
tool to design the pillar sequences, and to predict the shape of net flow deformation. The
cross-sectional shapes of various fabricated microfibers agreed reasonably well with that of
predicted using the uFlow code.
Similar to the secondary flow induced by the contraction-expansion chamber or channel
curvature, the locally induced secondary flows by sequenced pillars could also be used to
modify the inertial migration progress. Chung et al. 97 demonstrated single-stream focusing of
microparticles through controllable cross-stream migration, which was aided by the locally
tuned secondary flows from the sequential micropillars. Besides, programmable flow
deformation in sequential micropillars was used to conduct solution transfer around particles
or cells. The microfluidic device was applied for a functionalized bead bioassay, achieving
high-yield and continuous separation of biotin-coated beads from extra FITC-biotin, as well
as extraction of leukocytes from lysed blood 98.

3.4 Serpentine channel

As we know, in spiral channels, the curvature is along a single direction, therefore secondary
flow can reach steady state after a given channel length, and almost consistent within
different cross-sections. Analysis of particle behaviour can be approximated by the static
superposition of inertial lift force field with a secondary flow field. However, in a serpentine
channel with alternating curvatures, the situation becomes more sophisticated. For example,
with alternating curvatures, secondary flow may not approach steady state after each turn, the
same with the movement of particles, and accumulation of this unsteady may cause
unpredictable and non-intuitive behaviour of particles. Therefore, analysis of particle
behaviour by the superposition method may not give a convincing explanation.
Di Carlo et al. 43 investigated the effects of alternating curvatures on the particle inertial
migration. In their study, original four equilibrium positions in a straight channel with square
cross-section were reduced to two in a symmetric serpentine channel due to the symmetry of
the system. Above a critical Dean number, focusing was perturbed. Furthermore, in an
asymmetric serpentine channel, the number of equilibrium positions could be further reduced
to one, focusing again became more complex as Dean number increased, Figure 10(a). From
29
their understanding, the balance between inertial lift forces (FL) and Dean drag force (FD)
determines the preferred location of focusing positions. Dean flow does not create particle
focusing, but it acts in superposition with inertial lift forces to reduce the number of
equilibrium positions created by the inertial lift forces. If FD>> FL, then no focusing will be
observed, and if FD<<FL, then focusing due to inertial lift forces alone will be observed. The
2𝑅
ratio of inertial lift force to Dean drag was scaled as: 𝐹𝐿 ⁄𝐹𝐷 ~ (𝑎⁄𝐻 )3 𝑅𝑒𝑛 , (𝑛 < 0). This
𝐻
relation suggests a strong third-power dependence on the ratio of particle to channel
dimensions. Even at the same Reynolds number, small particles may be still unfocused
independent of channel length, because of dominant FD, while larger particles quickly
become focused. Employment of this mechanism, an inertial filtration device using an
asymmetrical serpentine channel was developed and tested 99. Within the expectation, large
particles were well focused and small particles below a threshold remained unfocused and
randomly distributed. Therefore, large particles were completely removed from the mixture,
leaving behind small particles with a high purity (90%-100%). However, because small
particles are still unfocused, plenty of them will enter the reservoirs meant for large particles,
leading to unsatisfactory purity of large particles’ collection.
Inertial focusing in an asymmetric serpentine channel can produce well-defined particle
focusing as well as highly regulated inter-particle spacing, which is very promising in a range
of applications, e.g., the flow cytometry system 100, particle active sorting 101, hydrodynamic
stretching of single-cells 102 and bioparticle concentration 103. Particle inertial focusing may
replace the hydrodynamic focusing unit in the standard commercial flow cytometry,
composing a compact microfluidic flow cytometry system 100. Besides, it could also be
integrated with ultrafast optical capturing system for automated flow-through single-cell
image analysis 104. For particle sorting, Toner’s group 101, 105 developed a hybrid microfluidic
device which combined deterministic lateral displacement (DLD), inertial focusing and
magnetophoresis (MP) to isolate rare circulating tumour cells (CTCs) in an antigen–
dependent and independent way, Figure 10(b). In this device, after the sample of raw blood is
introduced into the chip, the blood cells first encounter an array of circular pillars with a
specific gap, where DLD size-based hydrodynamic force filtrate smaller red blood cells
(RBCs) from white blood cells (WBCs) and CTCs with much larger size. Subsequently, the
mixture of WBCs and CTCs flows into an asymmetric serpentine microchannel where they
are rapidly aligned along a single path by the inertial effects. At the end of the serpentine
channel, a strong magnetic field is applied to deflect those cells that are magnetically labelled.
The CTCs can then be collected separately.
Besides asymmetric serpentine channel, our group have investigated the focusing patterns
and corresponding mechanisms in symmetric serpentine channels 106-108. Three focusing
patterns were observed by increasing the flow rate: (i) two-sided focusing, (ii) the transitional
focusing, and (iii) single central focusing. From our understanding, the secondary flow due to
the channel curvature, not only has mixing effects, but also can focus particles at the channel
centre if the vertical component of secondary flow which is responsible for the mixing effects
is suppressed by reducing the aspect ratio of channel 106. After that, particles will be focused
at the channel lateral centre if the secondary flow is greater than the inertial lift force. In

30
contrast, if inertial lift force is stronger than the secondary flow, particles will be focused near
two sidewalls. If they are in the similar order, particles will be within a transitional focusing
pattern, where particles are focused as a wide streak. Employing the overlap of two-sided
focusing pattern for small particles and single central focusing pattern for large particles
would enable a complete separation by differential lateral equilibrium positions for binary
particles 108, Figure 10(c).
A DEP-inertial coupled microfluidic device was proposed through the combination of inertial
lift force and DEP force 109. This combination can modify the inertial focusing patterns in a
serpentine channel with a vertical DEP force, which was implemented by interdigitated
electrodes patterned on the bottom of microchannel. By levitating particles towards the
vertical centre of channel by DEP force, the original two-sided focused particles will migrate
towards the lateral centre, which indicates that increasing lateral component of secondary
flow drag as well as reducing its vertical component will promote a single central focusing of
the particles. This phenomenon further proves that our explanation about inertial focusing in
serpentine channels is reasonable. Moreover, this device demonstrated the possibility of
combining passive inertial focusing with active DEP to enable more controllability as well as
maintaining the advantages of high-throughput.

31
Figure 10 (a) Inertial focusing of particles in serpentine microchannels. In a straight channel with square cross-
section, focusing of particles into four single streamlines is observed, with each of them facing the centre of
each wall. For a symmetric serpentine channel, the symmetry of system reduces focusing to two streams. In an
asymmetric serpentine channel, focusing down to a single stream is observed. Reproduced from Ref. 43.
Copyright (2007) National Academy of Sciences, U.S.A. (b) A hybrid microfluidic device which integrates
deterministic lateral displacement (DLD), inertial focusing in asymmetrical serpentine channel and
magnetophoresis (MP) to isolate rare circulating tumour cells (CTCs) in an antigen–dependent and independent
way. Reproduced from Ref. 101 with permission from the American Association for the Advancement of Science.
(c) Size-based separation of binary particles by their differential equilibrium positions in a symmetric serpentine
microchannel. Reproduced from Ref. 108 with permission from Nature Publishing Group.

4 CONCLUSIONS AND OUTLOOK

In this review, we first discussed the fundamental dynamics of particle movement. Several
lateral forces might act on particles in an inertial microfluidic device. Viscous drag force,
arising from the velocity difference between fluid element and particles, and basically retains
32
the particles within fluid streamlines. Lateral forces, such as Magnus force, Saffman force,
shear gradient lift force and wall lift force are perpendicular to the main streamlines, and
account for the intriguing phenomenon of particle inertial migration. In a wall-bounded
Poiseuille flow, Magnus and Saffman force are normally neglected. And the balance of
centre-directed wall lift force and wall-directed shear gradient lift force can well explain the
experimental observation by Segre and Silberberg 12, 13, 17. Besides, the properties of particles
(e.g. deformability, shape, size) and the fluid property (e.g. viscoelasticity) may also
influence the lateral migration of particles 45, 49, 110.
Subsequently, a comprehensive review of recent progress of the inertial microfluidic
technology was presented, according to the structure of functional microchannels, i.e. straight
42, 44, 49, 54
, straight channel with pillar arrays 26, 97, 111 or expansion-contraction arrays 25, 79, 82,
spiral 77, 112, 113, and serpentine 43, 106, 114. These inertial microfluidic devices have been widely
explored for their applications in biomedicine and industry, such as extraction of blood
plasma 80, 107, separation of particles and cells 53, 68, 72, 108, 115, solution exchange 98, 116, 117, cell
enrichment 118, isolation of circulating tumour cells (CTCs) 69, 71, 75, 81, 90, 92, 93, 101, detection of
malaria pathogen 86, 119, microfiber fabrication 96, cell cycle synchronization 73, cell
encapsulation 65, and hydrodynamic stretching of single cell 102, 120 etc. There are also several
commercial/quasi-commercial prototypes reported based on the inertial microfluidic
techniques 35, e.g. CTC-iChip by Johnson & Johnson 101, ClearCell FX system by Clearbridge
BioMedics 75, 121, mechanophenotyping platform by CytoVale 122, as well as Microfluidic
“centrifuge-on-a-chip” by Vortex Biosciences 90, 123.
Although there have been extensive investigations on the mechanism of particle inertial
focusing, and a variety of applications on particle/cell separation have been demonstrated,
quantitative design rules are still lacking for particle inertial manipulation in different shaped
channels. Therefore, more dedicated work is needed to uncover the detailed underlying
mechanism, and provide explicit rules to the end users even without specialized knowledge
and experience about inertial microfluidics. In addition, more efforts are needed to improve
the separation resolution and processing speed through optimization of channel structure and
scale up scheme. Although some parallelization designs can provide good examples 58, 76, 115,
124
, a general design guidelines and optimization scheme based on different channel structure
would be more helpful.
Combination of passive inertial microfluidics with active manipulation techniques will enable
more versatile and powerful functionality. Several manipulation techniques may be
connected in series with independent physics, or in parallel with coupled physics. Ozkumur et
al. 101, 105 reported an integrated microfluidic device which combined deterministic lateral
displacement (DLD), inertial focusing and magnetophoresis (MP) to isolate rare circulating
tumour cells (CTCs) in an antigen–dependent and independent way. Meanwhile, Moon et al.
85
successfully separated human breast cancer cells (MCF-7) from spiked blood sample by a
combination of multi-orifice flow fractionation (MOFF) and dielectrophoresis (DEP)
techniques. In both works, although the physics in each section is common and independent,
actually they are weakly linked, because the interface between each section, such as flow rate
or pressure, needs to be balanced carefully, so that each section can work normally.

33
We proposed a DEP-inertial microfluidic device, where negative DEP force and inertial lift
force are coupled along the vertical direction to adjust the focusing positions of the particles
in three dimensions 109. Based on this platform, the DEP force could be used as a force probe
to measure the magnitude of inertial lift force. Besides the DEP force, magnetic, acoustic or
other active forces may also be combined with inertial lift force in inertial microfluidics, to
achieve more versatility and flexibility for Microfluidic system.
Current study on particle manipulation by inertial microfluidics is often limited for particles
within micro-scale region. The territory of nano-scale particles (DNA, protein and virus etc.)
has not been explored yet in inertial microfluidics. For nano-scale particles, the Brownian
motion becomes more obvious, and maybe even overcomes its inertial effects. Therefore, a
reduced channel size (sub-micro to nano-scale) may be not a necessary consequence, and
introduction of additional viscoelastic effects could be a possible alternative 125.
To date, inertial focusing has been conducted where particles are suspended within liquid
phrase. There is no reported work for particle-air system, i.e. aerosol. No matter from the
aspects of fundamental exploration on particle movement within micro-airway, or from the
aspects of its potential application in biomedicine and industry, e.g. understanding
functionality of respiratory system, inhalation drug delivery and air pollution purification etc.,
this kind of research is still very meaningful.
In conclusion, although significant progress has been achieved for inertial microfluidics over
the last decades, even with a few commercial prototypes developed, inertial microfluidics
itself is still in its early development stage. A wide region of its raw territory has been waiting
for exploration and exploitation. We believe that in the near future inertial microfluidics
remains a hot research topic with expanding applications, because of the superior advantages
of the technology, such as high throughput, simple structure and low cost.

ACKNOWLEDGEMENTS

This work was partially supported by the University of Wollongong through a UIC grant and
China Scholarship Council.

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