Lecture 5

Microfluidics and biosensing

Tristan Gilet
Jean-Michel Redouté
 Lab-on-Chip (LoC) 2

Goal: Miniaturize and automate the processes required for biological assays

Microfluidics = peripheral (keyboard & monitor) to chemical and biological worlds

è Potential to turn medicine and pharmaceutics from empirical to information-based

F. K. Balagaddé et al., Science 309, 2005

Lab-on-chip products 3
 Microfluidic
UIDIC APPLICATIONS devices
DEFINITION – Market segmentation 4

• Clinical and Veterinary Diagnostics

Laboratory equipment for clinical and veterinary Clinical &
diagnostics Veterinary
Diagnostics
• Point-of-Care Diagnostics
Micro
Out-of-the-lab diagnostic equipment for near-patient Point of
Reaction -
testing, intensive care, doctor’s offices, home testing Care
Flow
Diagnostics
• Pharmaceutical and Life Science Research Chemistry
Microfluidic devices for drug discovery and screening,
genomics, proteomics, cell analysis…
• Analytical Devices Microfluidic
Microfluidic chips and columns for mass spectrometry, Applications
chromatography and HPLC sample preparation Pharma &
Drug
Life Sciences
• Industrial, Environmental and Agro-Food Testing Delivery
Research
Microfluidic-based tests for quality/process control and
water testing (pesticides, bacteria, etc.). Includes
military, security and forensics applications
• Drug delivery Industrial,
Environmental
Analytical
Microfluidic devices for drug delivery, such as inhalers, Devices
microneedles and implantable micropumps & Agro-Food

• Micro Reaction – Flow Chemistry

Microfluidic devices for micro reaction involved in
research or in pilot-production units
©2017 | www.yole.fr | Status of the Microfluidics Industry

Yole Développement 2017

Status of the Microfluidics Industry
The interest to scale down 5

Microfluidics

What is so different about fluid dynamics in the range 3µm – 3mm ?

 Table of content 6

• Lab-On-Chip

• Microscale fluid dynamics

• Droplet microfluidics and cells in chips

• Biosensing and passive microfluidics (capillary & centrifugal)

• Molecular diffusion and evaporation

• Pros and cons of microfluidics

 Flows in the microscopic realm 7

The macro-world The micro-world

Pressure controller
Valves Pressure gauge
= pump + pressure gauge

Tubes Pump Valves Tubes & channels

è Same ingredients ? Same problems ? Same strategies ?

Credit: http://www.corfumachine.com & Microfluidics Lab @ ULg
Flows in the microscopic realm 8

⇢U H inertia
Reynolds number Re =
µ
=
viscous

U = characteristic flow velocity [m/s]

L = characteristic length scale [m] (e.g. channel diameter)
µ = dynamic viscosity = 10-3 Pa.s for water
ρ = density = 103 kg/m3 for water
“Macro”-fluidics Microfluidics

U ⇠ 0.2m/s, H ⇠ 5cm ) Re ⇠ 10000 U ⇠ 2mm/s, H ⇠ 0.05mm ) Re ⇠ 0.1

Turbulent flow è good mixing Laminar flow è poor mixing

Credit: http://alcheme.tamu.edu
 Microscale fluidics – the electrical analogy 9

Microfluidics Electricity

Rectangular microchannel Electrical path

(length L, width W, height H), L >> W >> H (length L, width W, height H), L >> W >> H
Flow rate Q [m3/s] Current i [c/s]
Pressure difference Δp = pi – po [Pa] Voltage difference ΔV [V]

Hydraulic power P = Q Δp Electrical power P = i ΔV

Resistance R = Δp / Q Resistance R = Δp / Q (Ohm's law)

12µL ⇢L
Poiseuille law R = Pouillet's law R =
W H3 WH
µ [Pa.s] = dynamic viscosity ρ = electrical resistivity
Average speed U = Q / (WH) Current density J = i / (W H)
z ⇣ z⌘
Parabolic flow profile u = 6U 1 Flat flow profile
H H

z
pi H po
u(z)
L
 Microscale fluid flows in suspended MEMS 10

MEMS è Small size è Re << 1 è Navier-Stokes equations linear

è F = bU F = damping force applied on the MEMS [N]

b = damping factor, dependent on geometry [kg/s]

Couette flow Squeezed-film flow (Poiseuille)

W F W
µLW µLW 3
b' F U b' 3
L U L

δ δ
 Flows in the microscopic realm 11

The macro-world The micro-world

Valves Pressure gauge

+ molecular diffusion
+ capillary forces (surface tension)
+ Electro-hydrodynamical phenomena
• Electro-osmosis
• Electrophoresis / di-electrophoresis
Tubes Pump • Electrowetting
è Flow control through electrodes
è New ingredients, new strategies !
 Surface tension 12

Surface tension σ = energy cost per unit area for molecules to be at the interface
~ 0.02 J/m2

Wikipedia

Doug Allan @ Telegraph

σ = 0.02 N/m
è Force per unit length
(@contact line, tangent to liquid interface)

Water strider From Multimedia Fluid Mechanics

 Contact angle 13

Smooth surface è Unique contact angle θ

σ σ
SG = SL + cos ✓ θ
σSG σLS θ σLS σSG
SG SL
) cos ✓ =
Hydrophilic Hydrophobic

S = solid
L = liquid
G = gas

Videos from Multimedia Fluid Mechanics

 Surface tension in microtechnology
Droplet self-alignment 14

Undesired stiction after wet etch Capillary-driven self-assembly

a droplet of liquid is placed between a part and a substrate with

ing wetting patterns, the droplet forms a meniscus between the
ns and aligns the part to the substrate. This is called droplet self-
ment (Fig. 7). The phenomenon is a consequence of the surface energy
droplet: the energy is minimized when the surface area of the droplet
imized i.e. when the patterns are perfectly centered and aligned to
Tanaka
other. The et al., Jpn.
confinement of J.
theAppl. Phys.
droplet 32, the
inside 1993
patterns can be
ed by any of the methods discussed in Subsection 2.4.
Sub-micrometer 2D self-alignment

7 Illustration of the droplet self-alignment principle. a) A droplet forms a meniscus

Syms et al., J. MEMS 12; 2003
n a part and a receptor Sariola,
site. b) The PhD
surfacethesis, Aalto
tension of U. (2012)
the droplet self-aligns the part
 Table of content 15

• Lab-On-Chip

• Microscale fluid dynamics

• Droplet microfluidics and cells in chips

• Biosensing and passive microfluidics (capillary & centrifugal)

• Molecular diffusion and evaporation

• Pros and cons of microfluidics

 Pressure from surface tension 16

σ = 0.02 J/m2 = 0.02 Pa/m-1

è Pressure per unit curvature (Laplace)
n
p0
p0 + p Interface

Ri n = normal vector
R1, R2 = curvature radii

From Multimedia Fluid Mechanics

 Droplet microfluidics 17

Several companies today:

Sphere Fluidics , BioRad , Stilla , Droplet Genomics , LiveDrop

Spin-off from Uliège !

S. van Loo et al., Microfluid Nanofluid (2016)

Pay for the Laplace pressure
è Form droplets = individual micro-reactors
Samples confined by surface tension
Separation + transport è immiscible carrier fluid + microchannels
 Droplets in microfluidic networks 18

è Sorting according to contents, e.g., with pneumatic valves

T. Gilet & S. van Loo, Microfluid Nanofluid (2022)

 BrouzesInstitute,
et al., PNAS 106, 2009
Microfluidics for cell assays
Lexington, MA 02421; and bGenetics Department, Harvard Medical School and cHoward Hughes Medical Boston, MA
19
niversity of Colorado, Boulder, CO, and approved
review March 31, 2009)
The need
based microfluidic technology that enables
ening of single mammalian cells. This inte-
ws forScreening for potential
the encapsulation drugcells
of single candidates
and
ent è aqueous microdroplets (1 pL to 10 nL
on average:
an immiscible carrier oil and enables the
• 13 years
f these reactors at a very high-throughput.
• USscreening
l droplet $800 millions
workflow by conducting
xicity Success
• screen. rate 10%
To perform this screen, we first
iability assay that permits the quantitative
and growth within intact droplets. Next, we
Failure
h viability of usually during
encapsulated clinical
human trials,
monocytic
riodwhere 80% Finally,
of 4 days. of the costs have already
we developed an
beenenabling
t library incurred. the identification of the
during the assay read-out. Using the inte-
ology, we screened a drug library for its
The answer
st U937 of Raindance
cells. Taken together our engineers
droplet (2009)
s modular, robust, uses no moving parts, and
Microfluidic
tential applicationschip that screens
including the cytotoxicity
high-through-
s, combinatorial screening, and facilitating
.
of a drug library on U937 cells

Droplet
ity assay microfluidics
! lab-on-a-chip ! emulsion !
è confinement in picoliter droplets
rofluidic approaches present a paradigm
roviding increased throughputs, reduced
 n the microfluidic environment for over 2 weeks. The microcirculation (Puhl et al., 2003). Similar to a human
was designed to be easily adaptable for in situ optical
cross-section much smaller than the cell dia
liver sinusoid, each unit consisted of !250 tightly packed

Cells and organs on chip

sis. After the desired culture time, cells were imaged via
e contrast and fluorescent microscopy. Staining of the
was readily achieved by flowing through fluorescent
hepatocytes were concentrated into the cell cultu
hepatocytes receiving nutrient flow of approximately 100 pL/s.
20
In order to avoid hepatocyte damage caused by high shear
found that designing barrier channels with a
stress (Park et al., 2005), we designed a microfluidic
es, antibodies, or fixative agents. For cell viability structure to mimic the natural endothelial barrier layer in
dimension approaching 1 mm minimized cell
Petri dish environment è different from in vivo (neighboring cells, biochemical cues)
urements within the microfluidic sinusoid, fluorescent the liver sinusoid (Table I). This consisted of closely spaced
dead staining was performed, and cell counts performed deformation at the interface. These effectively pre
parallel channels with dimensions (width, height, length) of
microscopy. Main challenge = to avoid reversing cell differentiation
2 " 1 " 30 mm (Fig. 1). From the Hagen–Poiseuille relations
loaded cells from blocking the flow through
channels, allowing the cells to be gradually carri
Microfluidics è control to reproduce in vivo conditions
bottomand to manage
of the sinusoid in/out products
unit, resulting in comple

e.g., Liver-on-a-chip
= model of hepatic
Lee et al., Biotech.
microarchitecture Bioeng. 97 (5), 2007

50µm
 Spheroids in droplets 21
ll
Resource
Spheroid = from 100 to 10000 cells that form a spherical tissue-like assembly
OPEN ACCESS

C A B
ll
Droplet microfluidics è allows fabrication in series of highly reproducible spheroids
OPEN ACCESS Resource
è facilitates the study of drug testing and cell-cell interaction
C D

A B

C D
E F Merging and incubation
F G
èJ trapping on chip in droplet arrays

E F
H I

R. Tomasi et al., Cell Reports 31 (2020)

G H I

@ULiège (GIGA cancer & µFL & CORD): CT-chip project

G è investigate metastatic niching with
H spheroids
I
Figure 4. Multiplexed Conditions on Liver Spheroids
 Table of content 22

• Lab-On-Chip

• Microscale fluid dynamics

• Droplet microfluidics and cells in chips

• Biosensing and passive microfluidics (capillary & centrifugal)

• Molecular diffusion and evaporation

• Pros and cons of microfluidics

 Biosensors 23

2 è Transduction of a biochemical signal Biochemistry Research Internationa

Cell cultures
Nucleic acid FET devices

Cells
Human samples: + Nanowire array
blood, urine, body fluids Antibodies

Aptamers Nanoparticles

Food samples
Enzymes

Electrodes

Environmental samples:
air, water, soil, vegetation
Signal Signal
Samples Analytes Bioreceptors Transducers conversion and processing
amplification and display

Figure 1: Schematic diagram showing the components of a biosensor. Reproduced after editing from Grieshaber [12].
S. Patel et al., Biochem. Res. Int. 2016
 e worst recorded Ebola virus outbreak in history has the test zone by the contrast label indicates the presence of

Passive systems for Point-of-Care diagnostics

8000 people and killed more than 5000 people, mostly target analyte in the sample. A control zone also tends to be
rica.62 Although pathogen outbreaks in the developed included on the membrane, and this zone contains antibodies 24
relatively rare and generally minor, there are none- that bind to the reporter antibody. The absorbent pad ensures
curringTechnology
instances of = capillary imbibition
contamination of food with steady wicking of the sample uid along the test strip. Many
in hydrophilic
, Staphylococcus aureus, Listeriaporous medium
monocytogenes and E. variations of this general assay design are possible; common
H7 pathogens. 63,64
The gold standard methods for variations include substitution of antibodies with other bio-
pathogens are culture-based assays that provide good recognition elements, or the use of a competitive assay format
Lateral flow test (dipstick) for immunoassays
and selectivity, but require long incubation times rather than a sandwich assay format.
= most popular PoC diagnostic tool
apid responses to outbreaks.65 An alternative strategy
apid analysis is to target protein, nucleic acid, and www.alfascientific.com
ecule biomarkers that(e.g.,arepregnancy, drugs)
pathogen specic.
ove discussion on health-related targets for diagnos-
o means
ècomprehensive,
sensitive to pore and there
size,is also signicant
r molecular diagnostic tests beyond health care. For
no flow control
public service employees are oen tested for illegal
al drugs, and elite athletes are tested for
è oftensubstances.
ce-enhancing limited toMany simple
heavy metals and
ecules such and robust binary
as pesticides tests
and other toxic pollutants
ant targets in environmental analysis,66 while rapid
g and diagnostic tests are also valuable tools for
oduction and other non-health areas of the biotech-
ctor.67,68 Many of the approaches and challenges
for health-related diagnostics are equally applicable
her sectors, and vice versa.

al ow assays
w assays have been one of the most successful and
able formats since the introduction of lateral ow Fig. 2 Petryayeva
Basic design of a lateral & Russ Algar, RSC Adv. 5,assay
flow immunochromatographic 2015
alyte. The dAb deposition zone can be
Passive « capillary » microfluidics
(Turku, Finland). HumanGervais
CRP-free serum
& Delamarche,
r a recessed microchannel for accommo- human CRP to the desired Lab
CRP Chipconcentration.
9, 2009 25

there using an inkjet. The analyte-dAb Abs were labeled using an Alexa Fluor
= Controlled capillary imbibition in defined geometry (è microfluidics)

Advantages
• Quantitative (more than binary)
• Complex procedures (incl. incubation, mixing) with controlled volume & timing
• Still
illary-driven cheap, possibly
microfluidic disposable,
chip for easy, with limited
effecting immunoassays with need for interfacing
one step. (a) A seriesinstrument
of functional micro
è point-of-care
p for performing immunoassays.diagnostics
(b) The position of and interaction between the analyte, dAbs and cAbs a
(see text for details). The PDMS is planar and patterned with lines of cAbs and antigens for the control li
ple collector, the delay valves, the dAb deposition zones with dAbs, the reaction chamber, the capillary pu
 fluorescent streptavidin streptavidin
dye
Example: Immunoassay with capillary microfluidics
41 Page 4 of 9
26
1 min
(A) Off-chip preparation (B) On-chip procedure
1. Gold-conjugated 2. Capture antibody (cAb)
1. Incubation ofSteps
bead with
(C):microbead cAb
of assay 1. Incubation with sample
biotinylated anti-rabbit IgG (cAb)
2.detection
Mixing analyte with
antibody dAb
(dAb) bound to PMMA
(polyclonal, goat)
bound to analyte (with encapsulated fluorescent dye) Volume Vo
Step Solution
rabbit IgG (analyte)
added (µL) used

1 min 1. Rinsing 30 min BSAgold-conjugated

1% PBS 1 5
streptavidin anti-rabbit IgG (dAb)
(polyclonal, donkey)
2. Sample Analyte + dAb 2 1
PMMA
bead fluorescent Fig. 1 Architecture of the IGSS assay implem
biotinylated anti-rabbit IgG (cAb) 3. Rinsing H2microfluidic
O streptavidin
2 functiona1
dye driven chips and beads
(polyclonal, goat) Biomed Microdevices (2018) 20: off-chip
41
Biomed Microdevices (2018) 20: The
41 steps comprise of: 1. incubation
4. Signal amplification fluorescent
Silver staining
beads with biotinylated3 cAb for 30
2

)(B) On-chip procedure

rabbit IgG (analyte) sample (rabbit IgG in BSA1% PBS) with dAb
On-chip procedure 4. Gold-catalysed5. Rinsing 5.HBead
2O fluorescence 1 8
3. Analyte captured complex analyte/dAb beforemeasured.
loading the res
gold-conjugated
1. Incubation with sample anti-rabbit2.IgG (dAb)amplification
silver
Signal staining (thinusing
metallic
silverfilm)
staining
1. Incubation with sample
(polyclonal, donkey) 2. Signal amplification using silver staining TOTAL (C): Steps of assay 7
2. Mixing analyte with dAb
è Signal amplification
Fig. 1 Architecture of the IGSS assay implemented using capillary- microfluidic chip. b The key steps of the on-chip p
Step
driven microfluidic chips and beads functionalized with receptors. a 1. thestarts to flow
incubation of theinanalyte/dAb
the microfluidic
complexchip
with ow
cA
The off-chip steps comprise of: 1. incubation of streptavidin-coated the microfluidic
tion. When chip,
theand 2. the signal
solution amplificatio
has flown for t
1 min
silverexcess
stainingsolution 1.
occurs inleftRinsing
presence
fluorescent beads with biotinylated cAb for 30 min and 2. mixing of in theofloading
gold nanopar
pad
sample (rabbit IgG in BSA1% PBS) with dAb for 1 min to form the dAb. c Summary of the steps involved in the on-c
complex analyte/dAb before loading the resulting solution to the liquidcleanroom
is allowed totissue and
2. for
flow a the
Sample nexttime
specific solution is
as indic
streptavidin
streptavidin streptavidin
streptavidin The first added liquid is a 1% solution o
biotinylated anti-rabbit IgG (cAb) 3. Rinsing
(polyclonal, goat) is allowed
to flow for 1.5 min (i.e. ~53 n
monitoring the4.advancement of the solu
Signal amplification
starts to flow in the microfluidic chip owingrabbit IgG (analyte)
to capillary ac- beadsblock surfacesand
are located andstops
prevent
thenon-specific
formation ofa
Pham et al.,and
Biomed. 5. Rinsing
dAbs Microdevices 20 (41),
up to the receptor 2018
areas. The ass
 41 Page 4 of 9 Biomed Microdevic
41 Page 8 of 9
Example: Immunoassay with capillary microfluidics
(A) Off-chip preparation (B) On-chip procedure 27
(A) IGSS assay on microfluidic chips (B) IGSS assay on m
1. Incubation of bead with cAb 1. Incubation with sample 2. Signal amplification using silver s

41 41PagePage
4 of49of 9 Biomed
BiomedMicrodevices
Microdevic
30 min
Analyte not captured streptavidin Analyte captured
è (A)
(A) No Off-chip
Off-chip
gold preparation
preparation
catalyst
PMMA
(B)(B) On-chip
On-chip procedure
procedure è Gold catalyst
èbeadNofluorescent
silver staining è Silver staining
1. dye
Incubation
1. Incubation of bead
of bead withwith cAb
cAb 1. 1. streptavidin
Incubation
Incubation with
with sample
sample 2.2.
Signal streptavidin
Signalamplification
amplification usingsilver
using silversta
s
è Strong fluorescence è Weak fluorescence

30 min
streptavidin 30 min
streptavidin
2. Mixing analyte with dAb (C): Steps of assay
PMMAPMMA
Volume Volume
beadbead fluorescent
fluorescent Step
streptavidin Solution streptavidin
streptavidin added (µL) used (nL
streptavidin
dye dye
1 min 1. Rinsing BSA 1% PBS 1 53

2. Sample Analyte + dAb 2 175

Fig. (cAb)
6 Performance comparison of the IGSS assays using: (a) capillary- cut-off value for this IGS
biotinylated anti-rabbit IgG
2. Mixing analyte with dAb (C):
driven microfluidic (C):
Steps
3. Steps
Rinsing
(b) of
of assay
assay H2 O 2 140
2. Mixing analyte
(polyclonal, with
goat) dAb chips and 96-well microtiter plates. For all micro-wells correspond
experiments, the dilution4. of gold-conjugated
Signal amplification dAb was 1:2000.
Silver a 1.
staining different
3 concentration
Volume Volume
246
Detection
Step limit ~ 30 ng/mL
Solution Volume Volume
rabbit IgG (analyte)
Fluorescence micrographs correspond Step Solution
to the attenuation of fluorescence absorbance
added(µL) measurements
(µL) used
used(nL)
(nL
from beads by the stained5.silver added
Rinsing
layer after 7 min loading H2silver
O mixture. 1 on 96-well
performed 88 m
gold-conjugated anti-rabbit IgG 1(dAb)
min profile of
2. Fluorescence 1.
1. the Rinsing IGSS assay on
bead-based
Rinsing BSA BSA 1% PBS
capillary-driven
1% PBS 1
were 1taken 5353min
after 20
1 min
(polyclonal, donkey) microfluidic chips. The fluorescence images were taken TOTALafter 7 min Standard deviations703 of 9
2. Sample Pham et al., Biomed.
Analyte +Microdevices
flowing of silver mixture on the chip. Standard deviations of triplicatesdAb 20 (41),
2 2018 175
represent the cut-off value
 Variant: Absorption and UV-vis spectrometry
T. A. Duncombe et al., Anal. Chem. 93, 2021 28

X Z L
Beer-Lambert law: A = log10
i
= "j cj (z)dz
t j 0

A (λ)= absorbance (sometimes named optical density OD)

λ = light wavelength
𝚽i , 𝚽t = incident, transmitted light flux
𝜀j (λ) = molar attenuation coefficient of species j
cj (z) = concentration of species j at position z
L = opticalpubs.acs.org/ac
length through the sample Article

ontaining amino acid point mutations

33
2Q (see
𝚽 previous work for details).
𝚽 t
seededi in antibiotic-free Mueller−
Sigma-Aldrich) in liquid culture for
0 rpm prior to use. Bacterial cultures
mes in additional media prior to the
f single cells. In Figure 4, for GFP-
ting media contained Mueller−Hinton
namycin. In Figure 5,Lfor ergothionase
g media contained Mueller−Hinton
ioneine, and 1 mM isopropyl-β-D-
10724815001, Sigma-Aldrich).
Figure 2. UV protein quantitation in-droplet of BSA-containing
ISCUSSION è Concentration can be found
droplets. (A)from absorbance
Absorbance spectra measurements
and (B) calibration curve for BSA
on Region for Absorbance. While droplets from 10 to 500 μM at 236 and 280 nm, the UVC and UVB
 Example: Immunoassay with capillary microfluidics 29

Microfabrication:
• Trenches for beads è etch of silicon wafer (DRIE)
evices• (2018) 20: 41
Microchannels è patterned layer of negative photoresist (15µm) + cover with hydrophilic tape P

(A) Microfluidic chip layout (B) Integrated beads

air vent

anti-wetting capillary
loading structures pump bead
pads lanes

flow
resistor

1mm 50µm
Operation:
graph •of the microfabricated
Steps Si microfluidic
1 (binding analyte to dAb) chip
and for IGSS cAb
2 (binding deep, etcheddone
to beads) in Si off-chip.
and 20-μm-wide) where beads funct
with •the Beads
main functional
dispensedcomponents being indicated: a
in bead lanes. receptors have been supplied from a suspension passing fr
rrounded with anti-wetting
• Analytes structures,
and staining flow resistor,
(+ rinsing solutionsBbead whendispensed
in between) the chip surface was pads.
in loading still open, whichflow,
Capillary resulted in th
ry pump,sustained
and air vent. All microfluidic
by capillary structures
pump. Resistor are down
to slow packing of beads
& regulate flowafter
rate.the drying of the bead containing
-μm-high photoresist layer. The chip is 15-mm-long and beads are 6 μm in diameter
• Excess
2 of liquid absorbed on the pads at a given time, before the next reagent is dispensed.
97.5 mm ). b SEM micrograph of a bead lane (15-μm-

Pham et al., Biomed. Microdevices 20 (41), 2018

 Capillary microfluidics – design laws 30

Example: rectangular channel of dimensions L >> W >> H

θ
p0 p1 p0 H
Progressive filling (distance x from inlet)
Q(x)
è Poiseuille flow with resistance proportional to x
x
12µx
p0 p1 = Q(x) µ = dynamic viscosity of the liquid
W H3
dx
Incompressibility è Q(x) = W H
dt
2 cos ✓
Laplace law across the liquid interface p1 = p0
H
s
dx H cos ✓ H cos ✓
è 2x = è x(t) = x20 + (t t0 )
dt 3µ 3µ

Spontaneous imbibition only if θ < π/2.

 View Article On

Centrifugal microfluidics 31

Lab on a Chip
Idea: drive liquid plugs in microchannels and chambers thanks to the centrifugal force

Lab-on-a-disk
èMicro- & nano- fluidic research for chemistry, physics, biology, & bioengineering
... be part of something

bigger!
Chem Soc Rev
www.rsc.org/loc Volume 11 | Number 1 | 7 January 2011 | Pages 1–180

Valve + overflow
Published on 01 November 2010. Downloaded on 08/08/2013 02:23:16.

è metering
= volume selection of the plugs Bio
log
Bio Eng
Phys
ics
y y
istr
em
Ch

Pioneers of Miniaturisa
Congratulations to
Lab on a Chip and Corning Inc are pleas
of the Pioneers of Miniaturisation Prize,
The Prize recognises outstanding achie
contributions to the understanding and
nano-scale science.
Stephen Quake invented Microfluidic L
3.0 Unported Licence.

demonstrated the first multilayered act

valves, and pumping system using PDM
seminal work on Droplet generation (P
From right: Harp Minhas (editor Lab on a Chip),
1st high-density microfluidic chips cont
O. Strohmeier et al., Chem. Soc. Rev. 44, 2015
Stephen Quake (2010 prize winner), Po Ki Yuen (Corning Inc)
with thousands of microfluidic valves a
addressable microfluidic reaction cham
6:34:57 AM.

the founder of Fluidigm and Helicos Bio

Fig. 7 Centrifugal microfluidic unit operations for metering and a

Read 101058

defined volume. The excess is gated into a waste chamber. The m

B.S. Lee et al., Lab Chip 10th
11, 2011
Anniversary: Focus on Korea
ISSN 1473-0197
suitable valves. (b) Different aliquoting concepts.98 (With kind perm
CORNING
PAPER
Cho et al.
Fully integrated lab-on-a-disc for simultaneous analysis of biochemistry and
 Table of content 32

• Lab-On-Chip

• Microscale fluid dynamics

• Droplet microfluidics and cells in chips

• Biosensing and passive microfluidics (capillary & centrifugal)

• Molecular diffusion and evaporation

• Pros and cons of microfluidics

 Molecular diffusion 33

J= Drc (Fick’s law)

x @t c = r · J = Dr2 c (Mass conservation)

p
x⇠ Dt
t=0 t>0 c = molecular concentration [mol/m3]
D = diffusion coefficient [m2/s]
= Result of random (Brownian) motion of molecules J = diffusion flux [mol/m2/s]
• Net flux from high concentration to low concentration t = time [s]
• Efficient to mix things at the microfluidic scales x = average distance travelled by diffusion [m]
Substance Medium Molecular weight [g/mol] D [m2/s]
Tobacco mosaic virus Water 4 x 107 3 x 10-12
Human serum albumin Water 69000 6.1 x 10-11
Inulin Water 5500 1.5 x 10-10
Sucrose Water 342 5.2 x 10-10
Glycine Water 75 10-9
Oxygen Water 32 1.8 x 10-9
Oxygen Air 32 2 x 10-5
Phosphorus (T=1250K) Silicon 30 10-18
Water PDMS 18 10-9
 Evaporation in a microchannel 34

Jd = diffusion flux of water vapor in air [mol/m2/s]

Ji = diffusion flux of water across the interface [mol/m2/s]
Atmosphere
Steady state è Jd = Ji
<latexit sha1_base64="gAPwMG1B9o20ozSd9uM44H5UTpE=">AAACCnicbVDLSsNAFJ34rPUVdelmtAiuSiJa3QgFN+Kqgn1AE8JkOmmHzkzCzEQoIWs3/oobF4q49Qvc+TdO2yxs64ELh3Pu5d57woRRpR3nx1paXlldWy9tlDe3tnd27b39lopTiUkTxyyWnRApwqggTU01I51EEsRDRtrh8Gbstx+JVDQWD3qUEJ+jvqARxUgbKbCPMi+M4F0eZJ7ksJfDazij0DywK07VmQAuErcgFVCgEdjfXi/GKSdCY4aU6rpOov0MSU0xI3nZSxVJEB6iPukaKhAnys8mr+TwxCg9GMXSlNBwov6dyBBXasRD08mRHqh5byz+53VTHV35GRVJqonA00VRyqCO4TgX2KOSYM1GhiAsqbkV4gGSCGuTXtmE4M6/vEhaZ1W3Vr24P6/Ua0UcJXAIjsEpcMElqINb0ABNgMETeAFv4N16tl6tD+tz2rpkFTMHYAbW1y/Mhpmt</latexit>

at given 𝞅
⇢ dxi
<latexit sha1_base64="NswTmKVCPd4Q2vzQXh3ZWTxTMX8=">AAACLXicbVBLSwMxGMz6rPVV9eglWAQvll3R6kUo6EEEoYJ9QHdZstlsG5p9kGTFsuQPefGviOChIl79G6btFrR1IDCZmY/kGy9hVEjTHBoLi0vLK6uFteL6xubWdmlntynilGPSwDGLedtDgjAakYakkpF2wgkKPUZaXv9q5LceCRc0jh7kICFOiLoRDShGUktu6TqzvQDeKjezeQipgpfwGNoBR1gLvVhld2pyG/u+gk/TpJpKUrmlslkxx4DzxMpJGeSou6U3249xGpJIYoaE6FhmIp0McUkxI6pop4IkCPdRl3Q0jVBIhJONt1XwUCs+DGKuTyThWP09kaFQiEHo6WSIZE/MeiPxP6+TyuDCyWiUpJJEePJQkDIoYziqDvqUEyzZQBOEOdV/hbiHdDlSF1zUJVizK8+T5knFqlbO7k/LtWpeRwHsgwNwBCxwDmrgBtRBA2DwDF7BEHwYL8a78Wl8TaILRj6zB/7A+P4BtRWpEw==</latexit>

Across the interface: Ji =

Jd M dt
xi = liquid/air interface position [m] at time t = time [s]
𝜌 = liquid water density [kg/m3]
L M = water molar mass [kg/mol]
x Water
Diffusion of water vapor in the air: Jd = Drc
<latexit sha1_base64="OCuzKbIr3p1eEFLjTzVb9WmCWPQ=">AAACB3icbVDLSsNAFJ3UV62vqEtBBovgxpKIVjdCQRfiqoJ9QBPCZDJph04mYWYilJCdG3/FjQtF3PoL7vwbp20W2nrgwuGce7n3Hj9hVCrL+jZKC4tLyyvl1cra+sbmlrm905ZxKjBp4ZjFousjSRjlpKWoYqSbCIIin5GOP7wa+50HIiSN+b0aJcSNUJ/TkGKktOSZ+5njh/A29zJHRDDI4SU8htfQ4chnCGLPrFo1awI4T+yCVEGBpmd+OUGM04hwhRmSsmdbiXIzJBTFjOQVJ5UkQXiI+qSnKUcRkW42+SOHh1oJYBgLXVzBifp7IkORlKPI150RUgM5643F/7xeqsILN6M8SRXheLooTBlUMRyHAgMqCFZspAnCgupbIR4ggbDS0VV0CPbsy/OkfVKz67Wzu9Nqo17EUQZ74AAcARucgwa4AU3QAhg8gmfwCt6MJ+PFeDc+pq0lo5jZBX9gfP4AD6KXfA==</latexit>

vapour
in air
Ji c(x) = water vapor concentration [mol/m3] at position x [m]
D = diffusion coefficient [m2/s]
xi <latexit sha1_base64="xjMsX8K6YuUpQxK32mpnu7daKP4=">AAACB3icbVDLSsNAFJ34rPUVdSnIYBHqpiSi1Y1QcOOySl/QhDCZTtqhM0mYmRRLyM6Nv+LGhSJu/QV3/o3TNgttPXDhcM693HuPHzMqlWV9G0vLK6tr64WN4ubW9s6uubffklEiMGniiEWi4yNJGA1JU1HFSCcWBHGfkbY/vJn47RERkkZhQ41j4nLUD2lAMVJa8swjXH44hdfQCQTCaeyljuBwlGkxS+8bmWeWrIo1BVwkdk5KIEfdM7+cXoQTTkKFGZKya1uxclMkFMWMZEUnkSRGeIj6pKtpiDiRbjr9I4MnWunBIBK6QgWn6u+JFHEpx9zXnRypgZz3JuJ/XjdRwZWb0jBOFAnxbFGQMKgiOAkF9qggWLGxJggLqm+FeIB0IkpHV9Qh2PMvL5LWWcWuVi7uzku1ah5HARyCY1AGNrgENXAL6qAJMHgEz+AVvBlPxovxbnzMWpeMfOYA/IHx+QN6H5ht</latexit>

pv (x)
Ideal gas law c(x) =
RT
Liquid pv = water vapor pressure [Pa]
water R = 8.31 [Pa.m3/mol/K] = gas constant
T = temperature [K]
dxi MD
<latexit sha1_base64="dC7E8XlzbdaSqWujjsUl1iwtMZ4=">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</latexit>

è = rpv
dt ⇢RT
 Evaporation in a microchannel 35

dxi MD
<latexit sha1_base64="dC7E8XlzbdaSqWujjsUl1iwtMZ4=">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</latexit>

= rpv
dt ⇢RT
Atmosphere
at given 𝞅 xi = liquid/air interface position [m] at time t = time [s]
<latexit sha1_base64="fx0R51qOIuOFppILbmBiYFEs5f0=">AAACB3icbVDLSsNAFJ3UV62vqEtBBovgqiSi1Y1QcOOygn1AE8JkOmmHTibDzKRQQnZu/BU3LhRx6y+482+ctllo64ELh3Pu5d57QsGo0o7zbZVWVtfWN8qbla3tnd09e/+grZJUYtLCCUtkN0SKMMpJS1PNSFdIguKQkU44up36nTGRiib8QU8E8WM04DSiGGkjBfaxCDJPxnCc30BvjKQYUlhICuk8sKtOzZkBLhO3IFVQoBnYX14/wWlMuMYMKdVzHaH9DElNMSN5xUsVEQiP0ID0DOUoJsrPZn/k8NQofRgl0hTXcKb+nshQrNQkDk1njPRQLXpT8T+vl+ro2s8oF6kmHM8XRSmDOoHTUGCfSoI1mxiCsKTmVoiHSCKsTXQVE4K7+PIyaZ/X3Hrt8v6i2qgXcZTBETgBZ8AFV6AB7kATtAAGj+AZvII368l6sd6tj3lrySpmDsEfWJ8/xLiZOg==</latexit>

pv = 'psat M = water molar mass [kg/mol]

D = diffusion coefficient [m2/s]
pv = water vapor pressure [Pa]
𝜌 = liquid water density [kg/m3]
R = 8.31 [Pa.m3/mol/K] = gas constant
L T = temperature [K]
x Water
vapour Right above the interface: pv = psat <latexit sha1_base64="jVE6uO3AzZfzbW8zPN92L8/7w/w=">AAACAHicbZDNSsNAFIVv6l+tf1EXLtwMFsFVSUSrG6HgxmUFawttCJPppB06mYSZSaGEbHwVNy4UcetjuPNtnLZZaOuBgY9z7+XOPUHCmdKO822VVlbX1jfKm5Wt7Z3dPXv/4FHFqSS0RWIey06AFeVM0JZmmtNOIimOAk7bweh2Wm+PqVQsFg96klAvwgPBQkawNpZvHyV+1pMRGufoBhWssM59u+rUnJnQMrgFVKFQ07e/ev2YpBEVmnCsVNd1Eu1lWGpGOM0rvVTRBJMRHtCuQYEjqrxsdkCOTo3TR2EszRMazdzfExmOlJpEgemMsB6qxdrU/K/WTXV47WVMJKmmgswXhSlHOkbTNFCfSUo0nxjARDLzV0SGWGKiTWYVE4K7ePIyPJ7X3Hrt8v6i2qgXcZThGE7gDFy4ggbcQRNaQCCHZ3iFN+vJerHerY95a8kqZg7hj6zPHxQylg4=</latexit>

in air psat = saturation pressure [Pa] = fcn. of temperature

<latexit sha1_base64="jVE6uO3AzZfzbW8zPN92L8/7w/w=">AAACAHicbZDNSsNAFIVv6l+tf1EXLtwMFsFVSUSrG6HgxmUFawttCJPppB06mYSZSaGEbHwVNy4UcetjuPNtnLZZaOuBgY9z7+XOPUHCmdKO822VVlbX1jfKm5Wt7Z3dPXv/4FHFqSS0RWIey06AFeVM0JZmmtNOIimOAk7bweh2Wm+PqVQsFg96klAvwgPBQkawNpZvHyV+1pMRGufoBhWssM59u+rUnJnQMrgFVKFQ07e/ev2YpBEVmnCsVNd1Eu1lWGpGOM0rvVTRBJMRHtCuQYEjqrxsdkCOTo3TR2EszRMazdzfExmOlJpEgemMsB6qxdrU/K/WTXV47WVMJKmmgswXhSlHOkbTNFCfSUo0nxjARDLzV0SGWGKiTWYVE4K7ePIyPJ7X3Hrt8v6i2qgXcZThGE7gDFy4ggbcQRNaQCCHZ3iFN+vJerHerY95a8kqZg7hj6zPHxQylg4=</latexit>

pv = psat
xi At the channel outlet, atmosphere at relative humidity 𝝋: pv = 'psat
<latexit sha1_base64="9+dwDv+q+iQnnkO7CqwqBAeWW0U=">AAACCHicbVDLSsNAFJ3UV62vqEsXDhbBVUlEqxuh4MZlBfuAJoTJdNIOnUyGmUmhhCzd+CtuXCji1k9w5984bbPQ1gMXDufcy733hIJRpR3n2yqtrK6tb5Q3K1vbO7t79v5BWyWpxKSFE5bIbogUYZSTlqaaka6QBMUhI51wdDv1O2MiFU34g54I4sdowGlEMdJGCuxjEWSejOE4hzfQGyMphhQWmkI6D+yqU3NmgMvELUgVFGgG9pfXT3AaE64xQ0r1XEdoP0NSU8xIXvFSRQTCIzQgPUM5ionys9kjOTw1Sh9GiTTFNZypvycyFCs1iUPTGSM9VIveVPzP66U6uvYzykWqCcfzRVHKoE7gNBXYp5JgzSaGICypuRXiIZIIa5NdxYTgLr68TNrnNbdeu7y/qDbqRRxlcAROwBlwwRVogDvQBC2AwSN4Bq/gzXqyXqx362PeWrKKmUPwB9bnDyC8mWQ=</latexit>

<latexit sha1_base64="8m45qVLW1rCza8+BAyFxoaD/L+E=">AAACH3icbVBNS8NAEN34WetX1KOXxSLUgyURrV6EghcPHirYD2hKmWw3dnGzCbubQgn5J178K148KCLe+m/ctjlo64OBx3szzMzzY86UdpyxtbS8srq2Xtgobm5t7+zae/tNFSWS0AaJeCTbPijKmaANzTSn7VhSCH1OW/7TzcRvDalULBIPehTTbgiPggWMgDZSz656AnwOOO6lngzxMMPX+BR7gQSS5poCneGye+oNQcYDdpKld1nPLjkVZwq8SNyclFCOes/+9voRSUIqNOGgVMd1Yt1NQWpGOM2KXqJoDOQJHmnHUAEhVd10+l+Gj43Sx0EkTQmNp+rviRRCpUahbzpD0AM1703E/7xOooOrbspEnGgqyGxRkHCsIzwJC/eZpETzkSFAJDO3YjIAk402kRZNCO78y4ukeVZxq5WL+/NSrZrHUUCH6AiVkYsuUQ3dojpqIIKe0St6Rx/Wi/VmfVpfs9YlK585QH9gjX8AZvSh9A==</latexit>

psat (1 ')
è Gradient of vapor pressure: rpv =
Liquid
L
L = channel length [m], from interface to atmosphere
water
<latexit sha1_base64="EzVrWa5RohJXKpM8DYxp8D6q5Jk=">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</latexit>

dxi M Dpsat (1 ')

è =
dt ⇢RT L
Evaporation in a microchannel 36

<latexit sha1_base64="EzVrWa5RohJXKpM8DYxp8D6q5Jk=">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</latexit>

dxi M Dpsat (1 ')

=
dt ⇢RT L
𝞅 = 50%
Variable Meaning Value Unit
M Water molar mass 0.018 [kg/mol]
D Vapor diffusion coefficient (at 27 . 10-6 [m2/s]
37°C)
L psat Saturation pressure (at 37°C) 6300 [Pa]
Water 𝞅 Relative humidity 50% [-]
vapour
in air 𝜌 Liquid water density 1000 [kg/m3]
R Gas constant 8.31 [Pa.m3/m
xi ol/K]
T Temperature 310 [K]
L Channel length 2 . 10-3 [m]
Liquid <latexit sha1_base64="MOh4oJARjqSCLDW+9PC7Q0fRynk=">AAACMHicbZDLSsNAFIYn3q23qks3g0VwVRMv1aXgQpcKVoWmlMn0pB2cSeLMiVhifCM3PopuFBRx61M4jRW8HRj4+P9zOHP+IJHCoOs+OUPDI6Nj4xOTpanpmdm58vzCiYlTzaHOYxnrs4AZkCKCOgqUcJZoYCqQcBqc7/X900vQRsTRMfYSaCrWiUQoOEMrtcr7voQQr/1QM55lvla0nV+1ChB5/qVg7mvR6eI19Y1QcEHd6sZN4fkqpWrN5K1yxa26RdG/4A2gQgZ12Crf++2Ypwoi5JIZ0/DcBJsZ0yi4hLzkpwYSxs9ZBxoWI6bANLPi4JyuWKVNw1jbFyEt1O8TGVPG9FRgOxXDrvnt9cX/vEaK4U4zE1GSIkT8c1GYSoox7adH20IDR9mzwLgW9q+Ud5nNDm3GJRuC9/vkv3CyXvVq1a2jzcpubRDHBFkiy2SVeGSb7JIDckjqhJNb8kCeyYtz5zw6r87bZ+uQM5hZJD/Kef8AN66qzw==</latexit>

water è dxi
' 0.3 µm/s
dt
è Sufficiently fast to significantly modify the volume and
concentration of microfluidic samples in incubation conditions
 Table of content 37

• Lab-On-Chip

• Microscale fluid dynamics

• Droplet microfluidics and cells in chips

• Biosensing and passive microfluidics (capillary & centrifugal)

• Molecular diffusion and evaporation

• Pros and cons of microfluidics

 Advantages of LoCs 38

Advantages offered by microfluidics

1) Volume of sample and reagents è much smaller
2) Analysis time reduced from hours to seconds
3) Increased surface/volume ratio è Better kinetics (heat transfer, separation speed),
high sensitivity
4) Low power consumption, portability, low fabrication cost (sometimes)
5) Multiplexing, parallelization, automation of complex procedures, flow control
6) Coupled to mature detection technologies

Current challenges and bottlenecks

1) Robustness: sample variability, user, storage, manufacturing, surface properties
2) Complete integration: sample collection, injection and preparation
 Microfluidics: The interest to scale down 39

Microfluidics

Chamber dimension 1 cm 1 mm 100 μm 10 μm

Volume 1 mL 1 μL 1 nL 1 pL
# of molecules (concentration 1 nM) 6.1011 6.108 6.105 600
Diffusion time (s) 105 (=28h) 1000 (=17min) 10 0.1
Unit density (#/cm2) 1 100 10000 106
Information density (# values /s/cm2) 10-5 0.1 1000 107
Capillary (Laplace) pressure (bar) 10-4 10-3 10-2 10-1
 What next? 40

Lab-on-chips make about 20% annual growth

Now several research labs @ ULiege

+ R&D in local companies
è Need for engineers with expertise in microfluidics

If you want to learn more about this field

è MECA0008 – Microfluidics