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Annals of Biomedical Engineering ( 2020) ENGINEERING

https://doi.org/10.1007/s10439-020-02526-9 SOCIETY

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- power delivery. This requirement presents a world-to-
mally integrated onto the cylindrical barrel of a conventional chip interface challenge that is especially acute for
venipuncture syringe, forming a spiral inertial separation systems designed to operate in resource-limited clinical
element supporting the isolation of plasma from diluted
whole blood. The cylindrical shape of the syringe itself serves environments, where maximizing device reliability,
to define the flow path required for inertial separation by minimizing operational complexity, maintaining device
transforming a linear microchannel to a spiral topology. The sterility, and reducing the potential for exposure to
hybrid system enables inertial plasma separation by Dean biological hazards are critical concerns.5,27,30,31
flow focusing within the same syringe used for a patient An alternative approach to interfacing microfluidic
blood draw, with the seamlessly interconnected microfluidic
element operated by automated or manual actuation of the chips with supporting components for fluid handling
syringe plunger. Plasma isolation is achieved without the and power delivery is to directly integrate microfluidic
need for external instrumentation. Device design and fabri- functionality into existing clinical tools. In this way,
cation challenges are discussed, and effective plasma isola- microfluidics can add new capabilities such as sample
tion within the system is demonstrated, with a peak preparation or analytical steps to established instru-
separation efficiency above 97% using 25 9 diluted blood.
mentation without imposing the need for additional
interconnections or specialized infrastructure for de-
Keywords—Dean flow focusing, Inertial separation, Solvent
vice operation. Here we explore the integration of a
casting.
microfluidic sample preparation element into a dis-
posable plastic syringe as one of the most ubiquitous
biomedical devices. Blood sampling by syringe
INTRODUCTION
venipuncture is routinely performed as the first step for
Lab-on-a-chip technology is an important enabler blood-based diagnostics and serological biomarker
for a broad range of diagnostic and clinical platforms. screening.43 For molecular diagnostics such as
The miniaturization and integration of analytical immunoassays, sampled whole blood is typically sep-
functions into a single microfluidic chip represents a arated by centrifugation to isolate plasma as an ana-
powerful concept offering numerous advantages for lytic matrix, thereby eliminating interference imposed
increasing performance, reducing cost and complexity, by blood cells on target analyte detection.32 While
and enhancing accessibility of biomedical assays. centrifugation is a straightforward process, it requires
Microfluidic devices typically consist of discrete com- access to laboratory equipment that may not be readily
ponents that must be interfaced with external instru- available in smaller clinics, field environments, and
mentation and fluidic interconnects for sample other resource-limited settings. A variety of small-scale
introduction from upstream assay stages, sample technologies have been developed to perform cen-
delivery to downstream stages, reagent addition, and trifugation with manual power input,2,3,44,51 but like
conventional contrifuges these platforms add multiple
process steps and components, thereby increasing as-
Address correspondence to Don L. DeVoe, Department of
Mechanical Engineering, University of Maryland, College Park,
say complexity. To overcome the limitations
MD 20742, USA. Electronic mail: ddev@umd.edu of centrifugation, a range of microfluidic techniques

 2020 Biomedical Engineering Society

 J. Y. HAN AND D. L. DEVOE

based on active8,14,15,19,21,23,26 and pas- simple thermoplastic soft lithography technique that is
7,12,13,22,28,33,34,37,39,40,45,52
sive separations have been amenable to high throughput and low cost fabrica-
reported for plasma isolation in planar chips, including tion.16 The integrated devices were used to successfully
the development of power-free microfluidic devices remove up to 99.7% of blood cells at the syringe outlet.
capable of isolating plasma from whole blood using a Significantly, efficient separations were achieved using
combination of vacuum pumping and sedimentation- manual thumb actuation of the syringe plunger, sug-
based cell separation.49 While these microfluidic tech- gesting that the reported technology may be of par-
nologies offers advantages over centrifugation for ticular value for application to diagnostics in highly
effective plasma isolation from small volume blood resource-constrained settings.
samples, they still require the use of separate compo-
nents for sample collection and plasma separation.
Here we investigate the integration of microfluidic MATERIALS AND METHODS
plasma separation elements into standard thermo-
plastic syringes, resulting in a seamlessly-integrated Materials
device combining initial blood sample collection and Thermoplastic cyclic olefin polymer (COP; Zeonor
plasma processing. Passive separations that employ 1020R) pellets were obtained from Zeon Chemicals
hydrodynamically-driven flow fields are particularly (Louisville, KY). Polydimethylsiloxane (PDMS; Syl-
appropriate for syringe integration, since flow control gard 184) was purchased from Dow Corning Corpo-
may be achieved through manual or automated actu- ration (Auburn, MI). Negative photoresists (SU-8
ation of the syringe plunger. Plasma isolation is 2100) was purchased from MicroChem (Westborough,
achieved using Dean flow focusing (DFF), an inertial MA). Ethanol (‡ 99.5%), phosphate buffered saline
focusing method that offers high throughput passive (PBS, tablet), and bovine serum albumin (BSA, pow-
particle separation resulting from the summation of der) were purchased from Sigma-Aldrich (St. Louis,
inertial lift force and Dean drag force during particle MO). Decahydronaphthalene (decalin; cis + trans,
transport in a curved microchannel.6,9,11,18,24,28,33,42,45 98%) was purchased from Alfa Aesar (Haverhill, MA).
While conventional microfluidic DFF devices are Polymethylmethacrylate (PMMA) sheets, 4.5 mm
routinely fabricated in planar thick, were purchased from US Plastics (Lima, OH).
chips,1,6,9–11,18,20,24,28,33,35,42,49,53 with 2D spiral Viton O-rings with 4.8 mm outer diameter and 1 mm
microchannels used to achieve the desired radius of cross section were obtained from McMaster Carr
curvature and flow path length, a unique aspect of the (Elmhurst, IL). COP syringes (1 mL volume) were
present work is that the spiral microchannel is formed purchased from Merit Medical OEM (Salt Lake City,
by wrapping a straight microchannel around the cir- UT). The body of each COP syringe was approxi-
cumference of the syringe body, thus defining the mately 8 cm long, with an outer diameter of 1.0 cm.
geometry required to develop Dean vortices and Blood samples were procured from healthy volunteers
achieve particle focusing. Previous studies have using an approved protocol through the University of
explored 3D helical microchannels for inertial particle Maryland Health Center.
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 Thermoplastic Patterning by Solvent Casting
present work extends these advances by directly inte- Thin COP films with molded microfluidic features
grating inertial focusing and plasma collection ele- were fabricated by thermoplastic solvent casting,
ments into a conventional venipuncture syringe. This which enables the fabrication of pliable solvent-laden
approach greatly reduces fabrication complexity, and thermoplastic substrates containing high resolution
eliminates the need for a separate microfluidic device microchannels and other microscale features.16 First,
to perform plasma separation, simplifying the opera- the desired microfluidic feature was patterned by SU-8
tional workflow by reducing the number of compo- photolithography on a silicon wafer, and its reverse
nents, fluidic interconnects, and manual steps. mold was formed by PDMS soft lithography. After
Reducing assay steps is an important consideration for fully curing the PDMS layer, a solution of solvated
the design of robust diagnostics,5,27,31 and is consistent COP was prepared by mixing COP pellets with decalin
with the ASSURED criteria developed by the World (27:73 w/w) and poured over the PDMS mold before
Health Organization to benchmark the utility of dis- covering the solution with a second piece of unpat-
ease diagnostics for the developing world.30 terned PDMS. Decalin was gradually removed from
The integration of the microfluidic separation the COP solution through the bulk PDMS by placing
component onto the cylindrical surface of a syringe the mold on a custom vacuum curing chuck for 10 h at
body is enabled by the use of solvent casting as a
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 Plasma Isolation in a Syringe by Conformal Integration

room temperature. The curing setup is described in PyleUSA, Brooklyn, NY) to apply uniform radial
detail in Supplementary Fig. S1. Once cured, the pat- pressure and promote intimate contact of the stacked
terned COP film was peeled from the PDMS mold in films and cylindrical syringe surface during bonding.
preparation for device bonding. Both films were 9 cm Bonding was completed by thermally curing the device
long and 4 cm wide, resulting in approximately 2.5 in an oven at 60 C for 5 min. After curing, the device
wraps around the syringe body at the end of the inte- was removed from the vacuum bag and held at room
gration process. For each device, a 150 lm thick COP temperature overnight to ensure evaporation of excess
film was patterned with the main separation and solvent. For waste collection chamber, a hole was
plasma collection channels, and a second 100 lm thick drilled with a 750 lm diameter bit at the separation
COP film was patterned with side channels to connect channel terminus after the device is completely cured
the separation and plasma collection channels after to minimize deformation of assembled device. The
device bonding (Fig. 1). The separation channel turns open top of the drilled hole is sealed with a transparent
after traversing the side channel array to position the adhesive tape to direct flow to the waste chamber
terminus over a drilled port in the syringe positioned within the syringe. To seal the waste during syringe
behind the plunger tip, allowing the syringe volume actuation, a circumferential groove was formed in
behind the sample to be used for waste collection. 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
Syringe Integration
BSA in 1 9 PBS solution.
The syringe integration process is described in
Fig. 1. First, the surface of the COP syringe body was
Separation Element Design
polished with 100 and 320 grit sandpaper to remove
the surface coating and yield a smooth surface. An For the spiral DFF separation, channel dimensions
inlet hole connecting the syringe body to microfluidic were selected using analytic models of Dean flow
device was drilled in the syringe barrel with a 750 lm focusing performance within a curved microchan-
diameter bit. For the DFF devices, waste collection nel.1,6,29,53 For a syringe with an outer diameter of
ports were formed after bonding the microfluidic layer 1 cm, channel cross sectional dimensions of 100 lm
to the syringe body to prevent misalignment during width and 40 lm height were chosen. A total spiral
assembly. Inlet and plasma collection ports were path length of 8 cm was employed. The first 6.2 cm
opened in the patterned COP films using a 1 mm spiral segment serves to develop Dean vortices within
diameter hole punch prior to bonding, enabling col- the channel, while the final 1.8 cm long section con-
lection of purified samples at the end of the plasma tains a series of bifurcation points that allow plasma to
collection channel located on the outer surface of the be extracted through multiple side channels after
syringe body. Plasma collection was performed using a focusing blood cells away from the channel wall, sim-
pipette for small volume collection, or by depositing ilar to an approach reported by White and coworkers
plasma into a vial through a tygon tube attached to the for planar DFF chips.4 The side channels were de-
channel outlet for continuous collection of larger vol- signed with 50 lm width, 10 lm height, and 2 mm
umes. length, and were evenly spaced by 2 mm to maintain
The bonding process was performed by first expos- approximately 100:1 flow rate ratio between the main
ing the outer surface of the COP syringe to a 35% (v/v) channel and side channel at each bifurcation point to
solution of decalin in ethanol for 10 min before minimize disturbance to the Dean flow focusing. All
aligning the patterned solvent cast COP films to the side channels were connected to a 100 lm high and
syringe. Each solvent-casted film, which remains pli- 200 lm wide collection channel that routes the sepa-
able due to decalin in the bulk polymer, is readily rated plasma to an outlet port, while the main spiral
shaped by hand to conform with the syringe body. channel exits through a waste port connected to a
Integration of the microfluidic device was achieved by sealed chamber within the syringe body. The devel-
first aligning and partially attaching the inlet part of opment of Dean flow was first simulated in Ansys
the main channel layer perpendicular to the syringe Fluent 16.1 using a simpler geometry with the same
length, then the side channel layer, which also serves as cross-sectional dimensions and spiral length approxi-
main channel capping layer, was placed on top of the mately 33% of the length used in the fabricated de-
main channel layer, and wrapped around the syringe vices. From these simulations, flow rates above 100 lL
barrel together. The assembly was placed in the reu- min21 were found to be sufficient to induce Dean
sable plastic bag, and the air was slowly removed using vortices within the spiral length of a single loop
a vacuum packaging tool (NutriChef vacuum sealer, (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

promote a robust and permanent bond between the
Device Fabrication various COP layers, including the syringe body, similar
Integration of the microfluidic separation compo- to conventional thermoplastic solvent-mediated
nents required a unique approach to device fabrication bonding processes. Because both the microfluidic lay-
due to the cylindrical shape of the syringe body. To ers and syringe body were manufactured from COP,
this end, we employed a thermoplastic soft lithography no plasticizing or cracking was observed after bonding.
method based on solvent casting.16 The solvent casting The vacuum chuck used for the solvent casting
process consists of dissolving polymer in an organic process was designed to direct solvent removal through
solvent, pouring the polymer solution over a solvent- the bottom side of the mold by taking advantage of the
permeable mold, and solvent removal to harden the high solvent permeability of PDMS. Rapid removal of
molded thermoplastic structure.16,36 COP was selected solvent through the vacuum chuck not only minimized
as the thermoplastic material because of its favorable unwanted distortion of PDMS from accumulating
properties including excellent acid/base and organic solvent, but also removed air through the gas-perme-
solvent compatibility, low water absorption, and high able PDMS that would otherwise form voids in the
optical transmittance at both visible light and ultravi- molded COP films, allowing high-fidelity replica
olet wavelengths, together with the availability of molding. Unidirectional solvent removal also enabled
commercially-available COP syringes to ensure uni- the cured COP films to retain solvent on the surface
form material properties for the integrated devices. opposite to the vacuumed side, promoting strong sol-
Using a viscous solution of COP dissolved in decalin, vent-mediated bonding with the mating COP surfaces
pattern transfer from PDMS molds could be achieved during the final device bonding step.
with resolution below 15 lm (Fig. 2). Furthermore, To maintain intimate contact between all mating
interfaces during the final bonding and solvent removal
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 Plasma Isolation in a Syringe by Conformal Integration

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
FIGURE 2. Cross sectional SEM images of a DFF device after must be optimized for a given microchannel design to
syringe integration. (a) Side channel and (b) separation
channel features. (c) Waste channel showing the bonding ensure bonding without channel deformation or col-
interfaces between the stacked COP films wrapped around the lapse, particularly for smaller channel features.
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, Device Operation
respectively.
Operation of the lab-on-a-syringe device is sum-
steps, each assembled device was placed in a plastic bag marized in Fig. 3. A syringe with an integrated
connected to a vacuum pump. Upon removal of air microfluidic separation element is pre-filled with a de-
from the bag, uniform pressure was applied to the fined volume of sterile buffer to achieve a desired blood
outer surface of the syringe/microchannel assembly, dilution level. After drawing blood into the syringe, air
generating a consistent normal force over the entire within the syringe is removed by tapping the syringe
surface and ensuring conformal bonding of the body while holding it vertically and gently depressing
microchannel layers to the syringe body. Radial com- the plunger to eject bubbles through the needle. Fol-
pression caused by the vacuum bag resulted in minimal lowing air removal, the needle is capped, and the syr-
deformation of the shallow side channels (Fig. 2a), inge is shaken for several seconds to mix the whole
with + 5.4 ± 1.0 and 2 0.4 ± 1.2 lm changes to blood with the stored buffer. The volume of buffer and
width and height, respectively. On the other hand, blood draw volume are selected to achieve a desired
deformation of the larger separation channels (Fig. 2b) dilution ratio. Separations were performed with a fixed
was measured to be 2 15.0 ± 0.8 lm in height, 2 dilution ratio of 25:1, corresponding to 30 lL blood
0.6 ± 0.8 lm at the bottom width, and + draw into a 750 lL buffer volume. After capping the
16.6 ± 1.3 lm at the top width, resulting in a trape- needle, the plunger is then depressed to drive the dilute
zoidal cross section for the separation channel. The blood sample through the drilled syringe outlet port
waste channel, also patterned on the main channel and into the mating inlet port of the integrated
COP layer, also showed deformations of 2 18.8 ± microfluidic device. As the plunger continues to be
1.3 lm (height) and + 8.4 ± 4.7 lm (width). Seam- depressed, the dilute blood travels through the
less bonding across the stacked COP films was con- microfluidic element for plasma separation. Cell-rich
firmed by the lack of distinctive boundaries in the cross effluent is routed to a waste port in the syringe body
section of fabricated devices, as presented in Fig. 2c. connected to a sealed chamber behind the plunger head
The trapezoidal transformation of the separation for safe storage, while the separated blood plasma is
channel is expected to enhance the particle focusing by routed to a collection port. Flow rates up to
establishing an imbalance in the Dean vortex magni- 250 lL min21 were tested using a syringe pump to
tude between the narrow inner surface and wider outer control plunger actuation, and approximately 150 lL
surface of the spiral channel.10,20,42,45 Radial defor- min21 flow rate was achieved by strong manual thumb
mation of the waste channel is expected to have min- press. No degradation of cell separation performance
imal impact on performance since the change in was observed in the tested flow rate range, consistent
hydraulic resistance relative to the separation and side with numerical predictions for Dean vortex formation.
channels is negligible.
Removal of excess solvent after bonding was found
On-Syringe Separation by Dean Flow Focusing
to be an essential fabrication step to ensure mechanical
integrity of the multilayer devices. The solvent casted As fluid flows along the length of a curved
COP films remain pliable until solvent is removed from microchannel, the parabolic flow velocity profile
the polymer matrix, with insufficient solvent removal within the channel is disrupted, leading to the forma-
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 J. Y. HAN AND D. L. DEVOE

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
FIGURE 3. Operation of a lab-on-a-syringe device. (a) A
for different standard syringe dimensions (Supple-
syringe, prefilled with diluent buffer, is used to (b) draw mentary Fig. S3b) reveals that a suitable range of Dean
blood from a patient. (c) The syringe is capped and manually numbers to support effective separation can also be
agitated to mix the blood/buffer solution, and (d) the syringe
plunger is depressed to force blood solution through the
achieved on larger syringes up to 20 mL using the
interconnected microchannels. conformal integration method described here. While
not analyzed in detail for our design, the trapezoidal
tion of secondary circulatory flows (Dean vortices) cross section of the main channel resulting from the
perpendicular to the primary flow direction due to the syringe integration process may further promote the
contribution of centrifugal force from the curved generation of asymmetric Dean vortices, enhancing
geometry. The Dean vortex intensity is characterized particle focusing over wider range of Re.20,24,45
by the dimensionless Dean number (De) and Reynolds A set of fabricated DFF syringe devices (Figs. 4b–
number (Re): 4d) were used to evaluate blood separation perfor-
rffiffiffiffiffiffiffiffi mance. The DFF devices were found to be capable of
Dh processing blood at dilution levels as low as 25 9 .
De ¼ Re ð1Þ
2Rc Plasma isolation test results for the DFF devices are
summarized in Fig. 5. Defining separation efficiency as
qUDh the percent reduction in blood cell concentration
Re ¼ ð2Þ
l within the plasma effluent, the integrated syringe de-
sign exhibited an average separation efficiency of
where Dh is the microchannel hydraulic diameter, Rc is
99.7% when operated at a flow rate of 250 lL min21
the channel radius of curvature, q is the fluid density,
and greater than 96.8% at flow rates above 150 lL
U is the average linear flow velocity, and l is the dy-
min21 using an automated syringe pump, yielding a
namic fluid viscosity. Within the approximate range of
total volume recovery for the separated plasma of
100 < De < 101, two counter-rotating vortices
approximately 10%.
become significant, exerting a net force on particles
To operate the lab-on-a-syringe device without the
within the channel and focusing them into size-de-
need for an external pump, the syringe plunger may
pendent equilibrium positions within the channel
instead be actuated by orienting the device vertically
cross-sections.6,53
and placing a fixed weight on the end of the plunger.
The patterning of a curved flow path with an
The force required to operate the syringe over the same
appropriate radius of curvature is an essential
range of flow rates used for the controlled pumping
requirement for effective DFF separations. Conven-
experiments, shown on the secondary axis of Fig. 5b,
tional 1 mL syringes possess an outer radius around
was found to scale linearly with flow rate, with a force
5 mm, presenting an adequate radius of curvature to
of 194 N required to reach the maximum flow rate of
achieve micrometer-scale particle separation using
250 lL min21. Alternately, the plunger force can be
DFF. This feature opens the possibility of taking
applied manually by simple thumb press. Average
advantage of the cylindrical geometry of the syringe
maximal thumb flexion force at comfortable position
for DFF-based plasma isolation. In this concept, a
has been reported as 81 N,50 corresponding to a flow
straight microchannel is wrapped around the outer
rate of approximately 105 lL min21 in our devices.
circumference of the syringe (Fig. 4a), allowing parti-
Fatigue-induced degradation of thumb flexion force is
cles to be focused at specific radial positions within the
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 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 in unwanted deformation of the enclosed microchan-
further explore this issue, a group of 3 lab volunteers nels. While deformation of the small side channels was
was asked to depress the syringe plunger for 1 min found to be minimal, the larger separation channels
while exerting a consistent force perceived to be either experienced deformation resulting in a trapezoidal
‘‘weak’’, ‘‘moderate’’, or ‘‘strong’’ while monitoring shape, with approximately 42% difference between the
flow rate. The resulting force regimes, displayed in opposing parallel channel sidewalls. When using con-
Fig. 5b, indicate that while moderate and strong sistent solvent casting conditions and bonding vacuum
thumb presses over this time period are sufficient to pressure, the deformation was found to be repeatable,
operate the device, some loss in separation efficiency is and the resulting channel cross-section is expected to
observed when compared with automated operation. result in moderate enhancement of DFF particle
focusing process. The overall fabrication technique is
straightforward and scalable, with alignment of the
DISCUSSION mating layers presenting the most challenging aspect of
the process.
The direct integration of microfluidic technology Fabricated devices were characterized using
with conventional biomedical tools can offer many motorized actuation for precise flow control, together
advantages by simplifying multiple steps from sample with manual actuation to evaluate performance in
collection to analytical result, while also enhancing the environments with minimal infrastructure. Device
accessibility of clinical assays by minimizing reliance operation from sample loading to plasma collection
on external instrumentation and power. The lab-on-a- was conducted entirely in-syringe without any sample
syringe concept presented here can benefit a range of transfer, only requiring replacement of the syringe
blood-based diagnostics requiring plasma isolation, needle with a sealed cap to direct flow to the DFF
with particular potential for improved utility in re- device after mixing whole blood with pre-loaded buf-
source-limited settings. The integration process was fer. Plasma isolation exhibited improved separation
demonstrated using a COP thermoplastic solvent efficiency with increasing flow rate from 50 to 250 lL
casting technique. By utilizing commercial syringes min21, confirming that Dean vortices are successfully
manufactured with COP bodies, residual solvent in the formed within this regime as predicted from the ana-
patterned films promotes solvent-mediated bonding at lytic model. Dean flow focuses blood cells away from
the mated interface, while allowing flexible deforma- the outer wall of the spiral channel, allowing the side
tion during assembly to achieve seamless integration of channels located at the outer wall to collect cell-free
the multi-layered DFF element onto the syringe. A plasma from main channel flow. In our devices, the
central concern for the fabrication method was whe- hydraulic resistance ratio between each side channel
ther the conformal solvent casting process would result
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 J. Y. HAN 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-
FIGURE 5. (a) DFF-based plasma separation of 25 3 diluted
whole blood operated by syringe pump at different defined ing advantage of the large surface area available on the
flow rates, together with manual thumb actuation. Scale syringe barrel to support multiple DFF separation
bars = 100 lm. (b) separation efficiency as a function of flow channels.
rate and applied force, with approximate manual force
regimes (weak, moderate, and strong) overlaid.
Conclusion
and the main separation channel was designed to be
100:1 to minimize disturbance to flow focusing, The lab-on-a-syringe concept is based on a novel
resulting in approximately 10% recovery of the sepa- hybrid integration approach in which microfluidic
rated plasma. The hydraulic resistance ratio was con- components are seamlessly combined with commercial
servative, and further optimization may allow higher off-the-shelf syringes. By taking advantage of a solvent
recovery from the device. casting technique, the integration process allows for
Our devices were operated with 25 9 dilution of seamless and conformal bonding between the ther-
whole blood in buffer. Dilution of whole blood prior to moplastic microfluidic components and the syringe
separation is a common requirement for microfluidic body. In the case of plasma isolation by inertial Dean
plasma isolation based on DFF. Dilution is necessi- flow fractionation, the size-based separation device
tated by the high cell concentrations and non-Newto- directly leverages the cylindrical geometry of syringe
nian fluid behavior associated with whole blood which itself to achieve efficient plasma isolation from sampled
serves to disrupt focusing positions,25 with typical blood. The resulting hybrid devices thus enable sample
reported dilution levels in planar DFF devices ranging collection and preparation within a single disposable
from 20 to 100.28,34,47 At this dilution level, no sign of element, with the syringe itself serving as a fluidic
clogging was found within microfluidic device. For the actuator that may be operated with a simple thumb
microfluidic-enabled syringes, when the DFF design press. Operation of the separation process is typically
was tested with 10 9 dilution, the separation efficiency completed within 5 min including sample loading and
collection, and materials cost for each device is below
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 Plasma Isolation in a Syringe by Conformal Integration

$5. Further development of the platform may enable immunolabeling of cellular subsets using acoustic
the inclusion of biomarker detection within the system, microstreaming. Microsyst. Nanoeng. 4:17085, 2018.
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Guan, G., L. Wu, A. A. Bhagat, Z. Li, P. C. Y. Chen, S.
generally, the conformal integration process described Chao, C. J. Ong, and J. Han. Spiral microchannel with
here presents a new path for combining complex rectangular and trapezoidal cross-sections for size based
particle separation. Sci. Rep. 3:1475, 2013.
microfluidic devices into conventional clinical tools 11
Hou, H. W., R. P. Bhattacharyya, D. T. Hung, and J. Han.
with non-planar geometries, with potential applica- Direct detection and drug-resistance profiling of bac-
tions beyond the lab-on-a-syringe platform investi- teremias using inertial microfluidics. Lab Chip 15:2297–
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Jung, J., and K.-H. H. Han. Lateral-driven continuous
This research was supported by NSF Grants magnetophoretic separation of blood cells. Appl. Phys.
ECCS1609074 and CMMI1562468. The authors Lett. 93:223902, 2008.
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acknowledge the support of the Maryland NanoCenter Kendall, E. L., M. S. Wiederoder, J. Y. Han, A. Sposito, A.
Wilson, and D. L. DeVoe. Soft lithography microfabrica-
and its FabLab. tion of functionalized thermoplastics by solvent casting. J.
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