Download Nanosystems Engineering on Microfluidic Platform: Potential

Nanosystems Engineering
on Microfluidic Platform: Potential Applications
in Healthcare and Energy Domain
Nripen Chanda
Sr. Scientist
Council of Scientific & Industrial Research Organization
CMERI, Durgapur 713209, India
CMERI: MST Labs
The Microsystems Technology Laboratory
was established in the year 2006 at The
Central Mechanical Engineering Research
Institute, Durgapur, West Bengal, India
2
Research focus
• Micro-Nano Scale Process
Technologies
• Micro-Nano Systems and Devices
•
•
•
•
Lab-on-chip (microfluidics)
Nanosensors (microfluidics): DBT
funded
Self cleaning patterned surfaces
Textured solar cell: (AGATHA, IndoEU-FP7)
• Geometry: 100nm – 500µm
– Micro machining (material
removal)
– Micro molding, laser
sintering
– Electro-deposition
– Chemical approach, self
assembly/organization
3
Theme of presentation
Integration of nano-scale materials into new and traditional
micro/macro devices for biomedical and energy applications
Nano
materials
Micro
devices
Energy &
Bio-applications
4
Nanosystems engineering
Nanosystems engineering deals with the design, development and
characterization of materials (sized between 1 to 100 nanometer in
at least one dimension).
Water
molecule
Nanospheres
Nanotubes
Nanoshells
Human cell
A period
Tennis ball
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Features of nanosystems engineering
Interdisciplinarity
Mechanics, electronics, fluidics, biology, and chemistry
Heterogeneity
Semiconductor, ceramic, glass, organic
Multifunctionality
Sensing & drug delivery, memory storage and energy applications,
Integration
Integration of nanotechnologies and smart systems into new and
traditional materials, e.g. textiles, glass, paper
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Design of nanoscale materials
Functional Molecules
1. Peptide
Markers
2. Antibody
Analytes of interest
3. Protein
4. Nucleic acid
Nanoparticle core as carrier
5. Drug
6. Fluorescent marker
7. Charged species
Multifunctional
Nanomaterials
• sense, diagnose, describe and qualify a given situation;
• suggest or implement appropriate actions;
• interact with the user, the environment or another smart systems.
7
Looks of nanoscale materials
Spheres
Rods
Cages
Gold nanoparticles
•
•
•
•
Nano Letters, 2009, 9, 1798
Anal. Chem., 2012, 84(21), 9478-84
Proc. Natl. Acad. Sci. (PNAS) USA, 2012, 109, 12426
Bioconjugate Chem., 2014, 25, 1565
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Properties of nanoscale materials
• Lateral dimension: 1 – 100 nm
Polymeric nanomaterials
• Exhibit novel physical, electronic
and optical properties due to
– Two dimensional quantum confinement
– High surface to volume ratio
• Potential applications in wide
range of devices
–
–
–
–
Sensors and actuators
Photovoltaic devices – solar cells
Fuel cells, energy storage
Imaging agents, drug delivery systems
Micro System Technology,
CSIR-CMERI, Durgapur, India
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Applications of nanoscale materials
Point-of-Care diagnostics
− Small and highly sensitive sensors for biologically
relevant molecules/ions detection
− Biochemical analysis (protein/cell analysis)
Toxin detection device
In-vitro drug delivery applications
− Drug screening
− Improved delivery of poorly water soluble drugs
− Imaging of drug delivery sites using imaging
modalities
Targeted drug delivery
10
Applications of nanoscale materials
Fuel cell applications
− Fuel cells use hydrogen and air as fuels and produce water as by product
− The technology uses a nanomaterial to produce electricity
Schematic of a fuel cell
– Micro System Technology, CSIR-CMERI, Durgapur, India
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Use of nano-materials in cancer cell detection
Biological differences between normal and abnormal endothelium
cells for cancer tissue
Normal tissue
Cancer tissue
♦ Tumor endothelium is structurally irregular, heterogeneous, and leaky (larger endothelial pores)
compared to normal vessels.
♦ Through circulatory system, nanoparticles enter into tumor tissue through endothelial pores
(100 nm to 2 µm), known as Enhanced Retention and Permeability effect (passive targeting).
♦ Tumor vessels have been shown to exhibit receptors (e.g., GRP, EGF) that can serve as
potential targets for targeted nanoparticles (active targeting).
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Tumor Avid Gold Nanoparticle (GNP)
Starch-GNP
• GNP
• Carbohydrate
Lung
GumArabic- GNP
Protein-GNP
Liver
Prostate
● GNP
● Glycoprotein
• GNP
• Peptide/Antibody
Gold nanoconjugate
Biomolecule
Target
GNP-starch, GNP-dextran
Carbohydrate
Lungs, Liver cancers
GNP-gumArabic, GNP-gelatin
Glycoprotein
Liver cancers
GNP-Polyethylene glycol
Polymer
Spleen abnormality
GNP-GRPR
Peptide
Prostate, breast cancer
GNP-EGFR
Antibody, Peptide
Pancreatic and colon cancer
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Tumor Homing Small Biomolecule
Advantages in “Small Bio-molecule” delivery platform : (a) ease in penetration of tumor vascular
endothelium, (b) increased diffusion rate in tissue, (c) rapid blood clearance, and (d) low immunogenicity
Thioctic acid modified
GRPR peptide (SS-BBN)
for prostate and breast
cancers
Thioctic acid modified
EGFR peptide (SS-EGF)
for pancreatic and colon
cancers
14
Interaction with Cancer Cell (In-vitro)
GNPs internalized into cancer cells by endocytic mechanisms by using
PEPTIDE – RECEPTOR interaction.
Nuclear envelop
Mitochondria
GNP
in cytoplasm
Cell membrane
TEM image of PC-3 cells after
treatment with GNP-BBN
Chanda N et. al. Nano Letters, 2009
& Bioconjugate chemistry, 2014
The corresponding
XEDS spectrum
10 µm
Dark field images of PC-3 cells
after treatment with bombesin
conjugated GNP (GNP-BBN)
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Cancer Tissue Imaging (In-vitro)
Colon tissue with and without GNP-EGFR peptide, viewed under dark field microscopy
No back scattered light observed
Scattered light observed due to attached GNP
 In DF imaging, single gold nanoparticle can be imaged at very low concentration of GNP-EGFR peptide treatment.
 Significant contrast of peptide conjugated gold nanoparticles provides new opportunities for targeted delivery and
molecular imaging.
16
Detection of Circulating Tumour Cells
Photoacoustic imaging has potential for the early
diagnosis of CTC to prevent metastasis in humans.
Schematic illustration of photoacoustic imaging
CTCs emerge from the primary site into
blood vessel and spread to other organs
Detection of photoacoustic signals by irradiating AuNPs tagged 15 prostate cancer cells/100µL under flow show
strong photoacoustic signals (Right)
17
Chanda N et. al. Pharmaceutical Research, 2011, 28,279–291.
Use of microfluidic platform
• Sample savings – smaller piping needs smaller volumes of fluids
• Faster analyses – surface area-to-volume (SAV) ratio increases and
thus renders any surface phenomenon to become more dominant
than the volume factor – diffusion, surface tension, and surface
effects dominate
– This can actually lead to faster reactions!
• Integration – combine lots of steps onto a single device
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Microfluidics concept
Microfluidics deals with the science and technology of the fluid behavior on the
fundamental microscopic level to give rise to very powerful techniques in
controlling and measuring chemical reactions and physical and biological
processes on the micro and nanoscale.
Laminar vs Turbulent Flow
Re = ρUdh/ μ
-
μ is kinematic viscosity of the fluid
U is a characteristic velocity of fluid
dH is hydraulic diameter of channel.
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Pressure driven flow and particle migration
• Shear induced inertial lift force
 Parabolic velocity profile of Poiseuille flows
 Particles roll down towards microchannel walls
 Directed away from microchannel center
• Saffman
Fl =1.615 ρ ν
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d
2
u
f
-u
p

f 12
du
dy
duf
sgn 
 dy



- uf is the fluid velocity at the location of mass
centre of the particle,
- up is the particle velocity,
- (duf/dy) is the shear rate,
- d is the particle diameter and
- ρ and µ are the fluid density and viscosity.
20
Pressure driven flow and particle migration
•
Wall induced lift force (FWL)
 Flow field around particles disturbed due to presence of walls
 Wall induced asymmetric wake exerts a lift force on particles
 Directed away from the microchannel wall
•
Segre and Silberberg (1962)
Particles equilibrate around channel periphery
“Tubular pinch effect”
Segre and Silberberg, J. Fluid Mech., 1962
Fl > FWl
FWl > Fl
FWl ~ Fl
Fl
FWl
Equilibrium position achieved
Nanoparticles focusing in microchannel
Carrier
phase
(H) = 0.15 mm, (L) = 25 mm,(W) = 3.3/0.8 mm
Converging
zone
Nanoparticles
laden fluid
Carrier
phase
Diverging
zone
Accumulation of fluorescent
nanoparticles (500 nm) at
fluid-fluid interface
Higher concentrations of
nanoparticles at interfaces
Nanoparticles distribution:
fluorescence intensity vs. distance
along the width of channel.
Arun RK, Chaudhury K, Biswas G, Chanda N*,Chakraborty S*. Lab on a Chip, 2014, 14, 3800
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Nanoparticles focusing in microchannel
 Hydrodynamic focusing of fluorescent nanoparticles (30 nm) in microchannel.
 (a-d) Microscopic fluorescent images at 470 nm wavelength at different flow
ratio (25:25, 25:50, 25:100, 25:250).
 (e-h) Plots of fluorescence intensity vs. distance along the width of channel.
The maximum concentration of nanoparticles was observed at two interfacial
zones with separation width of 100 µm at flow ratio of 25:50.
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Nanoparticles focusing in microchannel
Absence of nanoparticles distribution
200 μm
(b)
(a)
100
200 μm
100
(e)
(c)
(g)
(h)
90
Intensity
Intensity
Intensity
50
(d)
105
100
(f)
80
75
200 μm
60
80
Intensity
200 μm
60
40
75
60
25
20
0
0
175
350
525
Transverse distance (m)
700
0
150
300
450
Transverse distance (m)
600
40
0
150
300
450
Transverse distance (m)
600
45
0
150
300
450
Transverse distance (m)
Hydrodynamic focusing of fluorescent nanoparticles (30 nm) in straight microchannel.
24
600
Underlying physics leading to particle focusing
duP/dt = fD + fG + fM + fS
dxP/dt = uP
Distribution of the streamlines on an
x–y plane passing through the central
axis of a converging–diverging channel
3D simulation results of the distributions of 45 nm
particles in a converging diverging channel at a flow
rate of 25 μL min−1, for both the carrier and the
dispersed phases. The flow was imposed along the x
direction and the particles were initially distributed
axially around the central axis of the channel.
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Controlled reaction in microchannel
AgNO3 + NaBH4
SDS
AgNP (40 nm)
Superimposed microscopic image demonstrating the
interaction of AgNPs and H2O2 throughout the patterned
micro-channel
AgNP + H2O2
Ag2O + H2O
NH4OH
[Ag(NH3)2]+
26
Controlled reaction in microchannel
SEM image of the Ag ion precipitate formed at
the interfaces at different sheath flow rates (a)
25 µL/min, (b) 50 µL/min, (c) 100 µL/min and (d)
250 µL/min for 30 min.
(a) Magnified SEM image of silver ion
precipitate which appears as flower-like
structure. (b) EDS spectrum of Ag ion
precipitates showing the presence of Ag.
The peaks of the Si and O elements are
due to the Si substrate.
27
Gold nanosensor in Lab on a chip: As detection
PDMS Channel
DI Water with Arsenic ions
θ = 45°
Gold nanosensors
AuNS
M
As3+
M
M
M
M
M
M
M
M
M
M M
M
Nanosensor
M
M
M
M
M
M
M
M
M
M
M
M
M
M
M
M
M
M
M
M
M
M
M
M M
M M
M
Agglomerated
nanosensors
Binding of As3+ ions to AuNS
to form BLUE aggregates
28
Flow in paper channel
Dye flow in Y-shaped paper channel
(Flow rate: 2-3 µl/min)
Paper contains microchannels made of an
embedded network of fibers through which
a liquid can pass automatically due to
capillary flow. This self-pumping property
of the paper substrate along with its low
cost and easy availability makes it an
attractive substrate for the present studies.
SEM picture of filter paper
29
Paper based microfluidics for arsenic detection
Gold nanosensor
(Au-TA-TG)
in Petri dish
-
Arsenic
recognition
site
Binds with
arsenic
Au-TA-TG
Thioctic acid Thioguanine
(spacer)
(probe)
As
Paper channel
Nanoparticle
aggregation
Arsenic sample
in Petri dish
Nath P, Arun RK,. Chanda N* “A paper based microfluidic
device for the detection of arsenic using a gold nanosensor”
RSC Adv., 2014, 4, 59558-59561
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Sensitivity of gold nanosensor
As
AuNS
Control
1
Absorbance
0.3
10ppm
2
0.2
1ppm
0.1ppm 0.01ppm 0.001ppm
3
4
λmax= 520 nm
1
5
Appearance of
a new broad
peak at 620 nm
2
0.1
3
4
6
1.
2.
3.
4.
5.
6.
Au-TA-TG solution
Au-TA-TG + 10ppm As3+
Au-TA-TG + 1ppm As3+
Au-TA-TG + 0.1ppm As3+
Au-TA-TG + 0.01ppm As3+
Au-TA-TG + 0.001ppm As3+
5
6
0
300
400
500
600
Wavelength (nm)
700
800
31
Arsenic detection on paper microchannel
10ppm
Gold
nanosensor
Arsenic
solution
1ppm
0.1ppm
0.01ppm
0.001ppm
Control
Blue
precipitate
15mm
3mm
On paper strip
10ppm
1ppm
0.1ppm
0.01ppm
0.001ppm
Control
32
Selectivity of gold nanosensor towards arsenic
2
Au-TA-TG + all ions (10ppm) including As (III)
A620/A520
1.5
1
Au-TAT-G +
10ppm Ca(II)
Au-TA-TG +
10ppm K(I)
Ca(II)
K(I)
Au-TAT-G +
10ppm Mg(II)
Au-TA-TG +
10ppm Na(I)
Au-TA-TG + Au-TA-TG + all ions
10ppm Fe(II) (10ppm) except As (III)
0.5
0
As(III)
Mg(II)
Metal ions
Na(I)
Fe(III)
mixture of
ions except
As(III)
33
Construction of detector module
To power
source
White
LED
LCD Display
Sample
Color
sensor
Microcontroller
Internally silver
coated box
RGB array
Prototype of the heavy metal ion detector
Photodetector
34
Uric acid detection on paper strip
Paper strip
(+)AuNPs
TMB
oxTMB
H 2 O2
oxTMB
Colourless
(Bluish green)
Uric acid
Glass slide
TMB
(+)AuNPs
(colourless)
(Bluish green colour)
Red
TMB
TMB + H2O2
H2 O2
(+)AuNPs
Blue
Oxidized TMB
Uric acid
Uric acid
Colourless
Capillary flow
TMB
35
Paper strip assay for uric acid detection
Increasing uric acid concentrations
0 ppm
33 ppm
50 ppm
83 ppm 100 ppm
Kumar A, Hens A, Arun RK, Chatterjee M, Mahato K, Layek K, Chanda N*
Analyst, 2015, DOI: 10.1039/c4an02333a
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Drug delivery applications using
Lab-on-a-chip technology
Drug delivery means controlling mass transport at micro-scale.
Drug
formulation
Drug delivered
to target cells
Drug taken
by the cells
Different stages of drug delivery
37
Drug delivery applications using
Lab-on-a-chip technology
HSA
Microfluidic research paves the way to create a more in-vivo like cellular
microenvironment in-vitro in which cellvolume-to-extracellular fluid volume is
usually greater than one.
HSA
In-vitro
microenvironment
In-vivo blood capillaries
PLGA-Rh-TG
nanoparticles
PLGA
HSA
Rhodamine
+ TG
TEM
38
Polymeric NPs as drug delivery systems
PLGA-Rhod-TG
nanoparticles
HSA
Transport through
microchannel
No change in physicochemical properties of
the nanoparticles
39
Dual-task paradigm: Therapy and Imaging
Imaging capability
Therapeutic response
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Microfluidic fuel cell: Micro-power source
What is hindering the success of fuel cells
in the portable arena?
PDMS channel with
copper electrodes
Formic acid
O2 (Air)
CO2
DAQ
+
Graphite anode
Paper
channel
Tissue Paper
Graphite cathode
H2SO4
Glass plate
Arun RK, Halder S, Chanda N*, Chakraborty S* Lab on a Chip, 2014, 14, 1661-1664
41
Paper based microfluidic fuel cell
HCOOH
2HCOOH  2CO2 + 4H+ + 4e-
Anode
CO2
Tissue
Paper
Cathode
O2 (Air)
O2 + 4H+ + 4e-  2H2O
H2SO4
10 mm
Engraved channel parameters were as follows: length (l) = 25 mm,
width (w) = 6 mm and height (h) = 0.1 mm.
42
Micro-fuel cell performance: OCP
2.0
1.5
t = 1000 minutes
Fuel required = ~ 1 ml
E
(volt)
t = 3500 minutes
Fuel required = 2.5 ml
1.0
0.5
0.0
1000
2000
t (min)
Systematic experimental plan
Number of strokes (k) of Hb pencil
Area of the graphite electrode
Load variation of a single cell (2-10 ohm)
Fuel cell in series
Ag-NP particles on paper
3000
4000
Polarization and Power Curve
Max. current density was
660 mA-cm-2
Max. power density 32
mW-cm-2
Effect of pencil strokes and its area as graphite electrodes
44
Effects of nanoparticles & series connection
Effect of AgNPs : Potential
increases, > 0.3 volt
Six cells in series connection
45
Conclusions

Design of nanoscale materials and study their
transport properties in microfluidic domain for biomedical
(healthcare, biochemical analysis) and environmental
applications
- Microfluidic platform to demonstrate the drug delivery
efficacy
- Toxin and protein detection
- Biologically important molecules, such as amino acids,
hormones, uric acids etc. detection

Design and development of fluidic devices with applications
in energy systems.
46
Current research projects
1. “Design and development of biocompatible micro/nano scale materials
and study their colloidal properties in microfluidic domain” funded by CSIR
India (OLP 101512).
2. “Design and Development of Gold-Iron Oxide Based Smart Magnetic
Nanosensor for Detection and Separation of Heavy Metal Ions” funded by
Department of Biotechnology, India (GAP 101612).
3. Nano patterning of metallic and polymeric thin films funded by CSIR
India (ESC0112)
Mr. Ravi K Arun
Ms. Peuli Nath
Ms. Puja Mitra
Group members
Ms. Preeti Singh
Ms. Nivedita Priyadarshini
Ms. Eleena Tom
THANK YOU
Ms. Manosree Chatterjee
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