SUPPORTING INFORMATION
Rare Cell Isolation and Profiling on a Hybrid Magnetic / Size-Sorting (HMSS) Chip
Jaehoon Chung, David Issadore, Adeeti Ullal, KyungHeon Lee, Ralph Weissleder, Hakho Lee
SUPPLEMENTARY METHODS
1. Design of microfluidic channel
The fluidic system had a triple-layer structure that defined the underpass-gap in the size sorter (5 µm in
height), fluidic channel (height, HC = 50 µm), and herringbone patterns (height, HH = 50 µm). The main
microfluidic channel in the magnetic filter region consisted of 64 parallel mixing channels (width, WC =
380 µm). The dimension of herringbone mixer was chosen according to the previous report1 to
maximize the events of suspended cells contacting the channel bottom (see Fig. S1a for details). Cell
capturing structure was implemented in the size sorter region (Fig. S1b). The capture sites were
designed to trap even small cancer cells (> 5 µm in diameter).
Figure S1. Fluidic structures in HMSS. (a) Design parameters for the herringbone mixer: WC = 380 µm; HC = 50
µm; LH1 = 250 µm; LH2 = 130 µm; WH1 = 150 µm; WH2 = 150 µm; HH = 50 µm). (b) Dimension of capture sites in
size-sorter: gap height, 5 µm; pitch of capture site, 40 µm; diameter of capture site, 15 µm, w = 6 µm. (c)
Layout of fluidic channel layer. A foot print is 35 mm × 36 mm. Magnetic capture region consists of 64 parallel
channels (length, 12.5 mm) on the self-assembled magnet surface. The size capture region consists of 2 Vshaped groups. Each group contains 450 single cell capture sites. (d) Magnified view of the cell-capture region.
2. Sample preparation
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Samples were prepared by following the workflows shown in Fig. S2. For initial device characterization,
we used RBC-lysed human blood to facilitate leukocyte staining (Fig. S2a). For all molecular staining
experiments, we used anti-coagulated human whole blood (Fig. S2b).
Figure S2. Sample preparation steps for device characterization (a) and on-chip molecular analyses (b).
3. Microfluidic simulation
We performed 3-dimensional fluidic simulations (COMSOL Multiphysics ver. 4.3a) to map out the
trajectory of magnetic objects in the magnetic filter region. Figure S3 shows representative examples of
particle trajectories inside a herringbone (up) and a laminar (down) fluidic channels. Indeed, the chaotic
advection moved more particles to the self-assembled magnet, and thereby led to higher capture yield.
2
Figure S3. Microfluidic simulation. Three dimensional trajectories of magnetic objects inside a herringbone
(top) and a simple laminar (bottom) channels were calculated. The self-assembled magnetic filter is located on
the channel bottom. The herringbone patterns were not drawn for visual clarity. The dimension of the bounding
box (not to scale) is 380 µm (W) × 100 µm (H) × 12.6 mm (L). The blue dots represent the initial position of
magnetic objects; the red dots indicate the magnetic capture events.
Supplementary Table 1. Comparison of microfluidic-based CTC isolation chips.
Method
Efficiency
(Purity)
Recovery
Enrichment Cellular analysis
rate
2.5 ml/hr
Fluorescence
Positive
staining
(EpCAM based)
1.2 ml/hr
3
Fluorescence
Positive
staining
(EpCAM based)
1 ml/hr
4
Fluorescence
staining
Positive
(size)
> 100 ml/hr
Fluorescence
staining
Positive
(size)
< 0.7 ml/hr
6
~ 3000
Fluorescence
staining
Positive
(size)
> 50 µl/hr
7
Multiple microposts
> 60 %
> 60 %
Herringbo
ne-PDMS
Chip
~ 92 %
~ 95 %
NA
> 95 %
NA
NA
3D membrane
micro-filter
~ 86%
NA
~ 1000
Capture
structure
array
~ 80%
> 95 %
NA
Flow
focusing
+ Ratchet
~ 97 %
Herringbo
nenanopillar
chip
Filter
Gradated
segregati
on
Immunomagnetic
Dead-end
collection
chamber
DEP
(Dielectro
-phoresis)
DEP-FFF
(field flow
fractionati
on)
Hydrodynamic
Flow rate Ref.
Positive
(EpCAM based)
106
Affinity
Fluorescence
staining
Enrichment
Method (+ / -)
~ 95 %
~ 90 %
NA
Fluorescence
staining
Positive
(size)
2 ml/hr
8
Stacked
magnets
~ 90 %
NA
NA
NA
Positive
(EpCAM based)
10 ml/hr
~ 90 %
NA
NA
NA
Positive
(EpCAM based)
1.2 ml/hr
10
ApoStream
~99 %
(double
enrichment)
> 71 %
16
NA
Positive
(Label free)
1 ml/hr
~ 75 %
70 ~ 90 %
NA
NA
Positive
(Label free)
1.2 ml/hr
12
Pinched flow
fractionation
~ 85 %
> 80 %
1.2 × 104
NA
Positive
(size)
1.2 ml/hr
2
5
9
11
13
3
Inertial &
dean
drag
~ 90 %
~ 80 %
NA
NA
Positive
(size)
60 ml/hr
14
Multistage
MOFF
NA
98.9 %
163
NA
Positive
(size)
7.5 ml/hr
15
99 %
NA
> 40
Fluorescence
staining
Positive
(size)
120 ml/hr
16
~ 96%
96 %
< 50
NA
Positive
(Deformability)
7.5 ml/hr
17
MOFF-DEP
(multi-orifice flow
fractionation)
NA
~ 76 %
162
NA
Positive
(Label free)
7.5 ml/hr
~ 90 %
> 87 %
~ 2000
Fluorescen
ce staining
Hybrid
(Pos. & Neg.)
~ 10 ml/hr
DLD
(determini
stic
lateral
displace
Deformab
ment)
ilitybased
separatio
n
Hybrid
HMSS
(Magnetic
+ Size)
18
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