Microfluidics:

Anika Agarwal, Vaishnavi Kishore, Jeremy Mathews, Blake Steines, Zach Trotter

The study of Microfluidics involves the analysis of fluid behavior on a sub-millimeter

scale. When moving from the macro to the micro scale, unique flow conditions begin to manifest
- namely the domination of surface tension, energy dissipation, and fluidic resistance. A key
change in fluidic systems at the microscale is the move towards a lower Reynolds Number. The
Reynolds Number is the ratio of inertial to viscous flow in a fluid. As this quantity begins to
lower, co-flowing fluids begin to exhibit laminar rather than turbulent flow, and molecular
transport between these layers begins to occur by diffusion. The aim of studying Microfluidics is
to better understand how one can manipulate molecules through space and time.
To date the most commercially successful use of Microfluidics has been its use in inkjet
printers. Over time, the technology has also been applied to spur advancements in the molecular
biology industry. Most notably, microfluidics is simplifying the process of enzymatic and DNA
analysis by integrating the multi-step processes onto a single chip. This has not only made the
process of analysis much easier, but has also allowed for the process to occur with less material
costs and in a greatly reduced amount of time. Microfluidics is also being used on the medical
front in lab-on-a-chip technology, which is leading to advancements in personalized medicine.
Microfluidic systems rely on two primary types of fluid transport. The first, pressure-
driven flow, is similar to the way pressure drops affect fluids at larger scales. The fluids are
forced towards the area of lower pressure. In microfluidics, this type of flow can even be
mathematically explained with the same equation that larger scale models use. This equation
would be the Navier-Stokes equation. The main problem with this type of flow within the
context of microfluidics is that the manufactured pressure often needs to be much higher than the
atmospheric pressure. For this reason, pressure-driven microfluidic flow is generally impractical.
For this reason, electrokinetically-driven flow is much more common when experimenting with
microfluidics.
There are two main types of electrokinetically-driven flow: electroosmosis and
electrophoresis. Both types of flow can be used at the same time, such that the flow velocities are
summed to find the total electrokinetic mobility of each molecule. Electroosmosis is the
movement of an aqueous solution due to an applied electric field. There also needs to be a
charged double-layer where the liquid interacts with the solid channel, which is created due to
the attraction between bound surface charges and ions in the solution, which flow past them.
When the flow has a low Reynolds number, meaning theres steady, incompressible flow, the
Navier-Stokes equation can be used to derive the Helmholtz-Smoluchowski equation (U=),
which characterizes the velocity of the electroosmotic flow based on permittivity (epsilon),
electroosmotic potential (EO zeta), the axial electric field (E x), and the viscosity of the fluid
(mu). While electroosmotic flow is unidirectional, meaning the fluid only moves in one direction
towards a single electrode, electrophoretic flow is bidirectional. This is because the solution is
actually stationary, and its actually the charged particles within the solution that move towards
either electrode. The direction in which they move is based on the charge that the particle carries,
with positive ions flowing towards a negative potential and vice versa. Because of this
phenomenon, a net neutral solution would not result in bulk motion since an equal amount of
particles would flow in opposite directions. The velocity of flow for electrophoresis can also be
described by the Helmholtz-Smoulchowski equation, albeit with some minute changes. Since the
flow is bidirectional, the equation becomes positive compared to the equation for electroosmosis.
There is also a different potential term, (EP zeta potential), which is a property of the charged
dye particles found in the solution as opposed to (EO zeta potential), which is a property of athe
capillary surface. These terms will not
normally be equal. The resulting equation
becomes (U=) for the velocity of
electrophoretic flow. Both of these equations
do not relate velocity to a radial position,
which is why both types of electrokinetic
transport result in what is known as plug-like
flow, a profile of which can be seen in figure 1.
These images prove that the velocity is
constant across the channel, which agrees with
the Helmholtz-Smoluchowski equation.
In microfluidics and its many applications, we seek to understand how mass is transferred
on micro and nano-level scales. Many of the same principles that govern mass transport on
macro scales still apply on micro scales, but can result in interesting properties that would not be
seen on larger scales. These interesting properties are due to the characteristics that arise from the
high surface area to volume ratios that exist in microfluidic devices. In this discussion of mass
transport, it is assumed that flow is only driven by a competition of pressure differences and
viscous force and that flow is incompressible, which makes for an easy analogy with an electric
circuit. There is a wide array of particles for which their mass transport is studied in microfluidic
and these include: ion and small molecules (1 1nm), micelles and small proteins (1 10nm),
colloids and viruses (10nm 1um), and cells and bacteria (1um 10um).
 The movement of these nano and micro-particles can be broken into two main categories:
convection and diffusion. Figure 1 is a good pictographic representation of the nature of
convection and diffusion. Convection is simply the movement of
a particle in a straight line. Another way of saying this, is that
the particles movement is direction. The driving force for a
particle to move in a given direction like this is usually due to a
pressure difference. In microfluidics, this pressure difference is
usually created by a pressure pump, which is applied to the
closed channels of the microfluidic chip. The length
that a particle travels by convection is proportional to
the time over which the particles distance is being
observed, which is given by equation (1). Diffusion,
on the other hand is the non-directional, random
movement of these particles. It is caused by thermal
agitation amongst the particles which can be
modeled by a random walk, in which, at any given
time, a particle has each chance of moving up, down,
left, right, forward and backward. The length that a
particle travels by diffusion is proportional to the
square root of the time over which the particles
distance is being observed, and is given by equation (2). The proportionality constant for
movement of particle by diffusion is called the diffusion coefficient of a particle (D). The
diffusion coefficient is larger when the particles are at a higher temperature, and lower when the
particles are of a higher viscosity, which is made clear in equation (3), also known as the Stokes-
Einstein Law. The numerator of equation (3) can be thought of as the motor for diffusion, and
the denominator can be thought of as the friction for diffusion. In order to better understand
many microfluidic systems, engineers utilize a dimensionless constant that relates the diffusion
time to the convective time. This is a ratio called the Peclet number, which is given by equation
(4). In a model that only considers diffusive and convective movement of particles, we arrive at
the conservation equation, which relates the concentration of the particle in the fluid to the
movements of the particles (equation (5)).
 One common construction in many microfluidic applications is called coflow. It is
essentially two, side by side, laminar flows of two different solutions that mix by diffusion
(Figure 2). Using equation (2), we can figure
that the time for one of the solutions to diffuse
into the other is the channel width (w) squared,
divided by the diffusion constant of that
solution. Using this time in the convection
equation, we can derive that the length of
channel needed to fully diffuse a solution in
coflow is the channel width times the Peclet number. In coflow, assuming a model that only
considers convection and diffusion, we can equate the convective contributions to the
concentration change in the x direction with the diffusive contribution to the concentration in the
y direction (equation (6)). Equation (6) can be solved for different initial conditions to study
different things in a coflow setting such as: kinetics of chemical reactions, kinetics of protein
folding, etc.
In the coflow configuration described previously, diffusion of two fluids is a rather slow
process. In an effort to overcome the long time needed for two fluids to mix together in a
microfluidic system, engineers have developed many methods to expedite the process of mixing.
In a normal coflow configuration, the length of channel needed for efficient mixing by diffusion
is the Peclet number times the channel width (Table 2, 1). This length is drastically shortened by
modifying the microfluidic chip. One way to do this is by applying pressure on both sides of a
stream of fluid to reduce the diameter of the stream, thus making the effective channel width
much less. Another effective way to mix fluids is by multilaminating the flow, so instead of
having two fluids flowing in a laminar fashion side-by-side, you can run the fluid through a
complex matrix which effectively makes a comb out of the fluid with many streams side-by side
alternating fluids with each tooth of the comb. This multilamination technique reduces the
orginal length required for mixing by a factor of N2 where N is the number of tooths in the comb
of fluids (Table 2, 2). Another popular technique for mixing fluids is called grooving. This is
where divots are made in the wall of the microchannel which causes the once laminar flow to
become chaotic and semi-turbulent. The new length required for mixing in grooving flow is
diminished to the channel width times the natural log of the Peclet number (Table 2, 3). One final
method that is often employed to mix fluids is by using oil droplets. The method involves inject
multiple fluids inside an oil droplet which drastically confines the space of the fluids which
greatly enhances the mixing speed. The length of channel needed to mix fluids through this
method is the same as the length needed for grooving flow (Table 2, 4).
The study of microfluidics and its application to the biomedical engineering field has
allowed for many advancements in medicine through a more thorough understanding of the
functions of the body. The use of microfluidics in microchips is accomplished through the use of
microanalysis, biodefense, and microelectronics. Microfluidics flow is laminar due to the tiny
volume being moved, and yet the viscosity of the fluid remains constant allowing for the use of
the Navier stokes equation. These key features allows for in-vitro testing of microfluidics to be
used as a study for in-vivo application of many medical phenomena.
In order to understand the applications of microchips, the knowledge of the different
materials used to create them must be obtained.
This use of different materials allows for the
testing of different types of cells and different
locations of the body, since there are many
differences across the body. One of the first
materials used to design microchips was silicon
and glass. When microchips were first created,
silicon was greatly used in microelectronics and
the silicon industry was thriving so it only made
sense to design these microchips using the
inexpensive material. Silicon also carried very unique attributes that contributed to its use. The
high thermal conductivity allows for silicon to to pass high currents through itself without
harming the material itself. This allowed the first microchips to be very effective and allow for
preliminary learning of basic bodily functions. Silicon also had a high stability and compatibility
to solvent, allowing for the microchip to be submerged in fluids such as interstitial fluid,
increasing its application of testing to more specific areas of the body while maintaining its
selected properties. One issue that arose from using silicon was that at the micro level, scientists
wanted to observe the exact chemical reactions taking place and the molecular movement of
ions. Silicon did not allow for visibility and so engineers decided to use glass because of its high
transparency, allowing them to see what was actually happening as the microfluidics passed
through the chip. Glass was a good replacement because on top of the transparency, it maintained
the solvent compatibility and high thermal conductivity of silicon. Glass also has a high
resistance to pressure allowing for its use to be applied in many different scenarios. The only
drawback to using glass for microchips was that it was more expensive that buying cheap silicon.
Glass was also chemically inert which was a positive aspect as well drawback due to the fact that
this limited its application to some parts of the body.
Polymers have become on of the front runners in the microchip choice of material. Their
low cost combined with the high levels of compatibility and bio-chemical applications make
them some of the most perfect materials to use. Another plus side to polymers is that different
polymers can be used to achieve different end products for measurement. One of the most
commonly used polymers is Polydimethylsiloxane (PDMS). Due to its high use and testing, it is
also used in many products that come in contact with the body since it has a proven
biocompatibility. Like glass, PDMS also have a high transparency allowing for the microfluidic
flow to be seen with the naked eye. This polymer also has a high elasticity and permeability
which are two of its more important aspects. The elasticity allows for PDMS to be used in
various applications and to be able to adapt to changes in the environment with ease while
maintaining its chemical properties. PDMS is specifically gas permeable which allows it to be
applied to cell study. PDMS is used to test the lining of alveoli as well as parts of the stomach in
order to test the diffusion of gas molecules through the membranes. This application has also
been used in gas sensors since PDMS can be used to test minute, micro sized changes in gas
concentrations as well as the appearance of new gas particles that will have different
permeabilities through this polymer. PDMS does however attract and absorb hydrophobic
molecules which can cause discrepancies and issues in testing. This fact paired with its lack of
compatibility with organic solvents causes PDMS to be restricted to testing in aqueous
environments. The submersion in aqueous environments also allows for the polymer to be
preserved as long as possible since it loses its chemical properties over long periods of time.
There are other polymers used in engineering design of these microchips, they are just in less
quantity. Many laboratories use Polystyrene (PS) in cell culture tests due to the ideal surface
which is has. Being both biocompatible while maintaining rigidity and inertness allows for this
polymer to be tested while maintaining its structure. This comes in handy when testing new drug
compounds and other medical implants. Another unique property of PS is that it can be
chemically altered to have a hydrophilic surface if needed. This allows for increased testing in
laboratories before moving to animal testing. Polycarbonate (PC) is a type of polymer that has
one of the highest thermal resistivities. This has come in handy when testing substances through
microfluidics at largely varying temperatures. PC can withstand major fluctuations as well as
extreme temperatures while maintaining its chemical properties.
Alongside polymers, paper is also an extremely cheap material used in the design of
microchips. Paper is also environmentally friendly which allows for simplification of disposal. The
design of a microchip on paper allows for easily manipulated designs that are lightweight and can be mass
produced. Paper microchips also have a high biocompatibility, not like that of polymers, but still
applicable to external body testing. Paper microchips has been used in many different types of
microfluidic testing such as pregnancy testing, HIV diagnosis, glucose blood sensor, and drug testing. The
easy access and disposability, alongside the eased transportation and flexibility allows for paper based
microchips to be used in developing countries as well. Many organizations like doctors without borders
use these cheap microfluidic devices to test large numbers of people for many things at once. The only
drawback to using paper of any of the other materials is that fact that it is not perfectly precise and
accurate in its design. If one thousand paper chips were created, they would not all be identical due to the
loss of this precision in using paper. But the balance of the cheapness and accessibility makes up for this
in preliminary testing. Past the point of preliminary testing, paper becomes less and less usable because
the readings and output will not be as reliable, plus the doctors and engineers can afford to use better,
more expensive material once this point has been reached.
Hydrogels are an important use of polymers in a material for microchips. Sodium Polyacrylate is
used in the polymeric chains with water to create many of the hydrogels used in microfluidic testing. Its
malleability along with its non-toxicity allows for it to be molded and shaped into many parts of the body
while maintaining its properties. The most unique and applicable factor of hydrogels is the creation of a
diffusion matrix similar to that of the bodys cells. This allows for different drugs, substances, and
nutrients to be observed as they would be in the body. With the hydrogel, the diffusion of drugs into and
out of the blood stream becomes much more easier. By passing micro volumes of fluid through these
chips and detecting the various changes in concentration as well as seeing the diffusion occur allows for
doctors to better understand and visualize how the body works on the cellular level.
The application of microfluidics in microchips has allowed for great steps forward in medicine
and engineering. Some applications are more simple such as pH monitoring and the administration of
drugs. Other applications are much more in depth and complicated. In order to measure the pH of fluids
passing through the microchips, solid state sensors are placed inside the circuitry of the microchip which
allows for high precision readings at each point in testing. This is very important due to the fact that pH in
the body must be maintained in different areas as to keep the body functioning. When testing new drugs,
the doctors and engineers must know how the drug affects the pH of the body and how to maintain
physiological pH. Microfluidics also allows professionals to see the effect of drugs on the cells of the
body. By viewing the effects on a molecular level, doctors can then tell what exactly is happening with
each drug. This process has allowed for great advancements in medicine in the last 15 years. With this
advancement, comes the need for more efficient drug administration, which can also be provided by
microfluidics. The use of laboratories-on-a-chip has allowed for a more efficient drug synthesis and
administration. This cuts down the need for oral ingestion or topical application, while reducing the risk
of infection and pathogens that comes from a more invasive type of drug administration. Also, the
knowledge of how the drug acts in the body, has allowed for a specialized drug path to be created to allow
for faster application, which leads to faster reaction with the body, which leads to faster help for the
patients. One of the biggest and more complex advancements using microfluidics is the study of the C.
elegans organism. C. elegans are a very simple worm that is transparent allowing for easy study. The
reason doctors were very interested in this organism, is that while the organism is extremely simple,
allowing for easy study, it still has a fully functional nervous system. The study of this organism has led
to large advancements in our understanding of the neural function of humans and other animals. The
largest issue with studying these organisms was the ability to take high resolution images. To do this, the
movement of the C. elegans had to be restricted, to slow them down and allow the cameras to focus.
Other techniques required freezing, glue or other harmful substances that induced slowed movements.
The use of microfluidics has solved this long time issue without harming the organism itself. By
streamlining these C. elegan into microtunnels, the area for movement allowed for these worms is greatly
decreased. By decreasing the area they have to move, their movement is slowed down enough for the
camera to increase its resolution by great magnitudes. Another area that microfluidics is becoming more
and more useful is cell analysis in cell interactions with its environment. Using the microfluidic laws of
mass and fluid transport, doctors are able to study cell cytometry and determine toxins and stimuli for
different cells. This is accomplished due to the ability of great control of flow in microfluidics.
Microfluidics allows for controlled research that can help in understanding the physics of flow
inside and outside the human body. Some of the largest advancements in microfluidics today are in
medical research, because microfluidic chips are able to mimic many body conditions that are otherwise
difficult to re-create. These microchips and microfluidic systems have historically been used in patient
point-of-care testing. In the more current research, these chips will be used to create more personalized
medicine through organ-on-a-chip devices. These microchips can be created for each patient so that their
own cells may be used to test treatment methods for diseases. Microfluidics is a vastly applicable field
that is still growing.
Questions:
1. Which of the following is not correct about electrophoretic flow?
a. Charged particles flow in either direction based on the charge they
hold.
b. One could expect a velocity profile to display plug-like flow.
c. The solution must be able to flow in either direction to achieve this
type of flow.
d. The velocity equation is a variation of the Helmholtz-
Smoluchowski formula.
e. Electrophoretic flow does not have to be independent of
electroosmotic flow.
2. The time it takes for 1 solution to diffuse into another through coflow is
proportional to:
a. (width of the channel)2/(diffusion constant)
b. (width of the channel)/(diffusion constant)
c. (width of the channel)/(diffusion constant)2
d. (width of the channel)2/(diffusion constant)
e. (width of the channel)2/(diffusion constant)2
3. One of the leading applications of microfluidics is the use of microchips to study
the effects on cells in an environment similar to an in vivo setting. Which of the following
is not a material used to create these microchips?
a. Carbon-fiber
b. Hydrogels
c. Paper
d. Silicon
e. Glass
f. Polymers

References
 1. Molho, J. I. (n.d.). Fluid Transport Mechanisms in Microfluidic Devices.
Retrieved May
9, 2017, from http://mems.stanford.edu/~aeh/publications/Molho_asme98.pdf
2. http://www.elveflow.com/microfluidic-tutorials/cell-biology-imaging-reviews-
and-tutorials/microfluidic-for-cell-biology/microfluidics-applications-a-short-review/
3. http://www.lof.cnrs.fr/IMG/pdf/Salmon_Agay.pdf