TESTING OF MICROFLUIDIC DETERMINISTIC

LATERAL DISPLACEMENT DEVICES FOR

ISOLATION OF CIRCULATING TUMOR CELLS
Amanda Hittelman
Mentor: William C. Tang, UCI

Introduction
Metastatic cancer
Circulating tumor cells (CTCs)

CTCs analysis versus biopsies of metastatic caner

Rarity of CTCs found in the blood (1 mL)

1-10 CTCs
Millions of WBCs
Billions of RBCs

CellSearch
Only method approved by FDA (US Food and Drug
Agency)

EpCAM on CTCs
Epithelial cell adhesion molecule antigen

Magnetically draws CTCs out from the bloodstream

Drawbacks
CTCs may lack EpCAM
Cleaving CTCs from magnetic bead changes properties
Difficult to keep alive

Label-Free Detection
Methods
Different physical/chemical properties of CTCs vs.
blood cells

Filtration

Based on size and deformability

CTCs 15-25 m in diameter
RBCs 6-8 m
WBCs 12-15 m

Dielectrophoresis
Based on size and morphology

Polydimethylsiloxane (PDMS)
Silicon-based organic polymer
Transparent, inert, non-toxic, cheap
Can determine hardness of PDMS by adding different
amounts of curing agent (1:10)

Can bond to different surfaces to create devices

glass

Zongbin Lius Work:

Rapid isolation of cancer cells using microfluidic deterministic
lateral displacement
structure

Work demonstrates testing of 2 microfluidic chips with a

deterministic lateral displacement (DLD) structure
Triangular/circular shaped posts
Flow chamber 35 mm long, 3.5 mm wide

Used particle sized filtration

Calculated critical
particle size 5-6 m
Parameters: shape,
space, tilt angle,
critical particle size

Objectives
To improve upon Lius work and use a cheap and labelfree alternative to CellSearch

Testing various microfluidic devices with DLD

structures
Triangular posts
Differing in length, type of triangle, rotation, shift angle

Uses of simulation and fabrication (approaches)

Methods - Design
Autocad
Inlet
2 outlets
Flow chamber
Ranging from 2.3-2.6 mm in
length
Ranging from 1.5-2.1 mm in
width

Flow
Chambers

Critical particle size equation by

Karabacak
DC = 2 gH
Liu found to be 5-6 m

Methods -Simulation
COMSOL Multiphysics program
Simulation of mixture of particles and
water
Particles
Density: 1000 kg/m3
7 m and 20 m diameter
(7e-6+13e-6*(random(particleindex)<0.4)) m

10 atm pressure difference from inlet

to outlet

Determination of Isolation
Efficiency
X/Y axis of each particle
Which outlet each particle is found
Size of each particle
Compiled in Excel

Fabrication of Device
Create design mask
Photolithography to create
the molding
Using negative photoresist
(SU-8)
Using silicon wafer

Hard bake PDMS (with

curing agent) on mask

Plasma bond mask with

glass to create device

Running fluids through device

Polystyrene beads
Fluorescent (yellow 20 m , grey 10 m)
10 m diameter beads and 20 m diameter beads

Mix with water

Dyed green

Connect tubes to the inlets, and outlets lead to separate test

tubes

Manually using syringe pump at about 4 ml/min

Pump mixture of water and polystyrene beads through inlet
Under microscope view how the particles move when reaching
the outlet

Results
Design A
500

Design B
500

451

450

411

400

400

350

350

300

300

250

250

200

200

150

150

100
50

44

100

57

50

434

450

47

439

56

0
Left
Right
outlet 20 outlet 20
micron micron
particles particles
Particle Type

Left
Right
outlet 7 outlet 7
micron micron
particles particles

Left outlet Right

Left outlet Right
20 micron outlet 20
7 micron outlet 7
particles micron
particles micron
particles
particles
Particle Type

Results
Design
1
2
3
4
5
6
7
8
9
10
11
12

% of 20-micron particles in
the right outlet
56.4%
54.4%
55.3%
53.9%
52.7%
75.8%
60.8%
63.1%
54.8%
44.2%
38.3%
37.5%

% of 7-micron particles in
the right outlet
47.7%
50.3%
47.4%
49.2%
53.1%
67.2%
65.4%
63.9%
48.6%
39.4%
42.6%
42.1%

% of total number of
particles in the right outlet
48.6%
50.7%
48.2%
49.6%
53.0%
68.08%
64.9%
63.8%
49.2%
39.9%
42.1%
41.6%

Discussion
Ideal outcome
Large percentage of % of 20-micron particles in the right
outlet
Percentages closer to 50% in % of 7-micron particles in
the right outlet

Design
1
2
3
4
5
6
7
8
9
10
11
12

% of 20-micron particles in
the right outlet
56.4%
54.4%
55.3%
53.9%
52.7%
75.8%
60.8%
63.1%
54.8%
44.2%
38.3%
37.5%

% of 7-micron particles in
the right outlet
47.7%
50.3%
47.4%
49.2%
53.1%
67.2%
65.4%
63.9%
48.6%
39.4%
42.6%
42.1%

% of total number of
particles in the right outlet
48.6%
50.7%
48.2%
49.6%
53.0%
68.08%
64.9%
63.8%
49.2%
39.9%
42.1%
41.6%

Discussion
Design 6
Highest percentage in % of 20-micron particles in the right
outlet
Highest percentage for % of 7-micron particles in the right
outlet

Design 3
% of 7-micron particles in the right outlet close to 50%
55% for % of 20-micron particles in the right outlet

Design
1
2
3
4
5
6
7
8
9
10
11
12

% of 20-micron particles in
the right outlet
56.4%
54.4%
55.3%
53.9%
52.7%
75.8%
60.8%
63.1%
54.8%
44.2%
38.3%
37.5%

% of 7-micron particles in
the right outlet
47.7%
50.3%
47.4%
49.2%
53.1%
67.2%
65.4%
63.9%
48.6%
39.4%
42.6%
42.1%

% of total number of
particles in the right outlet
48.6%
50.7%
48.2%
49.6%
53.0%
68.08%
64.9%
63.8%
49.2%
39.9%
42.1%
41.6%

Discussion
20 micron diameter particles are not being deflected as
much as would be ideal
When it is deflected, some 7 micron particles are being
deflected as well

In design 6 and 3, there is a higher isolation efficiency

rate of the 20 micron particles vs. 7 micron particles

In other studies, flow chamber was greater

Liu, flow chamber: 35 mm long, 3.5 mm wide
May have a greater isolation efficiency with a longer
chamber

Discussion
Fabrication of devices were not statistically significant
Micro posts appeared to be deformed/missing
When sending particles through the outlet, appeared to
be moving to one outlet more
Hard to determine which outlet had higher amounts of 20
m based on color

Discussion
Future studies
Photolithography process
Attempt positive photoresist
When mixing PDMS, add more curing agent

Fine tune changes for promising designs

Simulations
Recalculate critical particle size for each design
Run simulations of longer flow chambers
Run simulations in blood material instead of water

Acknowledgements
Thank you to my mentor, Professor William C. Tang of
University of California, Irvine, for providing me the
opportunity to conduct research in his lab and sharing
his knowledge and advice throughout the year to assist
me in my project.

Thank you to my graduate student, Tiffany Lin, for

working with me in the lab this year and assisting me
every step of the way.

Special thanks to Southern California Academy of

Sciences (SCAS) for making this possible and assisting
me in writing my paper.

Literature Cited

Andre A. Adams and Paul I. Okagbare. 2008. Highly Efficient Circulating Tumor Cell
Isolation from Whole Blood and Label-Free Enumeration Using Polymer-Based
Microfluidics with an Integrated Conductivity Sensor. Web.

Arpita Chatterjee, Sathyakumar S. Kuntaegowdanahalli, and Ian Papautsky. 2011.

Inertial microfluidics for continuous separation of cells and particles. Microfluidics,
BioMEMS, and Medical Microsystems IX, 7929.

Chao Jin, Sarah M. McFaul, and Simon P. Duffy. Technologies for label-free
separation of circulating tumor cells: from historical foundations to recent
developments. 2014. 16:59:39.

Igor Cima, Chay Wen Yee, Florina S. Iliescu, and Wai Min Phyo. Label-free
isolation of circulating tumor cells in microfluidic devices: Current research and
perspectives. 2012. Web.

Samuel K. Sia and George M. Whitesides. Microfluidic devices fabricated in

poly(dimethylsiloxane) for biological studies. 2003. Electrophoresis. 23: 3563-3576.

Thomas M. Geislinger and Thomas Franke. Sorting of circulating tumor cells (MV3melanoma) and red blood cells using non-inertial lift. 2013. Web.

Zongbin Liu and Fei Huang. 2013. Rapid isolation of cancer cells using microfluidic
deterministic lateral displacement structure. Web.