BIOMEDICAL
Annals of Biomedical Engineering ( 2020)
https://doi.org/10.1007/s10439-020-02526-9
Original Article
Plasma Isolation in a Syringe by Conformal Integration of Inertial
Microfluidics
JUNG Y. HAN and DON L. DEVOE
Department of Mechanical Engineering, University of Maryland, College Park, MD 20742, USA
(Received 27 December 2019; accepted 28 April 2020)
Associate Editor Tingrui Pan oversaw the review of this article.
Abstract—A thermoplastic microfluidic substrate is confor-
mally integrated onto the cylindrical barrel of a conventional
venipuncture syringe, forming a spiral inertial separation
element supporting the isolation of plasma from diluted
whole blood. The cylindrical shape of the syringe itself serves
to define the flow path required for inertial separation by
transforming a linear microchannel to a spiral topology. The
hybrid system enables inertial plasma separation by Dean
flow focusing within the same syringe used for a patient
blood draw, with the seamlessly interconnected microfluidic
element operated by automated or manual actuation of the
syringe plunger. Plasma isolation is achieved without the
need for external instrumentation. Device design and fabri-
cation challenges are discussed, and effective plasma isola-
tion within the system is demonstrated, with a peak
separation efficiency above 97% using 25 9 diluted blood.
Keywords—Dean flow focusing, Inertial separation, Solvent
casting.
INTRODUCTION
Lab-on-a-chip technology is an important enabler
for a broad range of diagnostic and clinical platforms.
The miniaturization and integration of analytical
functions into a single microfluidic chip represents a
powerful concept offering numerous advantages for
increasing performance, reducing cost and complexity,
and enhancing accessibility of biomedical assays.
Microfluidic devices typically consist of discrete com-
ponents that must be interfaced with external instru-
mentation and fluidic interconnects for sample
introduction from upstream assay stages, sample
delivery to downstream stages, reagent addition, and
Address correspondence to Don L. DeVoe, Department of
Mechanical Engineering, University of Maryland, College Park,
MD 20742, USA. Electronic mail: ddev@umd.edu
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power delivery. This requirement presents a world-to-
chip interface challenge that is especially acute for
systems designed to operate in resource-limited clinical
environments, where maximizing device reliability,
minimizing operational complexity, maintaining device
sterility, and reducing the potential for exposure to
biological hazards are critical concerns.5,27,30,31
An alternative approach to interfacing microfluidic
chips with supporting components for fluid handling
and power delivery is to directly integrate microfluidic
functionality into existing clinical tools. In this way,
microfluidics can add new capabilities such as sample
preparation or analytical steps to established instru-
mentation without imposing the need for additional
interconnections or specialized infrastructure for de-
vice operation. Here we explore the integration of a
microfluidic sample preparation element into a dis-
posable plastic syringe as one of the most ubiquitous
biomedical devices. Blood sampling by syringe
venipuncture is routinely performed as the first step for
blood-based diagnostics and serological biomarker
screening.43 For molecular diagnostics such as
immunoassays, sampled whole blood is typically sep-
arated by centrifugation to isolate plasma as an ana-
lytic matrix, thereby eliminating interference imposed
by blood cells on target analyte detection.32 While
centrifugation is a straightforward process, it requires
access to laboratory equipment that may not be readily
available in smaller clinics, field environments, and
other resource-limited settings. A variety of small-scale
technologies have been developed to perform cen-
trifugation with manual power input,2,3,44,51 but like
conventional contrifuges these platforms add multiple
process steps and components, thereby increasing as-
say complexity. To overcome the limitations
of centrifugation, a range of microfluidic techniques
 2020 Biomedical Engineering Society
 J. Y. HAN
based on active8,14,15,19,21,23,26 and pas-
7,12,13,22,28,33,34,37,39,40,45,52
sive separations have been
reported for plasma isolation in planar chips, including
the development of power-free microfluidic devices
capable of isolating plasma from whole blood using a
combination of vacuum pumping and sedimentation-
based cell separation.49 While these microfluidic tech-
nologies offers advantages over centrifugation for
effective plasma isolation from small volume blood
samples, they still require the use of separate compo-
nents for sample collection and plasma separation.
Here we investigate the integration of microfluidic
plasma separation elements into standard thermo-
plastic syringes, resulting in a seamlessly-integrated
device combining initial blood sample collection and
plasma processing. Passive separations that employ
hydrodynamically-driven flow fields are particularly
appropriate for syringe integration, since flow control
may be achieved through manual or automated actu-
ation of the syringe plunger. Plasma isolation is
achieved using Dean flow focusing (DFF), an inertial
focusing method that offers high throughput passive
particle separation resulting from the summation of
inertial lift force and Dean drag force during particle
transport in a curved microchannel.6,9,11,18,24,28,33,42,45
While conventional microfluidic DFF devices are
routinely fabricated in planar
chips,1,6,9–11,18,20,24,28,33,35,42,49,53 with 2D spiral
microchannels used to achieve the desired radius of
curvature and flow path length, a unique aspect of the
present work is that the spiral microchannel is formed
by wrapping a straight microchannel around the cir-
cumference of the syringe body, thus defining the
geometry required to develop Dean vortices and
achieve particle focusing. Previous studies have
explored 3D helical microchannels for inertial particle
separations,41,46 as well as the development of
microfluidic devices for cell separation that can be
manually attached to a syringe48 or pipette.17,38 The
present work extends these advances by directly inte-
grating inertial focusing and plasma collection ele-
ments into a conventional venipuncture syringe. This
approach greatly reduces fabrication complexity, and
eliminates the need for a separate microfluidic device
to perform plasma separation, simplifying the opera-
tional workflow by reducing the number of compo-
nents, fluidic interconnects, and manual steps.
Reducing assay steps is an important consideration for
the design of robust diagnostics,5,27,31 and is consistent
with the ASSURED criteria developed by the World
Health Organization to benchmark the utility of dis-
ease diagnostics for the developing world.30
The integration of the microfluidic separation
component onto the cylindrical surface of a syringe
body is enabled by the use of solvent casting as a
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AND D. L. DEVOE
simple thermoplastic soft lithography technique that is
amenable to high throughput and low cost fabrica-
tion.16 The integrated devices were used to successfully
remove up to 99.7% of blood cells at the syringe outlet.
Significantly, efficient separations were achieved using
manual thumb actuation of the syringe plunger, sug-
gesting that the reported technology may be of par-
ticular value for application to diagnostics in highly
resource-constrained settings.
MATERIALS AND METHODS
Materials
Thermoplastic cyclic olefin polymer (COP; Zeonor
1020R) pellets were obtained from Zeon Chemicals
(Louisville, KY). Polydimethylsiloxane (PDMS; Syl-
gard 184) was purchased from Dow Corning Corpo-
ration (Auburn, MI). Negative photoresists (SU-8
2100) was purchased from MicroChem (Westborough,
MA). Ethanol (‡ 99.5%), phosphate buffered saline
(PBS, tablet), and bovine serum albumin (BSA, pow-
der) were purchased from Sigma-Aldrich (St. Louis,
MO). Decahydronaphthalene (decalin; cis + trans,
98%) was purchased from Alfa Aesar (Haverhill, MA).
Polymethylmethacrylate (PMMA) sheets, 4.5 mm
thick, were purchased from US Plastics (Lima, OH).
Viton O-rings with 4.8 mm outer diameter and 1 mm
cross section were obtained from McMaster Carr
(Elmhurst, IL). COP syringes (1 mL volume) were
purchased from Merit Medical OEM (Salt Lake City,
UT). The body of each COP syringe was approxi-
mately 8 cm long, with an outer diameter of 1.0 cm.
Blood samples were procured from healthy volunteers
using an approved protocol through the University of
Maryland Health Center.
Thermoplastic Patterning by Solvent Casting
Thin COP films with molded microfluidic features
were fabricated by thermoplastic solvent casting,
which enables the fabrication of pliable solvent-laden
thermoplastic substrates containing high resolution
microchannels and other microscale features.16 First,
the desired microfluidic feature was patterned by SU-8
photolithography on a silicon wafer, and its reverse
mold was formed by PDMS soft lithography. After
fully curing the PDMS layer, a solution of solvated
COP was prepared by mixing COP pellets with decalin
(27:73 w/w) and poured over the PDMS mold before
covering the solution with a second piece of unpat-
terned PDMS. Decalin was gradually removed from
the COP solution through the bulk PDMS by placing
the mold on a custom vacuum curing chuck for 10 h at
 Plasma Isolation in a Syringe by Conformal Integration
room temperature. The curing setup is described in
detail in Supplementary Fig. S1. Once cured, the pat-
terned COP film was peeled from the PDMS mold in
preparation for device bonding. Both films were 9 cm
long and 4 cm wide, resulting in approximately 2.5
wraps around the syringe body at the end of the inte-
gration process. For each device, a 150 lm thick COP
film was patterned with the main separation and
plasma collection channels, and a second 100 lm thick
COP film was patterned with side channels to connect
the separation and plasma collection channels after
device bonding (Fig. 1). The separation channel turns
after traversing the side channel array to position the
terminus over a drilled port in the syringe positioned
behind the plunger tip, allowing the syringe volume
behind the sample to be used for waste collection.
Syringe Integration
The syringe integration process is described in
Fig. 1. First, the surface of the COP syringe body was
polished with 100 and 320 grit sandpaper to remove
the surface coating and yield a smooth surface. An
inlet hole connecting the syringe body to microfluidic
device was drilled in the syringe barrel with a 750 lm
diameter bit. For the DFF devices, waste collection
ports were formed after bonding the microfluidic layer
to the syringe body to prevent misalignment during
assembly. Inlet and plasma collection ports were
opened in the patterned COP films using a 1 mm
diameter hole punch prior to bonding, enabling col-
lection of purified samples at the end of the plasma
collection channel located on the outer surface of the
syringe body. Plasma collection was performed using a
pipette for small volume collection, or by depositing
plasma into a vial through a tygon tube attached to the
channel outlet for continuous collection of larger vol-
umes.
The bonding process was performed by first expos-
ing the outer surface of the COP syringe to a 35% (v/v)
solution of decalin in ethanol for 10 min before
aligning the patterned solvent cast COP films to the
syringe. Each solvent-casted film, which remains pli-
able due to decalin in the bulk polymer, is readily
shaped by hand to conform with the syringe body.
Integration of the microfluidic device was achieved by
first aligning and partially attaching the inlet part of
the main channel layer perpendicular to the syringe
length, then the side channel layer, which also serves as
main channel capping layer, was placed on top of the
main channel layer, and wrapped around the syringe
barrel together. The assembly was placed in the reu-
sable plastic bag, and the air was slowly removed using
a vacuum packaging tool (NutriChef vacuum sealer,
PyleUSA, Brooklyn, NY) to apply uniform radial
pressure and promote intimate contact of the stacked
films and cylindrical syringe surface during bonding.
Bonding was completed by thermally curing the device
in an oven at 60 C for 5 min. After curing, the device
was removed from the vacuum bag and held at room
temperature overnight to ensure evaporation of excess
solvent. For waste collection chamber, a hole was
drilled with a 750 lm diameter bit at the separation
channel terminus after the device is completely cured
to minimize deformation of assembled device. The
open top of the drilled hole is sealed with a transparent
adhesive tape to direct flow to the waste chamber
within the syringe. To seal the waste during syringe
actuation, a circumferential groove was formed in
syringe plunger to house a Viton O-ring, preventing
waste leakage out of the syringe body. Prior to use, the
microchannel surfaces were passivated with 5 mg ml21
BSA in 1 9 PBS solution.
Separation Element Design
For the spiral DFF separation, channel dimensions
were selected using analytic models of Dean flow
focusing performance within a curved microchan-
nel.1,6,29,53 For a syringe with an outer diameter of
1 cm, channel cross sectional dimensions of 100 lm
width and 40 lm height were chosen. A total spiral
path length of 8 cm was employed. The first 6.2 cm
spiral segment serves to develop Dean vortices within
the channel, while the final 1.8 cm long section con-
tains a series of bifurcation points that allow plasma to
be extracted through multiple side channels after
focusing blood cells away from the channel wall, sim-
ilar to an approach reported by White and coworkers
for planar DFF chips.4 The side channels were de-
signed with 50 lm width, 10 lm height, and 2 mm
length, and were evenly spaced by 2 mm to maintain
approximately 100:1 flow rate ratio between the main
channel and side channel at each bifurcation point to
minimize disturbance to the Dean flow focusing. All
side channels were connected to a 100 lm high and
200 lm wide collection channel that routes the sepa-
rated plasma to an outlet port, while the main spiral
channel exits through a waste port connected to a
sealed chamber within the syringe body. The devel-
opment of Dean flow was first simulated in Ansys
Fluent 16.1 using a simpler geometry with the same
cross-sectional dimensions and spiral length approxi-
mately 33% of the length used in the fabricated de-
vices. From these simulations, flow rates above 100 lL
min21 were found to be sufficient to induce Dean
vortices within the spiral length of a single loop
(Supplementary Fig. S2).
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 J. Y. HAN AND D. L. DEVOE

FIGURE 1. Fabrication process for COP microfluidic device integration by solvent casting. (a) Solvated COP is poured onto a
negative PDMS mold prepared from an SU-8 master to form the separation and plasma collection channels, then covered with a flat
PDMS sheet on a heated plate for partial curing. A second COP layer is prepared using the same process to form an array of side
channels that will connect the separation channel and plasma collection channel in the final device. (b) After partial curing, each
patterned COP layer is peeled off from the PDMS mold, and (c) the separation channel layer is placed on a syringe body with
alignment for fluidic connections, then covered with a secondary COP film (upside down) containing side channels. Both layers
are then wrapped around the COP syringe barrel approximately 2.5 times to form the desired spiral flow path, followed by solvent
bonding of the assembly in a vacuum sealer. (d) Cut view showing the bonded COP film layers wrapped on a syringe, with the side
channel array bridging the separation channel and plasma collection channel.

RESULTS residual solvent within the molded COP was found to

Device Fabrication
Integration of the microfluidic separation compo-
nents required a unique approach to device fabrication
due to the cylindrical shape of the syringe body. To
this end, we employed a thermoplastic soft lithography
method based on solvent casting.16 The solvent casting
process consists of dissolving polymer in an organic
solvent, pouring the polymer solution over a solvent-
permeable mold, and solvent removal to harden the
molded thermoplastic structure.16,36 COP was selected
as the thermoplastic material because of its favorable
properties including excellent acid/base and organic
solvent compatibility, low water absorption, and high
optical transmittance at both visible light and ultravi-
olet wavelengths, together with the availability of
commercially-available COP syringes to ensure uni-
form material properties for the integrated devices.
Using a viscous solution of COP dissolved in decalin,
pattern transfer from PDMS molds could be achieved
with resolution below 15 lm (Fig. 2). Furthermore,
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promote a robust and permanent bond between the
various COP layers, including the syringe body, similar
to conventional thermoplastic solvent-mediated
bonding processes. Because both the microfluidic lay-
ers and syringe body were manufactured from COP,
no plasticizing or cracking was observed after bonding.
The vacuum chuck used for the solvent casting
process was designed to direct solvent removal through
the bottom side of the mold by taking advantage of the
high solvent permeability of PDMS. Rapid removal of
solvent through the vacuum chuck not only minimized
unwanted distortion of PDMS from accumulating
solvent, but also removed air through the gas-perme-
able PDMS that would otherwise form voids in the
molded COP films, allowing high-fidelity replica
molding. Unidirectional solvent removal also enabled
the cured COP films to retain solvent on the surface
opposite to the vacuumed side, promoting strong sol-
vent-mediated bonding with the mating COP surfaces
during the final device bonding step.
To maintain intimate contact between all mating
interfaces during the final bonding and solvent removal
 Plasma Isolation in a Syringe by Conformal Integration
FIGURE 2. Cross sectional SEM images of a DFF device after
syringe integration. (a) Side channel and (b) separation
channel features. (c) Waste channel showing the bonding
interfaces between the stacked COP films wrapped around the
syringe. The interfaces are marked based on measured
thicknesses of each film, and show seamless bonding
between the layers. Scale bars are 25, 50, and 100 lm,
respectively.
steps, each assembled device was placed in a plastic bag
connected to a vacuum pump. Upon removal of air
from the bag, uniform pressure was applied to the
outer surface of the syringe/microchannel assembly,
generating a consistent normal force over the entire
surface and ensuring conformal bonding of the
microchannel layers to the syringe body. Radial com-
pression caused by the vacuum bag resulted in minimal
deformation of the shallow side channels (Fig. 2a),
with + 5.4 ± 1.0 and 2 0.4 ± 1.2 lm changes to
width and height, respectively. On the other hand,
deformation of the larger separation channels (Fig. 2b)
was measured to be 2 15.0 ± 0.8 lm in height, 2
0.6 ± 0.8 lm at the bottom width, and +
16.6 ± 1.3 lm at the top width, resulting in a trape-
zoidal cross section for the separation channel. The
waste channel, also patterned on the main channel
COP layer, also showed deformations of 2 18.8 ±
1.3 lm (height) and + 8.4 ± 4.7 lm (width). Seam-
less bonding across the stacked COP films was con-
firmed by the lack of distinctive boundaries in the cross
section of fabricated devices, as presented in Fig. 2c.
The trapezoidal transformation of the separation
channel is expected to enhance the particle focusing by
establishing an imbalance in the Dean vortex magni-
tude between the narrow inner surface and wider outer
surface of the spiral channel.10,20,42,45 Radial defor-
mation of the waste channel is expected to have min-
imal impact on performance since the change in
hydraulic resistance relative to the separation and side
channels is negligible.
Removal of excess solvent after bonding was found
to be an essential fabrication step to ensure mechanical
integrity of the multilayer devices. The solvent casted
COP films remain pliable until solvent is removed from
the polymer matrix, with insufficient solvent removal
often resulting in blistering near the microchannel inlet
upon flow actuation due to increased fluid pressure. In
some cases, leakage at the bonding interface was also
observed. Allowing solvent to evaporate from the
bonded devices by performing an initial treatment at
60 C for 5 min followed by overnight curing under a
ventilation hood at room temperature was sufficient to
remove residual decalin, providing a permanent bond
without blistering. Solvent removal at temperatures
higher than approximately 70 C often resulted in the
formation of solvent gas pockets at the bonding
interface. Bonding and solvent removal conditions
must be optimized for a given microchannel design to
ensure bonding without channel deformation or col-
lapse, particularly for smaller channel features.
Device Operation
Operation of the lab-on-a-syringe device is sum-
marized in Fig. 3. A syringe with an integrated
microfluidic separation element is pre-filled with a de-
fined volume of sterile buffer to achieve a desired blood
dilution level. After drawing blood into the syringe, air
within the syringe is removed by tapping the syringe
body while holding it vertically and gently depressing
the plunger to eject bubbles through the needle. Fol-
lowing air removal, the needle is capped, and the syr-
inge is shaken for several seconds to mix the whole
blood with the stored buffer. The volume of buffer and
blood draw volume are selected to achieve a desired
dilution ratio. Separations were performed with a fixed
dilution ratio of 25:1, corresponding to 30 lL blood
draw into a 750 lL buffer volume. After capping the
needle, the plunger is then depressed to drive the dilute
blood sample through the drilled syringe outlet port
and into the mating inlet port of the integrated
microfluidic device. As the plunger continues to be
depressed, the dilute blood travels through the
microfluidic element for plasma separation. Cell-rich
effluent is routed to a waste port in the syringe body
connected to a sealed chamber behind the plunger head
for safe storage, while the separated blood plasma is
routed to a collection port. Flow rates up to
250 lL min21 were tested using a syringe pump to
control plunger actuation, and approximately 150 lL
min21 flow rate was achieved by strong manual thumb
press. No degradation of cell separation performance
was observed in the tested flow rate range, consistent
with numerical predictions for Dean vortex formation.
On-Syringe Separation by Dean Flow Focusing
As fluid flows along the length of a curved
microchannel, the parabolic flow velocity profile
within the channel is disrupted, leading to the forma-
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 J. Y. HAN
FIGURE 3. Operation of a lab-on-a-syringe device. (a) A
syringe, prefilled with diluent buffer, is used to (b) draw
blood from a patient. (c) The syringe is capped and manually
agitated to mix the blood/buffer solution, and (d) the syringe
plunger is depressed to force blood solution through the
interconnected microchannels.
tion of secondary circulatory flows (Dean vortices)
perpendicular to the primary flow direction due to the
contribution of centrifugal force from the curved
geometry. The Dean vortex intensity is characterized
by the dimensionless Dean number (De) and Reynolds
number (Re):
rffiffiffiffiffiffiffiffi
Dh
De ¼ Re ð1Þ
2Rc
qUDh
Re ¼ ð2Þ
l within the plasma effluent, the integrated syringe de-
where Dh is the microchannel hydraulic diameter, Rc is
the channel radius of curvature, q is the fluid density,
U is the average linear flow velocity, and l is the dy-
namic fluid viscosity. Within the approximate range of
100 < De < 101, two counter-rotating vortices
become significant, exerting a net force on particles
within the channel and focusing them into size-de-
pendent equilibrium positions within the channel
cross-sections.6,53
The patterning of a curved flow path with an
appropriate radius of curvature is an essential
requirement for effective DFF separations. Conven-
tional 1 mL syringes possess an outer radius around
5 mm, presenting an adequate radius of curvature to
achieve micrometer-scale particle separation using
DFF. This feature opens the possibility of taking
advantage of the cylindrical geometry of the syringe
for DFF-based plasma isolation. In this concept, a
straight microchannel is wrapped around the outer
circumference of the syringe (Fig. 4a), allowing parti-
cles to be focused at specific radial positions within the
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channel. Implementation of this concept requires
careful design of the microchannel to ensure efficient
separation. The curved channel length required to
achieve inertial focusing (Lf) is given by1,6,53:
plh2
Lf ¼ f ð3Þ
qUmax D2p fL
where h is the height of channel, Umax is the maximum
linear flow velocity, Dp is the particle diameter, fL is the
dimensionless lift coefficient, and f is the Dean focus-
ing coefficient which has been empirically observed to
vary between 0.2 and 1.0.1 Using this expression, the
optimal channel dimensions for blood cell separation
were determined for the 1 mL syringe used in this work
(Supplementary Fig. S3a). Similar analysis performed
for different standard syringe dimensions (Supple-
mentary Fig. S3b) reveals that a suitable range of Dean
numbers to support effective separation can also be
achieved on larger syringes up to 20 mL using the
conformal integration method described here. While
not analyzed in detail for our design, the trapezoidal
cross section of the main channel resulting from the
syringe integration process may further promote the
generation of asymmetric Dean vortices, enhancing
particle focusing over wider range of Re.20,24,45
A set of fabricated DFF syringe devices (Figs. 4b–
4d) were used to evaluate blood separation perfor-
mance. The DFF devices were found to be capable of
processing blood at dilution levels as low as 25 9 .
Plasma isolation test results for the DFF devices are
summarized in Fig. 5. Defining separation efficiency as
the percent reduction in blood cell concentration
sign exhibited an average separation efficiency of
99.7% when operated at a flow rate of 250 lL min21
and greater than 96.8% at flow rates above 150 lL
min21 using an automated syringe pump, yielding a
total volume recovery for the separated plasma of
approximately 10%.
To operate the lab-on-a-syringe device without the
need for an external pump, the syringe plunger may
instead be actuated by orienting the device vertically
and placing a fixed weight on the end of the plunger.
The force required to operate the syringe over the same
range of flow rates used for the controlled pumping
experiments, shown on the secondary axis of Fig. 5b,
was found to scale linearly with flow rate, with a force
of 194 N required to reach the maximum flow rate of
250 lL min21. Alternately, the plunger force can be
applied manually by simple thumb press. Average
maximal thumb flexion force at comfortable position
has been reported as 81 N,50 corresponding to a flow
rate of approximately 105 lL min21 in our devices.
Fatigue-induced degradation of thumb flexion force is
 Plasma Isolation in a Syringe by Conformal Integration

FIGURE 4. (a) Schematic of the spiral microchannel geometry formed by wrapping a straight channel around a cylindrical syringe
barrel. (b) Photograph of a fabricated DFF device. The inlet connects to the syringe chamber near the tip. The waste outlet directs
waste back into the syringe body through a secondary port connected to an isolation chamber behind plunger tip. The waste
chamber is sealed with a Viton O-ring (red band in syringe plunger handle) to securely contain the waste. (c) An 8 cm-long straight
channel formed 2.5 loops around the syringe body. (d) Lab-on-a-syringe device loaded with blood solution. (b, d) scale
bars = 1 cm, and (c) scale bar = 2.5 mm.

another factor that can impact device performance. To
further explore this issue, a group of 3 lab volunteers
was asked to depress the syringe plunger for 1 min
while exerting a consistent force perceived to be either
‘‘weak’’, ‘‘moderate’’, or ‘‘strong’’ while monitoring
flow rate. The resulting force regimes, displayed in
Fig. 5b, indicate that while moderate and strong
thumb presses over this time period are sufficient to
operate the device, some loss in separation efficiency is
observed when compared with automated operation.
DISCUSSION
The direct integration of microfluidic technology
with conventional biomedical tools can offer many
advantages by simplifying multiple steps from sample
collection to analytical result, while also enhancing the
accessibility of clinical assays by minimizing reliance
on external instrumentation and power. The lab-on-a-
syringe concept presented here can benefit a range of
blood-based diagnostics requiring plasma isolation,
with particular potential for improved utility in re-
source-limited settings. The integration process was
demonstrated using a COP thermoplastic solvent
casting technique. By utilizing commercial syringes
manufactured with COP bodies, residual solvent in the
patterned films promotes solvent-mediated bonding at
the mated interface, while allowing flexible deforma-
tion during assembly to achieve seamless integration of
the multi-layered DFF element onto the syringe. A
central concern for the fabrication method was whe-
ther the conformal solvent casting process would result
in unwanted deformation of the enclosed microchan-
nels. While deformation of the small side channels was
found to be minimal, the larger separation channels
experienced deformation resulting in a trapezoidal
shape, with approximately 42% difference between the
opposing parallel channel sidewalls. When using con-
sistent solvent casting conditions and bonding vacuum
pressure, the deformation was found to be repeatable,
and the resulting channel cross-section is expected to
result in moderate enhancement of DFF particle
focusing process. The overall fabrication technique is
straightforward and scalable, with alignment of the
mating layers presenting the most challenging aspect of
the process.
Fabricated devices were characterized using
motorized actuation for precise flow control, together
with manual actuation to evaluate performance in
environments with minimal infrastructure. Device
operation from sample loading to plasma collection
was conducted entirely in-syringe without any sample
transfer, only requiring replacement of the syringe
needle with a sealed cap to direct flow to the DFF
device after mixing whole blood with pre-loaded buf-
fer. Plasma isolation exhibited improved separation
efficiency with increasing flow rate from 50 to 250 lL
min21, confirming that Dean vortices are successfully
formed within this regime as predicted from the ana-
lytic model. Dean flow focuses blood cells away from
the outer wall of the spiral channel, allowing the side
channels located at the outer wall to collect cell-free
plasma from main channel flow. In our devices, the
hydraulic resistance ratio between each side channel
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 J. Y. HAN
FIGURE 5. (a) DFF-based plasma separation of 25 3 diluted
whole blood operated by syringe pump at different defined
flow rates, together with manual thumb actuation. Scale
bars = 100 lm. (b) separation efficiency as a function of flow
rate and applied force, with approximate manual force
regimes (weak, moderate, and strong) overlaid.
and the main separation channel was designed to be
100:1 to minimize disturbance to flow focusing,
resulting in approximately 10% recovery of the sepa-
rated plasma. The hydraulic resistance ratio was con-
servative, and further optimization may allow higher
recovery from the device.
Our devices were operated with 25 9 dilution of
whole blood in buffer. Dilution of whole blood prior to
separation is a common requirement for microfluidic
plasma isolation based on DFF. Dilution is necessi-
tated by the high cell concentrations and non-Newto-
nian fluid behavior associated with whole blood which
serves to disrupt focusing positions,25 with typical
reported dilution levels in planar DFF devices ranging
from 20 to 100.28,34,47 At this dilution level, no sign of
clogging was found within microfluidic device. For the
microfluidic-enabled syringes, when the DFF design
was tested with 10 9 dilution, the separation efficiency
BIOMEDICAL
ENGINEERING
SOCIETY
AND D. L. DEVOE
decreased to 47.1 and 89.9%, at 50 and 100 lL min21
flow rates, respectively. Furthermore, the large number
of cells entering the side channels at this lower dilution
level resulted in clogging, as evidenced by formation of
red bands near bifurcation points and a commensurate
increase in back pressure. The volumetric plasma
recovery rate achieved in the present study was
approximately 10% of the feed volume. Although this
value compares favorably with various spiral inertial
microfluidic designs,28,40 it is lower than other passive
microfluidic plasma separators reported similar
throughput.33,37,38,47 However, because the syringe
design takes advantage of serial extraction points
along the spiral flow path, whereas conventional DFF
designs collect purified flow only at a single bifurcation
point near the end of channel, improved recovery from
the microfluidic syringe technology is possible by fur-
ther optimizing the hydraulic resistance and spacing of
each side channel.
When actuating the syringe by manual thumb press
in the moderate force regime, an average efficiency of
92% was observed, slightly lower than the 97% effi-
ciency achieved when using a syringe pump. This result
underscores the potential of the technology for point-
of-care applications where reducing the dependence on
laboratory infrastructure is beneficial. For all actua-
tion methods, maximum throughput was limited to 250
lL min21 due to failure of the bond interface between
COP film and syringe at higher pressures. Paralleliza-
tion of microfluidic elements within a single device may
serve to improve throughput in future designs by tak-
ing advantage of the large surface area available on the
syringe barrel to support multiple DFF separation
channels.
Conclusion
The lab-on-a-syringe concept is based on a novel
hybrid integration approach in which microfluidic
components are seamlessly combined with commercial
off-the-shelf syringes. By taking advantage of a solvent
casting technique, the integration process allows for
seamless and conformal bonding between the ther-
moplastic microfluidic components and the syringe
body. In the case of plasma isolation by inertial Dean
flow fractionation, the size-based separation device
directly leverages the cylindrical geometry of syringe
itself to achieve efficient plasma isolation from sampled
blood. The resulting hybrid devices thus enable sample
collection and preparation within a single disposable
element, with the syringe itself serving as a fluidic
actuator that may be operated with a simple thumb
press. Operation of the separation process is typically
completed within 5 min including sample loading and
collection, and materials cost for each device is below
 Plasma Isolation in a Syringe by Conformal Integration
$5. Further development of the platform may enable
the inclusion of biomarker detection within the system,
resulting in a fully integrated diagnostic tool combin-
ing sample collection, sample preparation, and ana-
lytical assay in a single disposable device. More
generally, the conformal integration process described
here presents a new path for combining complex
microfluidic devices into conventional clinical tools
with non-planar geometries, with potential applica-
tions beyond the lab-on-a-syringe platform investi-
gated in this work.
ELECTRONIC SUPPLEMENTARY MATERIAL
The online version of this article (https://doi.org/10.
1007/s10439-020-02526-9) contains supplementary
material, which is available to authorized users.
ACKNOWLEDGMENTS
This research was supported by NSF Grants
ECCS1609074 and CMMI1562468. The authors
acknowledge the support of the Maryland NanoCenter
and its FabLab.
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